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PROBE FOR DETECTING HEPATITIS B VIRUS AND USE THEREOF

20210130916 ยท 2021-05-06

    Inventors

    Cpc classification

    International classification

    Abstract

    A probe for detecting hepatitis B virus and a method for detecting an insertion site of hepatitis B virus at high efficiency based on the analysis method of next-generation sequencing using the probe is disclosed. A probe can be provided that is capable of confirming the insertion site of HBV in the human genome with a possibility of developing into liver cancer. In addition, by applying the probe to the analysis method of next-generation sequencing, HBV insertion sites in the human genome can be analyzed at low cost and high efficiency.

    Claims

    1. A probe composition for detecting hepatitis B virus (HBV) consisting of nucleotide sequences of SEQ ID NO: 1 to SEQ ID NO: 215.

    2. The probe composition of claim 1, wherein the probe detects an insertion site of hepatitis B virus in the human genome.

    3. The probe composition of claim 1, wherein the probe detects an insertion site of hepatitis B virus (HBV) using the analysis method of next-generation sequencing.

    4. The probe composition of claim 1, wherein the probe is for detecting hepatitis B virus (HBV) applied to the detection of hepatitis B virus of Koreans.

    5. The probe composition of claim 1, wherein the length of the probe is 1 to 215 base pairs.

    6. A kit for detecting hepatitis B virus (HBV), comprising the probe composition according to claim 1.

    7. A method for detecting hepatitis B virus (HBV), wherein the method is a method for detecting hepatitis B virus (HBV) through next-generation sequencing (NGS), the method comprising hybridizing a target sample with a probe composition for detecting hepatitis B virus (HBV) consisting of nucleotide sequences of SEQ ID NO: 1 to SEQ ID NO: 215 to capture a target gene.

    8. The method of claim 7, wherein the hybridizing is performed at a temperature of 65 C. for 16 hours to 24 hours.

    9. The method of claim 7, wherein the target gene is a hepatitis B virus (HBV) gene of Koreans.

    10. A method for detecting hepatitis B virus (HBV), comprising: (a) hybridizing a target sample comprising a target gene with a probe composition for detecting hepatitis B virus (HBV) consisting of nucleotide sequences of SEQ ID NO: 1 to SEQ ID NO: 215 to capture a target gene and amplifying to create a library for next-generation sequencing analysis; and (b) sequencing analyzing the library to confirm an insertion site of hepatitis B virus (HBV) in the human genome.

    11. The method of claim 10, wherein the hybridizing is performed at a temperature of 65 C. for 16 hours to 24 hours.

    12. The method of claim 10, wherein the target gene is a hepatitis B virus (HBV) gene of Koreans.

    13. A method for providing information for the diagnosis of liver cancer, using the method according to claim 10.

    Description

    BRIEF DESCRIPTION OF THE DRAWINGS

    [0013] The above and other objects, features and advantages of the present invention will become more apparent to those of ordinary skill in the art by describing in detail exemplary embodiments thereof with reference to the accompanying drawings, in which:

    [0014] FIG. 1 schematically illustrates a process of analyzing an HBV insertion site;

    [0015] FIG. 2 and FIG. 3 show results of measuring between libraries using Agilent 4200 Tape Station and D1000 Screen Tape; and

    [0016] FIG. 4 shows results of breakpoint analysis of human chromosomes in tumor tissue.

    DETAILED DESCRIPTION OF EXEMPLARY EMBODIMENTS

    [0017] Hereinafter, the present invention will be described in detail.

    [0018] The present invention may detect an insertion site of hepatitis B virus (HBV) located in the human genome at high efficiency based on next-generation sequencing (NGS). Specifically, in a DNA library constructed from a patient's liver tissue, an HBV sequence may be captured with a probe complementary to the self-constructed HBV. Based on this, HBV and breakpoints of the human genome may be detected (refer to FIG. 1).

    [0019] As used herein, the term probe refers to a nucleic acid fragment corresponding to several bases to several hundred bases for specific binding to DNA or RNA, and afterwards, the presence or absence of specific DNA or RNA may be confirmed by amplification, separation, and detection.

    [0020] The present invention provides a probe for detecting hepatitis B virus (HBV) consisting of nucleotide sequences of SEQ ID NO: 1 to SEQ ID NO: 215.

    [0021] The probe may detect an insertion site of hepatitis B virus in the human genome. More specifically, the probe may detect an insertion site of hepatitis B virus (HBV) using the analysis method of next-generation sequencing.

    [0022] The probe may be applied to the detection of hepatitis B virus of Koreans, and more specifically, it may be applied to the detection of genotype hepatitis C virus.

    [0023] The length of the probe is 120 nucleotides. When the length of the probe is too short or too long, false hybridization increases and the likelihood of a decrease in specificity increases. In the present invention, hybridization efficiency was maximized by optimizing the length of a probe as above.

    [0024] In addition, the probe is based on the complete genome sequences of 8 prototypes of hepatitis B virus (HBV) of Koreans, and by allowing each HBV nucleotide sequence to overlap, it is designed to have almost 100% coverage for hepatitis B virus (HBV) of Koreans.

    [0025] In addition, the present invention provides a composition for detecting hepatitis B virus (HBV), including the probe. The composition may include deoxynucleoside triphosphate (dNTP), heat-resistant polymerase, and a metal ion salt such as magnesium chloride and the like, in addition to the probe.

    [0026] In addition, the present invention provides a kit for detecting hepatitis B virus (HBV), including the composition.

    [0027] The kit may include a barcoding primer in which an adapter suitable for the NGS device to be used is combined with a barcode sequence.

    [0028] In addition, the kit may further include a reagent commonly used in a method for detecting nucleic acid. For example, it may include deoxynucleoside triphosphate (dNTP), heat-resistant polymerase, and a metal ion salt such as magnesium chloride and the like that are required for PCR reaction, and may include dNTP, sequenase, and the like that are required for sequencing. In addition, the kit may take the form of a bottle, a tub, a sachet, an envelope, a tube, an ampoule, and the like, and these may be partially or entirely formed from plastic, glass, paper, foil, wax, and the like. The container may be equipped with a completely or partially removable plug, which is initially part of a container or may be attached to the container by mechanical, adhesive, or other means. The container may be equipped with a stopper that may allow access to the contents by an injection needle. The kit may include an external package, and the external package may include instructions for use of the components.

    [0029] The present invention provides a method for detecting hepatitis B virus (HBV), wherein the method is a method for detecting hepatitis B virus (HBV) through next-generation sequencing (NGS), the method including hybridizing a target sample with a probe for detecting hepatitis B virus (HBV) composed of a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 215 to capture a target gene.

    [0030] As used in, the term hybridization means that complementary single-stranded nucleic acids form double-stranded nucleic acids. The degree of complementarity required for hybridization may vary depending on the hybridization conditions, and in particular, if it can be optimized at temperature, it may be preferably optimized to a temperature described in the protocol that can be specified by the probe manufacturer.

    [0031] As used herein, the term target gene refers to a gene sequence to be detected, and it is hybridized with a probe under hybridization, annealing, or amplification conditions.

    [0032] As used herein, the term target gene is not different from the terms used in the present specification such as target gene, target gene sequence, or target sequence, and these terms are used interchangeably in the present specification.

    [0033] As used herein, a target sample refers to a sample including a gene region to be detected, and it may be collected from at least one selected from the group consisting of tissue, blood, serum, saliva, urine, semen, and body fluid, and specifically, it may be liver tissue derived from a patient.

    [0034] In addition, the present invention provides a method for detecting hepatitis B virus (HBV), including (a) hybridizing a target sample including a target gene with a probe for detecting hepatitis B virus (HBV) composed of a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 215 to capture a target gene and amplifying to create a library; and (b) sequencing the library to map the produced nucleotide sequence in the human and HBV reference sequences for analysis to confirm an insertion site of hepatitis B virus (HBV) in the human genome.

    [0035] The hybridizing may be performed at a temperature of 65 C. for 16 hours to 24 hours.

    [0036] Since it is a temperature and time condition that optimizes the efficiency of probe hybridization, the hybridization efficiency may be lowered when an experiment outside this range is performed.

    [0037] The target gene may be a hepatitis B virus (HBV) gene of Koreans.

    [0038] In addition, the present invention may provide a method for providing information for the diagnosis of liver cancer, using the method.

    [0039] Hereinafter, the present invention will be described in more detail through exemplary embodiments. Objects, features, and advantages of the present invention will be easily understood through the following exemplary embodiments. The present invention is not limited to the exemplary embodiment described herein, and may be embodied in other forms. The exemplary embodiments introduced herein are provided in order to sufficiently convey the spirit of the present invention to those of ordinary skill in the technical field to which the present invention pertains. Therefore, the present invention should not be limited by the following exemplary embodiments.

    EXAMPLES

    Example 1: Preparation of Probe for HBV Detection

    [0040] In order to perform next-generation sequencing analysis for the detection of an HBV insertion site, a probe for HBV capture was prepared based on the following complete genome sequences of 8 representative Korean HBV types. Complementary probes were prepared such that each HBV nucleotide sequence overlapped with each other. The probe was synthesized through the HPLC purification method, and the concentration and purity of the synthesized probe were confirmed using the BioAnalyzer device.

    TABLE-US-00001 TABLE 1 HBV Prototype Target Referencecustom-character startcustom-character endcustom-character KR184660.1 Hepatitis B virus isolate SS_3_22, 1custom-character 3207custom-character complete genomecustom-character JN315779.1 Hepatitis B virus genotype C2, 1custom-character 3215custom-character complete genomecustom-character GQ872211.1 Hepatitis B virus, complete genomecustom-character 1custom-character 3215custom-character D23680.1 Hepatitis B virus (B4-HBVST1) 1custom-character 3194custom-character complete genome sequencecustom-character AY641559.1 Hepatitis B virus isolate He53, 1custom-character 3215custom-character complete genomecustom-character isolate 36Y18HCC, AB014395.1 Hepatitis B virus 1custom-character 3119custom-character genomic DNA, complete sequencecustom-character isolate 22Y04HCC, AB014381.1 Hepatitis B virus 1custom-character 3215custom-character genomic DNA, complete sequencecustom-character DQ683578.1 Hepatitis B virus from South Korea, 1custom-character 3215custom-character complete genomecustom-character

    [0041] (Sequence Information)

    [0042] The probe targets the following 8 viruses.

    [0043] complete genome,AY641559.1 Hepatitis B virus isolate He53 (https://www.ncbi.nlm.nih.gov/nuccore/AY641559.1)

    [0044] complete genome,DQ683578.1 Hepatitis B virus from South Korea (https://www.ncbi.nlm.nih.gov/nuccore/DQ683578.1)

    [0045] complete genome,GQ872211.1 Hepatitis B virus (https://www.ncbi.nlm.nih.gov/nuccore/GQ872210.1)

    [0046] complete genome,JN315779.1 Hepatitis B virus genotype C2 (https://www.ncbi.nlm.nih.gov/nuccore/JN315779)

    [0047] complete genome,KR184660.1 Hepatitis B virus isolate SS_3_22 (https://www.ncbi.nlm.nih.gov/nuccore/KR184660.1)

    [0048] complete sequence, isolate 22Y04HCC,AB014381.1 Hepatitis B virus genomic DNA (https://www.ncbi.nlm.nih.gov/nuccore/3582357)

    [0049] complete sequence, isolate 36Y18HCC,AB014395.1 Hepatitis B virus genomic DNA (https://www.ncbi.nlm.nih.gov/nuccore/3551389)

    [0050] D23680.1 Hepatitis B virus (B4-HBVST1) complete genome sequence (https://www.ncbi.nlm.nih.gov/nuccore/D23680.1)

    [0051] Based on the above contents, it was prepared by Tilling density 1X, Boosting: balanced, probe group size: 25.595 kbp, Total probe: 215. The sequence information of each designed probe was shown in Table 2 below.

    TABLE-US-00002 TABLE2 TargetID ProbeID Sequence SEQIDNO KR184660.1HepatitisB probe_HBV_ CTCCACAACATTCCACCAAGCTCTGCT SEQIDNO:1 virusisolateSS_3_22, 012017_1 AGATCCCAGAGTGAGGGGCCTATATTT completegenome TCCTGCTGGTGGCTCCAGTTCCGGAAC AGTAAACCCTGTTCCGACTATTGTCTC ACCCATATCGTC KR184660.1HepatitisB probe_HBV_ AAGCAGGCCTTCACTTTCTCGCCAACT SEQIDNO:2 virusisolateSS_3_22, 012017_10 TACAAGGCCTTTCTGTGTAAACAATAT completegenome CTGCACCTTTACCCCGTTGCCCGGCAA CGGTCAGGTCTCTGCCAAGTATTTGCT GACGCAACCCCC D23680.1HepatitisB probe_HBV_ TTCCTCACATTCATTTACAGGAGGACA SEQIDNO:3 virus(B4-HBVST1) 012017_100 TTATTAATAGATGTGAACAATATGTGG completegenome GCCCTCTTACAGTTAATGAAAAAAGGA sequence GATTAAAATTAATTATGCCTGCTAGGT TCTATCCTAACC D23680.1HepatitisB probe_HBV_ TTACCAAATATTTGCCATTGGACAAAG SEQIDNO:4 virus(B4-HBVST1) 012017_101 GCATTAAACCATATTATCCTGAACATG completegenome CAGTTAATCATTACTTCAAAACTAGGC sequence ATTATTTACATACTCTGTGGAAGGCGG GCATTCTATATA D23680.1HepatitisB probe_HBV_ AGAGAGAAACTACACGCAGTGCCTCA SEQIDNO:5 virus(B4-HBVST1) 012017_102 TTCTGTGGGTCACCATATTCTTGGGAA completegenome CAAGAGCTACAGCATGGGAGGTTGGT sequence CTTCCAAACCTCGACAAGGCATGGGGA CGAATCTTTCTGTT D23680.1HepatitisB probe_HBV_ CCCAATCCTCTGGGATTCTTTCCCGATC SEQIDNO:6 virus(B4-HBVST1) 012017_103 ACCAGTTGGACCCTGCATTCGGAGCCA completegenome ACTCAAACAATCCAGATTGGGACTTCA sequence ACCCCAACAAGGATCATTGGCCAGAG GCAAATCAGGTA D23680.1HepatitisB probe_HBV_ GGAGCGGGAGCATTCGGGCCAGGGTT SEQIDNO:7 virus(B4-HBVST1) 012017_104 CACCCCACCACACGGCGGTCTTTTGGG completegenome GTGGAGCCCGCAGGCTCAGGGCATATT sequence GACAACCGTGCCAGTAGCACCTCCTCC TGCCTCCACCAAT AY641559.1HepatitisB probe_HBV_ CTCCACCACATTCCACCAAGCTCTACT SEQIDNO:8 virusisolateHe53, 012017_105 AGATCCCAGAGTGAGGGGCCTATATTT completegenome TCCTGCTGGTGGCTCCAGTTCCGGAAC AGTAAACCCTGTTCCGACTACTGCCTC ACCCATATCGTC AY641559.1HepatitisB probe_HBV_ AATCTTCTCGAGGACTGGGGACCCTGC SEQIDNO:9 virusisolateHe53, 012017_106 ACCGAACATGGAGAGCACAACATCAG completegenome GATTCCTAGGACCCCTGCTCGTGTTAC AGGCGGGGTTTTTCTTGTTGACAAGAA TCCTCACAATACC AY641559.1HepatitisB probe_HBV_ ACAGAGTCTAGACTCGTGGTGGACTTC SEQIDNO:10 virusisolateHe53, 012017_107 TCTCAATTTTCTAGGGGGAGCACCCAC completegenome GTGTCCTGGCCAAAATTCGCAGTCCCC AACCTCCAATCACTCACCAACCTCTTG TCCTCCAATTTG AY641559.1HepatitisB probe_HBV_ TCCTGGCTATCGCTGGATGTGTCTGCG SEQIDNO:11 virusisolateHe53, 012017_108 GCGTTTTATCATATTCCTCTTCATCCTG completegenome CTGCTATGCCTCATCTTCTTGTTGGTTC TTCTGGACTACCAAGGTATGTTGCCCG TTTGTCCTCT AY641559.1HepatitisB probe_HBV_ ACTTCCAGGAACATCAACTACCAGCAC SEQIDNO:12 virusisolateHe53, 012017_109 GGGACCATGCAAGACCTGCACGATTCC completegenome TGCTCAAGGAACCTCTATGTTTCCCTCT TGTTGCTGTACAAAACCTTCGGACGGA AATTGCACTTG KR184660.1HepatitisB probe_HBV_ ACTGGATGGGGCTTGGCCATAGGCCAT SEQIDNO:13 virusisolateSS_3_22, 012017_11 CGGCGCATGCGTGGAACCTTTGTGGCT completegenome CCTCTGCCGATCCATACTGCGGAACTC CTAGCAGCTTGTTTTGCTCGCAGCCGG TCTGGAGCGAAA AY641559.1HepatitisB probe_HBV_ TATTCCCATCCCATCATCCTGGGCTTTC SEQIDNO:14 virusisolateHe53, 012017_110 GCAAAATTCCTATGGGAGTGGGCCTCA completegenome GTCCGTTTCTCCTGGCTCAATTTACTAG TGCCATTTGTTCAGTGGTTCGCAGGGC TTTCCCCCAC AY641559.1HepatitisB probe_HBV_ TGTTTGGCTTTCAGTTATATGGATGAT SEQIDNO:15 virusisolateHe53, 012017_111 GTGGTATTGGGGGCCAAGTCTGTACAA completegenome CATCTTGAGGCCCTTTATACCTCTATTA CCAATTTTCTTGTGTCTTTGGGTATACA TTTGAACCCT AY641559.1HepatitisB probe_HBV_ AATAAAACCAAACGTTGGGGCTACTCC SEQIDNO:16 virusisolateHe53, 012017_112 CTTAACTTCATGGGATATGTAATTGGA completegenome AGTTGGGGTACTTTACCACAGGAACAT ATTGTACAAAAAATTAAGCAATGTTTT CGGAAACTGCCT AY641559.1HepatitisB probe_HBV_ GTCAATAGACCTATTGATTGGAAAGTA SEQIDNO:17 virusisolateHe53, 012017_113 TGTCAAAGAATTGTAGGTCTTTTGGGA completegenome TTTGCTGCCCCTTTTACACAATGTGGCT ATCCTGCTTTGATGCCTTTATATGCATG TATACAAGCT AY641559.1HepatitisB probe_HBV_ AAGCAGGCTTTCACTTTCTCGTCAACT SEQIDNO:18 virusisolateHe53, 012017_114 TACAAGGCCTTTCTGTGTAAACAATAT completegenome CTGCACCTTTACCCCGTTGCCCGGCAA CGGTCAGGTCTCTGCCAAGTGTTTGCT GACGCAACCCCC AY641559.1HepatitisB probe_HBV_ ACTGGATGGGGCTTGGCCATAGGCCAT SEQIDNO:19 virusisolateHe53, 012017_115 CGGCGCATGCGTGGAACCTTTGTGGCT completegenome CCTCTGCCGATCCATACTGCGGAACTC CTAGCAGCTTGTTTTGCTCGCAGCCGG TCTGGAGCAAAC AY641559.1HepatitisB probe_HBV_ CTTATCGGGACTGACAACTCTGTTGTC SEQIDNO:20 virusisolateHe53, 012017_116 CTCTCTCGGAAATACACCTCCTTCCCA completegenome TGGCTGCTCGGGTGTGCTGCCAACTGG ATCCTGCGCGGGACGTCCTTTGTCTAC GTCCCGTCGGCG AY641559.1HepatitisB probe_HBV_ CTGAATCCCGCGGACGACCCGTCTCGG SEQIDNO:21 virusisolateHe53, 012017_117 GGCCGTTTGGGCCTCTACCGTCCCCTT completegenome CTTCATCTGCCGTTCCGGCCGACCACG GGGCGCACCTCTCTTTACGCGGTCTCC CCGTCTGTGCCT AY641559.1HepatitisB probe_HBV_ TCTCATCTGCCGGTCCGTGTGCACTTC SEQIDNO:22 virusisolateHe53, 012017_118 GCTTCACCTCTGCACGTCGCATGGAAA completegenome CCACCGTGAACGCCCATCCGGTCTTGC CCAAGGTCTTATATAAGAGGACTCTTG GACTCTCAGCAA AY641559.1HepatitisB probe_HBV_ TGTCAACGACCGACCTTGAGGCATACT SEQIDNO:23 virusisolateHe53, 012017_119 TCAAAGACTGTTTGTTTAAAGACTGGG completegenome AGGAGTTGGGGGAGGAGAATAGGTTA ATGATCTTTGTACTAGGAGGCTGTAGG CATAAATTGGTCT KR184660.1HepatitisB probe_HBV_ CTCATCGGGACTGACAACTCGGTTGTT SEQIDNO:24 virusisolateSS_3_22, 012017_12 CTCTCTCGGAAATACACCTCATTCCCA completegenome TGGCTGCTCGGGTGTGCTGCCAACTGG ATCCTGCGCGGGACGTCCTTTGTTTAC GTCCCGTCGGCG AY641559.1HepatitisB probe_HBV_ GTTCACCAGCACCATGCAACTTTTTCA SEQIDNO:25 virusisolateHe53, 012017_120 CCTCTGCCTAATCATCTCTTGTTCATGT completegenome CCTACTGTTCAAGCCTCCAAGCTGTGC CTTGGGTGGCTTTAGGACATGGACATT GACCCGTATAA AY641559.1HepatitisB probe_HBV_ AGAATTTGGAGCTTCTGTGGAGTTGCT SEQIDNO:26 virusisolateHe53, 012017_121 CTCTTTTTTGCCTTCTGACTTCTTTCCTT completegenome CTATTCGAGATCTCCTCGACACCGCCT CTGCTCTCTATCGGGAGGCCTTAGAGT CTCCGGAACA AY641559.1HepatitisB probe_HBV_ TTGTTCACCTCACCATACAGCACTCAG SEQIDNO:27 virusisolateHe53, 012017_122 GCAAGCTATTCTGTGTTGGGGTGAGTT completegenome GATGAACCTGGCCACCTGGGTGGGAA GTAATTTGGAAGATCCTGCATCCAGGG AATTAGTAGTCAG AY641559.1HepatitisB probe_HBV_ CTATGTCAATGTTAATATGGGCCTAAA SEQIDNO:28 virusisolateHe53, 012017_123 ACTCAGACAAATATTGTGGTTTCACAT completegenome TTCCTGTCTTACTTTTGGAAGAGAAAC CGTTCTTGAGTATTTGGTGTCTTTTGGA GTGTGGATTCG AY641559.1HepatitisB probe_HBV_ CACTCCTACCGCTTACAGACCACCAAA SEQIDNO:29 virusisolateHe53, 012017_124 TGCCCCTATCTTATCAACACTTCCGGA completegenome AACTACTGTTGTTAGACGACGAGGCAG GACCCCTAGAAGAAGAACTCCCTCGCC TCGCAGACGAAG AY641559.1HepatitisB probe_HBV_ ATCTCAATCGCCGCGTCGCAGAAGATC SEQIDNO:30 virusisolateHe53, 012017_125 TCAATCTCGGGAATCTCAATGTTAGTA completegenome TCCCCTGGACTCACAAGGTGGGAAATT TTACTGGGCTTTACTCGTCTACTGTACC TATCTTTAATC AY641559.1HepatitisB probe_HBV_ CTGATTGGCAAACTCCCTCCTTTCCTA SEQIDNO:31 virusisolateHe53, 012017_126 ACATTCATTTACAGGAGGACATTATTG completegenome ATAGATGTCAACAATATGTAGGCCCTC TTACAGTTAATGAAAAAAGGAGATTA AAATTAATTATGC AY641559.1HepatitisB probe_HBV_ CTGCTAGGTTTTATCCTAACCTTACCA SEQIDNO:32 virusisolateHe53, 012017_127 AATATTTGCCCTTGGATAAAGGCATTA completegenome AACCTTATTATCCTGAACATGCAGTTA ATCATTACTTCCAAACTAGGCATTATT TACATACTCTGT AY641559.1HepatitisB probe_HBV_ GGAAGGCTGGCATTCTATATAAGAGA SEQIDNO:33 virusisolateHe53, 012017_128 GAAACTACACGCAGCGCTTCATTTTGT completegenome GGGTCACCATATTCTTGGGAACAAGAG CTACAGCATGGGAGGTTGGTCTTCCAA ACCTCGACAAGGC AY641559.1HepatitisB probe_HBV_ ATGGGGACGAATCTTTCTGTTCCCAAT SEQIDNO:34 virusisolateHe53, 012017_129 CCTCTGGGATTCTTTCCCGATCACCAG completegenome TTGGACCCTGCGTTCGGAGCCAACTCA AACAATCCAGATTGGGACTTCAACCCC AACAAGGATCAC KR184660.1HepatitisB probe_HBV_ CTGAATCCCGCGGACGACCCGTCTCGC SEQIDNO:35 virusisolateSS_3_22, 012017_13 GGCCGTTTGGGCCTCTACCGTCCCCTT completegenome CTTCATCTGCCGTTCCGGCCGACCACG GGGCGCACCTCTCTTTACGCGGTCTCC CCGTCTGTGCCT AY641559.1HepatitisB probe_HBV_ TGGCCAGAGGCAAATCAGGTCGGAGT SEQIDNO:36 virusisolateHe53, 012017_130 GGGAGCATTCGGGCCAGGGTTCACCCC completegenome ACCACACGGCGGTCTTTTGGGGTGGAG CCCTCAGGCTCGGGGCATAGTGACACC AGTGCCAGCAGCG isolate probe_HBV_ ACTGGGGACCCTGCACCGAACATGGA SEQIDNO:37 36Y18HCC,AB01439 012017_132 GAACACAACATCAGGATTCCTAGGACC 5.1HepatitisBvirus CCTGCTCGTGTTACAGGCGGGGTTTTT genomicDNA,complete CTTGTTGACAAGAATCCTCACAATACC sequence ACAGAGTCTAGAC isolate probe_HBV_ TCGTGGTGGACTTCTCTCAATTTTCTAG SEQIDNO:38 36Y18HCC,AB01439 012017_133 GGGGAACACCCACGTGTCCTGGCCAA 5.1HepatitisBvirus AATTCGCAGTCCCCAACCTCCAATCAC genomicDNA,complete TCACCAACCTCTTGTCCTCCAATTTGTC sequence CTGGCTATCGC isolate probe_HBV_ TGGATGTGTCTGCGGCGTTTTATCATA SEQIDNO:39 36Y18HCC,AB01439 012017_134 TTCCTCTTCATCCTGCTGCTATGCCTCA 5.1HepatitisBvirus TCTTCTTGTTGGTTCTTCTGGACTACCA genomicDNA,complete AGGTATGTTGCCCGTTTGTCCTCTACTT sequence CCAGGAACA isolate probe_HBV_ TCAACTACCAGCACGGGACCATGCAA SEQIDNO:40 36Y18HCC,AB01439 012017_135 GACCTGCACGATTCCTGCTCAAGGCAC 5.1HepatitisBvirus CTCTATGTTTCCCTCTTGTTGCTGTACA genomicDNA,complete AAACCTTCGGATGGAAACTGCACTTGT sequence ATTCCCATCCCA isolate probe_HBV_ TCATCCTGGGTTTTCGCAAGATTCCTAT SEQIDNO:41 36Y18HCC,AB01439 012017_136 GGGAGTGGGCCTCAGTCCGTTTCTCCT 5.1HepatitisBvirus GGCTCAGTTTACTAGTGCCATTTGTTC genomicDNA,complete AGTGGTTCGTAGGGCTTTCCCCCACTG sequence TTTGGCTTTCA isolate probe_HBV_ GTTATATGGATGATATAGTATTGGGGG SEQIDNO:42 36Y18HCC,AB01439 012017_137 CCAAGTCTGTACAACATCTTGAGTCCC 5.1HepatitisBvirus TTTATACCGCCATTACCAATTTTCTTTT genomicDNA,complete GTCTTTGGGTATACATTTGAACCCTAA sequence TAAAACCAAAC isolate probe_HBV_ GTTGGGGCTACTCCCTGAACTTCATGG SEQIDNO:43 36Y18HCC,AB01439 012017_138 GATATGTAATTGGAAGTTGGGGTACTT 5.1HepatitisBvirus TACCGCAAGACCATATTGTACTAAAAC genomicDNA,complete TCAAGCAATGTTTTCGAAAACTGCCTG sequence TAAATAGACCTA isolate probe_HBV_ TTGATTGGAAAGTATGTCAGAGAATTG SEQIDNO:44 36Y18HCC,AB01439 012017_139 TGGGTCTTTTGGGCTTTGCTGCCCCTTT 5.1HepatitisBvirus TACACAATGTGGCTATCCTGCCTTAAT genomicDNA,complete GCCTTTATATGCATGTATACAATCTAA sequence GCAGGCTTTCA KR184660.1HepatitisB probe_HBV_ TCTCATCTGCCGGACCGTGTGCACTTC SEQIDNO:45 virusisolateSS_3_22, 012017_14 GCTTCACCTCTGCACGTCGCATGGAGA completegenome CCACCGTGAACGCCCATCAGGTCTTGC CCAAGGTCTTACATAAGAGGACTCTTG GACTCTCAGCAA isolate probe_HBV_ TGGCTATTGGCCATCAGCGCATGCGTG SEQIDNO:46 36Y18HCC,AB01439 012017_141 GAACCTTTGTGGCTCCTCTGCCGATCC 5.1HepatitisBvirus ATACTGCGGAACTCCTAGCAGCTTGTT genomicDNA,complete TTGCTCGCAGCCGGTCTGGAGCGAAAC sequence TGATCGGAACGG isolate probe_HBV_ ACAACTCTGTTGTTCTCTCTCGGAAAT SEQIDNO:47 36Y18HCC,AB01439 012017_142 ACACCTCCTTTCCATGGCTGCTAGGGT 5.1HepatitisBvirus GTGCTGCCAACTGGATCCTGCGCGGGA genomicDNA,complete CGTCCTTTGTTTACGTCCCGTCGGCGCT sequence GAATCCCGCGG isolate probe_HBV_ ACGACCCATCTCGGGGCCGTTTGGGTC SEQIDNO:48 36Y18HCC,AB01439 012017_143 TCTACCGTCCCCTTCTTCATCTGCCGTT 5.1HepatitisBvirus CCGGCCGACCACGGGGCGCACCTCTCT genomicDNA,complete TTACGCGGTCTCCCCGTCTGTGCCTTCT sequence CATCTGCCGG isolate probe_HBV_ ACCGTGTGCACTTCGCTTCACCTCTGC SEQIDNO:49 36Y18HCC,AB01439 012017_144 ACGTCGCATGGAGACCACCGTGAACG 5.1HepatitisBvirus CCCACCAGGTCTTGCCCAAGGTCTTAT genomicDNA,complete ATAAGAGGACTCTTGGACTCTCAGCAA sequence TGTCAACGACCGA isolate probe_HBV_ CCTTGAGGCATACTTCAAAGACTGTTT SEQIDNO:50 36Y18HCC,AB01439 012017_145 GTTTAAGGACTGGGAGGAGTTGGGGG 5.1HepatitisBvirus AGGAGTTTAGGTTAATGATCTTTGTAC genomicDNA,complete TAGGAGGCTGTAGGCATAAATTGGTCT sequence GTTCACCAGCACC isolate probe_HBV_ ATGCAACTTTTTCACCTCTGCCTAATCA SEQIDNO:51 36Y18HCC,AB01439 012017_146 TCTCATGTTCATGTCCTACTGTTCAAGC 5.1HepatitisBvirus CTCCAAGCTGTGCCTTGGGTGGCTTTG genomicDNA,complete GGGCATGGACATTGACCCGTATAAAG sequence AATTTGGAGCT isolate probe_HBV_ TCTGTGGAGTTACTCTCTTTTTTGCCTT SEQIDNO:52 36Y18HCC,AB01439 012017_147 CTGACTTCTTTCCTTCTATTCGAGATCT 5.1HepatitisBvirus CCTCGACACCGCCTCTGCTCTGTATCG genomicDNA,complete GGAGGCCTTAGAGTCTCCGGAACATTG sequence TTCACCTCAC isolate probe_HBV_ CATACAGCAATCAGGCAAGCTATTCTG SEQIDNO:53 36Y18HCC,AB01439 012017_148 TGTTGGGGTGAGTTGATGAATCTGGCC 5.1HepatitisBvirus ACCTGGGTGGGAAGTAATTTGGAAGA genomicDNA,complete CCCAGCATCCAGGGAATTAGTAGTCAG sequence CTATGTCAATGTT isolate probe_HBV_ AATATGGGCCTAAAAATCAGACAACT SEQIDNO:54 36Y18HCC,AB01439 012017_149 ACTGTGGTTTCACATTTCCTGTCTTACT 5.1HepatitisBvirus TTTGGAAGAGAAACTGTTCTTGAGTAT genomicDNA,complete TTGGTGTCTTTTGGAGTGTGGATTCGC sequence ACTCCTCCCGCT KR184660.1HepatitisB probe_HBV_ TGTCAACGTCCGACCTTGAGGCATACT SEQIDNO:55 virusisolateSS_3_22, 012017_15 TCAAAGACTGTTTGTTTAAGGACTGGG completegenome AGGAGTTGGGGGAGGAGATTAGGTTA AAGGTCTGGAGGCTGTAGGCATAAATT GGTCTGTTCACCA isolate probe_HBV_ TACAGACCACCAAATGCCCCTATCTTA SEQIDNO:56 36Y18HCC,AB01439 012017_150 TCAACACTTCCGGAAACTACTGTTGTT 5.1HepatitisBvirus AGACGACGAGGCAGGTCCCCTAGAAG genomicDNA,complete AAGAACTCCCTCGCCTCGCAGACGAAG sequence GTCTCAATCGCCG isolate probe_HBV_ CGTCGCAGAAGATCTCAATCTCGGGAA SEQIDNO:57 36Y18HCC,AB01439 012017_151 TCTCAATGTTAGTATCCCTTGGACTCAT 5.1HepatitisBvirus AAGGTGGGAAACTTTACTGGGCTTTAT genomicDNA,complete TCTTCTACTGTACCTGTCTTTAATCCTG sequence AGTGGCAAAC isolate probe_HBV_ TCCCTCCTTTCCTCACATTCATTTGCAG SEQIDNO:58 36Y18HCC,AB01439 012017_152 GAGGACATTATTAATAGATGTCAACAA 5.1HepatitisBvirus TATGTGGGCCCTCTTACAGTTAATGAA genomicDNA,complete AAAAGGAGATTAAAATTAATTATGCCT sequence GCTAGGTTCTA isolate probe_HBV_ TCCTAACCTTACCAAATATTTGCCCTTG SEQIDNO:59 36Y18HCC,AB01439 012017_153 GACAAAGGCATTAAACCATATTATCCT 5.1HepatitisBvirus GAACATGCAGTTCATCATTACTTCAAA genomicDNA,complete ACTAGGCATTATTTACATACTCTGTGG sequence AAGGCTGGCAT isolate probe_HBV_ TCTATATAAGAGAGAAACTACACGCA SEQIDNO:60 36Y18HCC,AB01439 012017_154 GCGCCTCATTTTGTGGGTCACCATATT 5.1HepatitisBvirus CTTGGGAACAAGAGCTACAGCAAACC genomicDNA,complete TCGACAAGGCATGGGGACAAATCTTTC sequence TGTTCCCAATCCTC isolate probe_HBV_ TGGGATTCTTTCCCGATCACCAGTTGG SEQIDNO:61 36Y18HCC,AB01439 012017_155 ACCCTGCGTTCGGAGCCAACTCAAACA 5.1HepatitisBvirus ATCCAGATTGGGACTTCAACCCCAACA genomicDNA,complete AGGATCACTGGCCAGAGGCAAATCAG sequence GTAGGAGCGGGAG isolate probe_HBV_ CTCCACCACATTCCACCAAGCTCTGCT SEQIDNO:62 22Y04HCC,AB01438 012017_156 ACACCCCAGAGTAAGGGGCCTATACTT 1.1HepatitisBvirus TCCTGCTGGTGGCTCCAGTTCCGGAAC genomicDNA,complete AGTAAACCCTGTTCCGACTACTGCCTC sequence TCCCATATCGTC isolate probe_HBV_ AATCTTCTCGAGGACTGGGGACCCTGC SEQIDNO:63 22Y04HCC,AB01438 012017_157 ACCGAACATGGAGAACACAACATCAG 1.1HepatitisBvirus GATTCCTAGGACCCCTGCTCGTGTTAC genomicDNA,complete AGGCGGGGTTTTTCTTGTTGACAAGAA sequence TCCTCACAATACC isolate probe_HBV_ ACAGAGTCTAGACTCGTGGTGGACTTC SEQIDNO:64 22Y04HCC,AB01438 012017_158 TCTCAATTTTCTAGGGGGAGCACCCAC 1.1HepatitisBvirus GTGTCCTGGCCAAAATTCGCAGTCCCC genomicDNA,complete AACCTCCAATCACTCACCAACCTCTTG sequence TCCTCCAATTTG isolate probe_HBV_ TCCTGGCTATCGCTGGATGTGTCTGCG SEQIDNO:65 22Y04HCC,AB01438 012017_159 GCGTTTTATCATATTCCTCTTCATCCTG 1.1HepatitisBvirus CTGCTATGCCTCATCTTCTTGTTGGTTC genomicDNA,complete TTCTGGACTACCAAGGTATGTTGCCCG sequence TTTGTCCTCT KR184660.1HepatitisB probe_HBV_ GCACCATGCAACTTTTTCACCTCTGCCT SEQIDNO:66 virusisolateSS_3_22, 012017_16 AATCATCTCATGTTCATGTCCTACTGTT completegenome CAAGCCTCCAAGCTGTGCCTTGGGTGG CTTTGGGGCATGGACATTGACCCGTAT AAAGAATTTG isolate probe_HBV_ ACTTCCAGGAACATCAACTACCAGCAC SEQIDNO:67 22Y04HCC,AB01438 012017_160 GGGACCATGCAAGACCTGCACGATTCC 1.1HepatitisBvirus TGCTCAAGGCACCTCTATGTTTCCCTCT genomicDNA,complete TGTTGCTGTACAAAACCTTCGGACGGA sequence AACTGCACTTG isolate probe_HBV_ TATTCCCATCCCATCATCCTGGGCTTTC SEQIDNO:68 22Y04HCC,AB01438 012017_161 GCAAGATTCCTATGGGAGTGGGCCTCA 1.1HepatitisBvirus GTCCGTTTCTCCTGGCTCAGTTTACTAG genomicDNA,complete TGCCATTTGTTCAGTGGTTCGTAGGGC sequence TTTCCCCCAC isolate probe_HBV_ TGTTTGGCTTTCAGTTATATGGATGAT SEQIDNO:69 22Y04HCC,AB01438 012017_162 GTGGTATTGGGGGCCAAGTCTGTACAA 1.1HepatitisBvirus CATCTTGAGTCCCTTTTTACCGCTGTTA genomicDNA,complete CCAATTTTCTTTTGTCTTTGGGTATACA sequence TTTGAACCCT isolate probe_HBV_ AATAAAACCAAACGTTGGGGTTACTCC SEQIDNO:70 22Y04HCC,AB01438 012017_163 CTTAACTTCATGGGATATGTAATTGGA 1.1HepatitisBvirus AGTTGGGGTACTTTACCGCAAGACCAT genomicDNA,complete ATTGTACTAAAAATCAAGCAATGTTTT sequence CGAAAACTGCCT isolate probe_HBV_ GTAAATAGACCTATTGATTGGAAAGTA SEQIDNO:71 22Y04HCC,AB01438 012017_164 TGTCAGAGAATTGTGGGTCTTTTGGGC 1.1HepatitisBvirus TTTGCTGCCCCTTTTACACAATGTGGCT genomicDNA,complete ATCCTGCCTTAATGCCTTTATATGCATG sequence TATACAATCT isolate probe_HBV_ AAGCAGGCTTTCACTTTCTCGCCAACT SEQIDNO:72 22Y04HCC,AB01438 012017_165 TACAAGGCCTTTCTGTGTAAACAATAT 1.1HepatitisBvirus CTGAACCTTTACCCCGTTGCCCGGCAA genomicDNA,complete CGGTCAGGTCTCTGCCAAGTGTTTGCT sequence GACGCAACCCCC isolate probe_HBV_ ACTGGATGGGGCTTGGCTATTGGCCAT SEQIDNO:73 22Y04HCC,AB01438 012017_166 CGCCGCATGCGTGGAACCTTTGTGGCT 1.1HepatitisBvirus CCTCTGCCGATCCATACTGCGGAACTC genomicDNA,complete CTAGCAGCTTGTTTTGCTCGCAGCCGG sequence TCTGGAGCGAAA isolate probe_HBV_ CTGATCGGAACGGACAACTCTGTTGTT SEQIDNO:74 22Y04HCC,AB01438 012017_167 CTCTCTCGGAAATACACCTCCTTTCCAT 1.1HepatitisBvirus GGCTGCTAGGGTGTGCTGCCAACTGGA genomicDNA,complete TCCTGCGCGGGACGTCCTTTGTTTACG sequence TCCCGTCGGCG isolate probe_HBV_ CTGAATCCCGCGGACGACCCATCTCGG SEQIDNO:75 22Y04HCC,AB01438 012017_168 GGCCGTTTGGGTCTCTACCGTCCCCTTC 1.1HepatitisBvirus TTCATCTGCCGTTCCGGCCGACCACGG genomicDNA,complete GGCGCACCTCTCTTTACGCGGTCTCCC sequence CGTCTGTGCCT isolate probe_HBV_ TCTCATCTGCCGGACCGTGTGCACTTC SEQIDNO:76 22Y04HCC,AB01438 012017_169 GCTTCACCTCTGCACGTCGCATGGAGA 1.1HepatitisBvirus CCACCGTGAACGCCCACCAGGTCTTGC genomicDNA,complete CCAAGGTCTTATATAAGAGGACTCTTG sequence GACTCTCAGCAA KR184660.1HepatitisB probe_HBV_ GAGCTTCTGTGGAGTTACTCTCTTTTTT SEQIDNO:77 virusisolateSS_3_22, 012017_17 GCCTTCTGACTTCTTTCCTTCCATTCGA completegenome GATCTCCTCGACACCGCCTCTGCTCTG TATCGGGAGGCCTTAGAGTCTCCGGAA CATTGTTCAC isolate probe_HBV_ TGTCAACGACCGACCTTGAGGCATACT SEQIDNO:78 22Y04HCC,AB01438 012017_170 TCAAAGACTGTTTGTTTAAGGACTGGG 1.1HepatitisBvirus AGGAGTTGGGGGAGGAGATTAGGTTA genomicDNA,complete ATGATCTTTGTACTAGGAGGCTGTAGG sequence CATAAATTGGTCT isolate probe_HBV_ GTTCACCAGCACCATGCAACTTTTTCA SEQIDNO:79 22Y04HCC,AB01438 012017_171 CCTCTGCCTAATCATCTCATGTTCATGT 1.1HepatitisBvirus CCTACTGTTCAAGCCTCCAAGCTGTGC genomicDNA,complete CTTGGGTGGCTTTAGGACATGGACATT sequence GACCCATATAA isolate probe_HBV_ AGAATTTGGAGCTTCTGTGGAGTTACT SEQIDNO:80 22Y04HCC,AB01438 012017_172 CTCTTTTTTGCCTTCTGACTTTTTTCCTT 1.1HepatitisBvirus CTATTCGAGATCTCCTCGACACCGCCT genomicDNA,complete CTGCTCTGTATCGGGAGGCCTTAGAGT sequence CTCCGGAACA isolate probe_HBV_ TTGTTCACCTCACCATACAGCACTCAG SEQIDNO:81 22Y04HCC,AB01438 012017_173 ACAAGCCATTCTGTGTTGGGGTGAGTT 1.1HepatitisBvirus GATGAATCTGGCCACCTGGGTGGGAA genomicDNA,complete GTAATTTGGAAGACCCAGCATCCAGGG sequence AATTAGTAGTCAG isolate probe_HBV_ CTATGTCAATGTTAATATGGGCCTAAA SEQIDNO:82 22Y04HCC,AB01438 012017_174 AATCAGACAACTACTGTGGTTTCACAT 1.1HepatitisBvirus TTCCTGTCTTACTTTTGGAAGAGAAAC genomicDNA,complete TGTTCTTGAGTATTTGGTGTCTTTTGGA sequence GTGTGGATTCG isolate probe_HBV_ CACTCCTCCTGCTTACAGACCATCAAA SEQIDNO:83 22Y04HCC,AB01438 012017_175 TGCCCCTATCTTATCAACACTTCCGGA 1.1HepatitisBvirus AACTACTGTTGTTAGACGACGAGGCAG genomicDNA,complete GTCCCCTAGAAGAAGAACTCCCTCGCC sequence TCGCAGACGAAG isolate probe_HBV_ GTCTCAATCGCCGCGTCGCAGAAGATC SEQIDNO:84 22Y04HCC,AB01438 012017_176 TCAATCTCGGGAACCTCAATGTTAGTA 1.1HepatitisBvirus TCCCTTGGACTCATAAGGTGGGAAACT genomicDNA,complete TTACTGGGCTTTATTCTTCTACTGTACC sequence TGTCTTTAATC isolate probe_HBV_ CTGAGTGGCAAACTCCCTCTTTTCCTC SEQIDNO:85 22Y04HCC,AB01438 012017_177 ATATTCATTTGCAGGAGGACATTATTA 1.1HepatitisBvirus ATAGATGTCAACAATATGTGGGCCCTC genomicDNA,complete TTACAGTTAATGAAAAAAGGAGATTA sequence AAATTAATTATGC isolate probe_HBV_ CTGCTAGGTTCTATCCTAACCTTACCA SEQIDNO:86 22Y04HCC,AB01438 012017_178 AATATTTGCCCTTGGACAAAGGCATTA 1.1HepatitisBvirus AACCATATTATCCGGAACATGCAGTTA genomicDNA,complete ATCATTACTTCAAAACTAGGCATTATT sequence TACATACTCTGT KR184660.1HepatitisB probe_HBV_ CTCACCATACAGCACTCAGGCAAGCTA SEQIDNO:87 virusisolateSS_3_22, 012017_18 TTCTCTGTTGGGGTGAGTTGATGAATC completegenome TGGCCACCTGGGTGGGAAGTAATTTGG AAGACCCAGCATCCAGGGATTTAGTAG TCAGCTATGTCA isolate probe_HBV_ ATGGGGACAAATCTTTCTGTTCCCAAT SEQIDNO:88 22Y04HCC,AB01438 012017_180 CCTCTGGGATTCTTTCCCGATCACCAG 1.1HepatitisBvirus TTGGACCCTGCGTTCGGAGCCAACTCA genomicDNA,complete AACAATCCAGATTGGGACTTCAACCCC sequence AACAAGGATCAC isolate probe_HBV_ TGGCCAGAGGCAAATCAGGTAGGAGC SEQIDNO:89 22Y04HCC,AB01438 012017_181 GGGAGCATTCGGGCCAGGGTTCACCCC 1.1HepatitisBvirus ACCACACGGCGGTCTTTTGGGGTGGAG genomicDNA,complete CCCTCAGGCTCAGGGCACATTGACAAC sequence AGTGCCAGTAGCA DQ683578.1HepatitisB probe_HBV_ CTCCACAACATTCCACCAAGCTCTGCT SEQIDNO:90 virusfromSouthKorea, 012017_182 AGATCCCAGAGTGAGGGGCCTATATTT completegenome TCCTGCTGGTGGCTCCAGTTCCGGAAC AGTAAACCCTGTTCCGACTACTGCCTC ACCCATATCGTC DQ683578.1HepatitisB probe_HBV_ AATCTTCTCGAGGACTGGGGACCCTGC SEQIDNO:91 virusfromSouthKorea, 012017_183 ACCGAACATGGAGAGCACAACATCAG completegenome GATTCCTAGGACCCCTGCTCGTGTTAC AGGCGGGGTTTTTCTTGTTGACAAGAA TCCTCACAATACC DQ683578.1HepatitisB probe_HBV_ ACAGAGTCTAGACTCGTGGTGGACTTC SEQIDNO:92 virusfromSouthKorea, 012017_184 TCTCAATTTTCTAGGGGGAGCACCCAC completegenome GTGTCCTGGCCAAAATTCGCAGTCCCC AACCTCCAATCACTCACCAACCTCTTG TCCTCCAATTTG DQ683578.1HepatitisB probe_HBV_ TCCTGGCTATCGCTGGATGTGTCTGCG SEQIDNO:93 virusfromSouthKorea, 012017_185 GCGTTTTATCATATTCCTCTTCATCCTG completegenome CTGCTATGCCTCATCTTCTTGTTGGTTC TTCTGGACTACCAAGGTATGTTGCCCG TTTGTCCTCT DQ683578.1HepatitisB probe_HBV_ ACTTCCAGGAACATCAACTACCAGCAC SEQIDNO:94 virusfromSouthKorea, 012017_186 GGGACCATGCAAGACCTGCACGATTCC completegenome TGCTCAAGGAACCTCTATGTTTCCCTCT TGTTGCTGTACAAAACCTTCGGACGGA AACTGCACTTG DQ683578.1HepatitisB probe_HBV_ TATTCCCATCCCATCATCCTGGGCTTTC SEQIDNO:95 virusfromSouthKorea, 012017_187 GTAAAATTCCTATGGGAGTGGGCCTCA completegenome GTCCGTTTCTCCTGGCTCAGTTTACTAG TGCCATTTGTTCAGTGGTTCGCAGGGC TTTCCCCCAC DQ683578.1HepatitisB probe_HBV_ TGTTTGGCTTTCAGTTATATGGATGAT SEQIDNO:96 virusfromSouthKorea, 012017_188 GTGGTATTGGGGGCCAAGTCTGTGCAA completegenome CATCTTGAGTCCCTTTTTACCTCTATTA CCAATTTTCTTTTGTCTTTGGGTATACA TTTGAACCCT DQ683578.1HepatitisB probe_HBV_ AATAAAACCAAACGTTGGGGCTACTCC SEQIDNO:97 virusfromSouthKorea, 012017_189 CTTAACTTCATGGGATATGTAATTGGA completegenome AGTTGGGGTACTTTACCACAGGAACAT ATTGTATTAAAACTCAAGCAATGTTTT CGGAAATTGCCT KR184660.1HepatitisB probe_HBV_ ATGTTAATATGGGCCTAAAAATCAGAC SEQIDNO:98 virusisolateSS_3_22, 012017_19 AACTATTGTGGTTTCACATTTCCTGTCT completegenome TACTTTTGGAAGAGAAACTGTTCTTGA GTATTTGGTGTCTTTTGGAGTGTGGATT CGCACTCCTC DQ683578.1HepatitisB probe_HBV_ GTAAATAGACCTATTGATTGGAAAGTA SEQIDNO:99 virusfromSouthKorea, 012017_190 TGTCAAAGAATTGTGGGTCTTTTGGAC completegenome TTTGCTGCCCCTTTTACACAATGTGGCT ATCCTGCATTGATGCCTTTATATGCAT GTATACAAGCT DQ683578.1HepatitisB probe_HBV_ AAGCAGGCTTTCACTTTCTCGCCAACT SEQIDNO:100 virusfromSouthKorea, 012017_191 TACAAGGCCTTTCTGTGTCAACAATAC completegenome CTGCACCTTTACCCCGTTGCCCGGCAA CGGTCAGGTCTCTGCCAAGTGTTTGCT GACGCAACCCCC DQ683578.1HepatitisB probe_HBV_ ACTGGATGGGGCTTGGCCATAGGCCAT SEQIDNO:101 virusfromSouthKorea, 012017_192 CGGCGCATGCGTGGAACCTTTGTGGCT completegenome CCTCTGCCGATCCATACTGCGGAACTC CTAGCGGCTTGTTTTGCTCGCAGCCGG TCTGGAGCAAAA DQ683578.1HepatitisB probe_HBV_ CTTATCGGGACCGACAACTCTGTTGTC SEQIDNO:102 virusfromSouthKorea, 012017_193 CTCTCTCGGAAATACACCTCCTTCCCA completegenome TGGCTGCTCGGGTGTGCTGCCAACTGG ATCCTGCGCGGGACGTCCTTTGTCTAC GTCCCGTCGGCG DQ683578.1HepatitisB probe_HBV_ CTGAATCCCGCGGACGACCCGTCTCGG SEQIDNO:103 virusfromSouthKorea, 012017_194 GGCCGTTTGGGCCTCTATCGTCCCCTTC completegenome TTCATCTGCCGTTCCAGCCGACCACGG GGCGCACCTCTCTTTACGCGGTCTCCC CGTCTGTGCCT DQ683578.1HepatitisB probe_HBV_ TCTCATCTGCCGGACCGTGTGCACTTC SEQIDNO:104 virusfromSouthKorea, 012017_195 GCTTCACCTCTGCACGTCGCATGGAAA completegenome CCACCGTGAACGCCCATCAGGTCTTGC CCAAGCTCTTACATAAGAGGACTCTTG GACTCTCAGCAA DQ683578.1HepatitisB probe_HBV_ TGTCAACGACCGACCTTGAGGCTTACT SEQIDNO:105 virusfromSouthKorea, 012017_196 TCAAAGACTGTTTGTTTAAAGACTGGG completegenome AGGAGTTGGGGGAGGAGACTAGGTTA AAGGTCTTTGTACTAGGAGGCTGTAGG CATAAATTGGTCT DQ683578.1HepatitisB probe_HBV_ GTTCACCAGCACCATGCAACTTTTTCA SEQIDNO:106 virusfromSouthKorea, 012017_197 CCTCTGCCTAATCATCTCATGTTCATGT completegenome CCTACTGTTCAAGCCTCCAAGCTGTGC CTTGGGTGGCTTTGGGGCATGGACATT GACCCGTATAA DQ683578.1HepatitisB probe_HBV_ AGAATTTGGAGCTTCTGCGGAGTTACT SEQIDNO:107 virusfromSouthKorea, 012017_198 CTCTTTTTTGCCTTCTGACTTCTTTCCTT completegenome CTATTCGAGATCTCCTCGACACCGCCT CTGCTCTATATCGGGAGGCCTTAGAGT CTCCGGAACA DQ683578.1HepatitisB probe_HBV_ TTGTTCACCTCACCATACAGCACTCAG SEQIDNO:108 virusfromSouthKorea, 012017_199 GCAAGCTATTCTGTGTTGGGGTGAGTT completegenome GATGAATCTGGCCACCTGGGTGGGAA GTAATTTGGAAGACCCAGCATCCAGGG AATTAGTAGTCAG KR184660.1HepatitisB probe_HBV_ AATCTTCTCGAGGACTGGGGACCCTGC SEQIDNO:109 virusisolateSS_3_22, 012017_2 ACCGAACATGGAGAGCACAACATCAG completegenome GATTCCTAGGACCCCTGCTCGTGTTAC AGGCGGGGTTTTTCTTGTTGACAAGAA TCCTCACAATACC KR184660.1HepatitisB probe_HBV_ CCGCTTACAGACCACCAAATGCCCCTA SEQIDNO:110 virusisolateSS_3_22, 012017_20 TCTTATCAACACTTCCGGAAACTACTG completegenome TTGTTAGACGACGAGGCAGGTCCCCTA GAAGAAGAACTCCCTCGCCTCGCAGAC GAAGGTCTCAAT DQ683578.1HepatitisB probe_HBV_ CTATGTCAATGTTAATATGGGCCTAAA SEQIDNO:111 virusfromSouthKorea, 012017_200 AATCAGACAACTATTGTGGTTTCACAT completegenome TTCCTGTCTTACTTTTGGAAGAGAAAC TGTTCTTGAGTATTTGGTGTCTTTTGGA GTGTGGATTCG DQ683578.1HepatitisB probe_HBV_ CACTCCTCCCGCTTACAGACCACCAAA SEQIDNO:112 virusfromSouthKorea, 012017_201 TGCCCCTATCTTATCAACACTTCCGGA completegenome AACTACTGTTGTTAGACGACGAGGCAG GTCCCCTAGAAGAAGAACTCCCTCGCC TCGCAGACGAAG DQ683578.1HepatitisB probe_HBV_ GTCTCAATCGCCGCGTCGCAGAAGATC SEQIDNO:113 virusfromSouthKorea, 012017_202 TCAATCTCGGGAATCTCAATGTTAGTA completegenome TCCCTTGGACTCATAAGGTGGGAAACT TTACTGGGCTTTATTCTTCTACTGTACC TGTCTCTAATC DQ683578.1HepatitisB probe_HBV_ CTGAGTGGCAAACTCCCTCCTTTCCTA SEQIDNO:114 virusfromSouthKorea, 012017_203 ACATTCATTTACAGGAGGACGTTATTA completegenome ATAGATGTCAACAATATGTGGGCCCTC TTACAGTTAATGAAAAAAGGAGATTA AAATTAATTATGC DQ683578.1HepatitisB probe_HBV_ CTGCTAGGTTCTATCCTAACCTTACCA SEQIDNO:115 virusfromSouthKorea, 012017_204 AATATTTGCCCTTGGATAAAGGCATTA completegenome AACCTTATTATCCTGAACATGCAGTTA ATCATTACTTCAAAACTAGGCATTATT TACATACTCTGT DQ683578.1HepatitisB probe_HBV_ GGAAGGCTGGCATTCTATATAAAAGA SEQIDNO:116 virusfromSouthKorea, 012017_205 GAAACTACACGCAGCGCTTCATTTTGT completegenome GGGTCACCATATTCTTGGGAACAAGAG CTACAGCATGGGAGGTTGGTCTTCCAA ACCTCGACAAGGC DQ683578.1HepatitisB probe_HBV_ ATGGGGACGAATCTTTCTGTTCCCAAT SEQIDNO:117 virusfromSouthKorea, 012017_206 CCTCTGGGATTCTTTCCCGATCACCAG completegenome TTGGACCCTGCGTTCAGAGCCAACTCA AACAATCCAGATTGGGACTTCAACCCC AACAAGGATCAC DQ683578.1HepatitisB probe_HBV_ TGGCCAGAGGCAAATCAGGTAGGAGC SEQIDNO:118 virusfromSouthKorea, 012017_207 GGGAGCATTCGGGCCAGGGTTCACCCC completegenome ACCACACGGCGGTCTTTTGGGGTGGAG CCCTCAGGCTCAGGGCATATTGACAAC TGTGCCAGCAGCG KR184660.1HepatitisB probe_HBV_ CATATTGACAACAGTGCCAGCAGCGCC SEQIDNO:119 virusisolateSS_3_22, 012017_208 TCCTCCTGCCTCCACCAATCGGCAGTC completegenome AGGAAGACAGCCTACTCCCATCTCTCC ACCTCTAAGAGACAGTCATCCTCAGGC CATGCAGTGGAA JN315779.1HepatitisB probe_HBV_ CATATTGACAACAGTGCCCGCAGCGCC SEQIDNO:120 virusgenotypeC2, 012017_209 TCCTCCTGCCTCCACCAATCGGCAGTT completegenome AGGAAGACAGCCTACTCCCATCTCTCC ACCTCTAAGAGACAGTCATCCTCAGGC CATGCAGTGGAA KR184660.1HepatitisB probe_HBV_ CGCCGCGTCGCAGAAGATCTCAATCTC SEQIDNO:121 virusisolateSS_3_22, 012017_21 GGGAATCTCAATGTTAGTATCCCTTGG completegenome ACTCATAAGGTGGGAAACTTTACTGGG CTTTATTCTTCTACTGTACCTGTCTTTA ATCCTGAGTGG GQ872211.1HepatitisB probe_HBV_ CATATTGACAACAGTGCCAGCAGCGCC SEQIDNO:122 virus,completegenome 012017_210 TCCTCCTGCCTCCACCAATCGGCAGTC AGGAAGACAGCCTACTCCCATCTCTCC ACCTCTAAGAGACAGTCATCCTCAGGC CATGCAGTGGAA D23680.1HepatitisB probe_HBV_ CATATTGACAACCGTGCCAGTAGCACC SEQIDNO:123 virus(B4-HBVST1) 012017_211 TCCTCCTGCCTCCACCAATCGGCAGTC completegenome AGGAAGACAGCCTACTCCCATCTCTCC sequence ACCTCTAAGAGACAGTCATCCTCAGGC CATGCAGTGGAA AY641559.1HepatitisB probe_HBV_ CATAGTGACACCAGTGCCAGCAGCGCC SEQIDNO:124 virusisolateHe53, 012017_212 TCCTCCTGCCTCCACCAATCGGCAGTC completegenome AGGAAGACAGCCTACTCCCATCTCTCC ACCTCTAAGAGACAGTCATCCTCAGGC CATGCAGTGGAA isolate probe_HBV_ GCATTCGGGCCAGGGTTCACCCCACCA SEQIDNO:125 36Y18HCC,AB01439 012017_213 CACGGCGGTCTTTTGGGGTGGAGCCCT 5.1HepatitisBvirus CAGGCTCAGGGTGCATTGACAACAGTG genomicDNA,complete CCAGTAGCACCTCCTCCTGCCTCCACC sequence AATCGGCAGCCT isolate probe_HBV_ CACATTGACAACAGTGCCAGTAGCACC SEQIDNO:126 22Y04HCC,AB01438 012017_214 TCCTCCTGCCTCCACCAATCGGCAGTC 1.1HepatitisBvirus AGGAAGACAGCCTACTCCCATCTCTCC genomicDNA,complete ACCTCTAAGAGACAGTCATCCTCAGGC sequence CATGCAGTGGAA DQ683578.1HepatitisB probe_HBV_ CATATTGACAACTGTGCCAGCAGCGCC SEQIDNO:127 virusfromSouthKorea, 012017_215 TCCTCCTGCCTCCACCAATCGGCAGTC completegenome AGAAAGACAGCCTACTCCCATCTCTCC ACCTCTAAGAGACAGTCATCCTCAGGC CATGCAGTGGAA KR184660.1HepatitisB probe_HBV_ CAAACTCCCTCCTTTCCTAACATTCATT SEQIDNO:128 virusisolateSS_3_22, 012017_22 TACAGGAAGACATTATTAATAGATGTC completegenome AACAATATGTGGGCCCTCTTACAGTTA ATGAAAAAAGGAGATTAAAATTAATT ATGCCTGCTAGG KR184660.1HepatitisB probe_HBV_ TTCTATCCTAACCTTACCAAATATTTGC SEQIDNO:129 virusisolateSS_3_22, 012017_23 CCTTGGATAAAGGCATTAAACCTTATT completegenome ATCCTGAACATGCAGTTAATCATTACT TCAAAACTAGGCATTATTTACATACTC TGTGGAAGGCT KR184660.1HepatitisB probe_HBV_ GGCATTCTATATAAAAGAGAAACTACA SEQIDNO:130 virusisolateSS_3_22, 012017_24 CGCAGCGCTTCATTTTGTGGGTCACCA completegenome TATTCTTGGGAACAAGAGCTACAGCAT GGGAGGTTGGTCTTCCAAACCTCGACA AGGCATGGGGAC KR184660.1HepatitisB probe_HBV_ GAATCTTTCTGTTCCCAATCCTCTGGG SEQIDNO:131 virusisolateSS_3_22, 012017_25 ATTCTTTCCCGATCACCAGTTGGACCC completegenome TGCGTTCGGAGCCAACTCAAACAATCC AGATTGGGACTTCAACCCCAACAAGG ATCACTGGCCAGA KR184660.1HepatitisB probe_HBV_ GGCAAATCAGGTAGGAGCGGGAGCAT SEQIDNO:132 virusisolateSS_3_22, 012017_26 TCGGGCCAGGGTTCACCCCACCACACG completegenome GCGGTCTTTTGGGGTGGAGCCCTCAGG CTCAGGGCATATTGACAACAGTGCCAG CAGCGCCTCCTCC JN315779.1HepatitisB probe_HBV_ CTCCACAACATTCCACCAAGCTCTGCT SEQIDNO:133 virusgenotypeC2, 012017_27 AGATCCCAGAGTGAGGGGCCTATATTT completegenome TCCTGCTGGTGGCTCCAGTTCCGGAAC AGTAAACCCTGTTCCGACTACTGCCTC ACCCATATCGTC JN315779.1HepatitisB probe_HBV_ AATCTTCTCGAGGACTGGGGACCCTGC SEQIDNO:134 virusgenotypeC2, 012017_28 ACCGAACATGGAGAACACAACATCAG completegenome GATTCCTAGGACCCCTGCTCGTGTTAC AGGCGGGGTTTTTCTTGTTGACAAGAA TCCTCACAATACC JN315779.1HepatitisB probe_HBV_ ACAGAGTCTAGACTCGTGGTGGACTTC SEQIDNO:135 virusgenotypeC2, 012017_29 TCTCAATTTTCTAGGGGAAGCACCCAC completegenome GTGTCCTGGCCAAAATTCGCAGTCCCC AACCTCCAATCACTCACCAACCTCTTG TCCTCCAATTTG KR184660.1HepatitisB probe_HBV_ ACAGAGTCTAGACTCGTGGTGGACTTC SEQIDNO:136 virusisolateSS_3_22, 012017_3 TCTCAATTTTCTAGGGGGAGCACCCAC completegenome GTGTCCTGGCCAAAATTCGCAGTCCCC AACCTCCAATCACTCACCAACCTCTTG TCCTCCAATTTG JN315779.1HepatitisB probe_HBV_ TCCTGGCTATCGCTGGATGTGTCTGCG SEQIDNO:137 virusgenotypeC2, 012017_30 GCGTTTTATCATATTCCTCTTCATCCTG completegenome CTGCTATGCCTCATCTTCTTGTTGGTTC TTCTGGACTACCAAGGTATGTTGCCCG TTTGTCCTCT JN315779.1HepatitisB probe_HBV_ ACTTCCAGGAACATCAACTACCAGCAC SEQIDNO:138 virusgenotypeC2, 012017_31 GGGACCATGCAAGACCTGCACGATTCC completegenome TGCTCAAGGAACCTCTATGTTTCCCTCT TGTTGCTGTACAAAACCTTCGGACGGA AACTGCACTTG JN315779.1HepatitisB probe_HBV_ TATTCCCATCCCATCATCCTGGGCTTTC SEQIDNO:139 virusgenotypeC2, 012017_32 GCAAAATTCCTATGGGAGTGGGCCTCA completegenome GTCCGTTTCTCCTGGCTCAGTTTACTAG TGCCATTTGTTCAGTGGTTCGCAGGGC TTTCCCCCAC JN315779.1HepatitisB probe_HBV_ TGTTTGGCTTTCAGTTATATGGATGAT SEQIDNO:140 virusgenotypeC2, 012017_33 GTGGTATTGGGGGCCAAGTCTGTACAA completegenome CATCTTGAGTCCCTTTTTACCTCTATTA CCAATTTTCTTTTGTCTTTGGGTATACA TTTGAACCCT JN315779.1HepatitisB probe_HBV_ AATAAAACCAAACGTTGGGGCTACTCC SEQIDNO:141 virusgenotypeC2, 012017_34 CTTAACTTCATGGGATATGTAATTGGA completegenome AGTTGGGGTACTTTACCACAGGAACAT ATTGTACTAAAAATCAAGCAATGTTTT CGGAAACTGCCT JN315779.1HepatitisB probe_HBV_ GTAAATAGACCTATTGATTGGAAAGTA SEQIDNO:142 virusgenotypeC2, 012017_35 TGTCAAAGAATTGTGGGTCTTTTGGGC completegenome TTTGCTGCCCCTTTTACACAATGTGGCT ATCCTGCCTTGATGCCTTTATATGCATG TATACAATCT JN315779.1HepatitisB probe_HBV_ AAGCAGGCTTTCACTTTCTCGCCAACT SEQIDNO:143 virusgenotypeC2, 012017_36 TACAAGGCCTTTCTGTGTAAACAATAT completegenome CTGCACCTTTACCCCGTTGCCCGGCAA CGGTCAGGTCTCTGCCAAGTGTTTGCT GACGCAACCCCC JN315779.1HepatitisB probe_HBV_ CTTATCGGGACTGACAACTCTGTTGTC SEQIDNO:144 virusgenotypeC2, 012017_38 CTCTCTCAGAAATACACCTCCTTCCCA completegenome TGGCTGCTCGGGTGTGCTGCCAACTGG ATCCTGCGCGGGACGTCCTTTGTCTAC GTCCCGTCGGCG KR184660.1HepatitisB probe_HBV_ TCCTGGCTATCGCTGGATGTGTCTGCG SEQIDNO:145 virusisolateSS_3_22, 012017_4 GCGTTTTATCATATTCCTCTTCATCCTG completegenome CTGCTATGCCTCATCTTCTTGTTGGTTC TTCTGGACTACCAAGGTATGTTGCCCG TTTGTCCTCT JN315779.1HepatitisB probe_HBV_ TCTCATCTGCCGGTCCGTGTGCACTTC SEQIDNO:146 virusgenotypeC2, 012017_40 GCTTCACCTCTGCACGTCGCATGGAGA completegenome CCACCGTGAACGCCCACCAGGTCTTGC CCAAGGTCTTACATAAGAGGACTCTTG GACTCTCAGCAA JN315779.1HepatitisB probe_HBV_ TGTCAACAACCGACCTTGAGGCATACT SEQIDNO:147 virusgenotypeC2, 012017_41 TCAAAGACTGTTTGTTTAAAGACTGGG completegenome AGGAGTTGGGGGAGGAGATTAGGTTA AAGGTCTTTGTACTAGGAGGCTGTAGG CATAAATTGGTCT JN315779.1HepatitisB probe_HBV_ GTTCACCAGCACCATGCAACTTTTTCA SEQIDNO:148 virusgenotypeC2, 012017_42 CCTCTGCCTAATCATCTCATGTTCATGT completegenome CCTACTGTTCAAGCCTCCAAGCTGTGC CTTGGGTGGCTTTGGGGCATGGACATT GACCCGTATAA JN315779.1HepatitisB probe_HBV_ AGAATTTGGAGCTTCTGTGGAGTTACT SEQIDNO:149 virusgenotypeC2, 012017_43 CTCTTTTTTGCCTTCTGACTTCTTTCCTT completegenome CTATTCGAGATCTCCTCGACACCGCCT CTGCTCTGTATCGGGAGGCCTTAGAGT CTCCGGAACA JN315779.1HepatitisB probe_HBV_ TTGTTCACCTCACCATACAGCACTCAG SEQIDNO:150 virusgenotypeC2, 012017_44 GCAAGCTATTCTGTGTTGGGGTGAGTT completegenome ATTGAATCTGGCCACCTGGGTGGGAAG TAATTTGGAAGACCCAGCATCCAGGGA ATTAGTAGTCAG JN315779.1HepatitisB probe_HBV_ CTATGTCAATGTTAATATGGGCCTAAA SEQIDNO:151 virusgenotypeC2, 012017_45 AATCAGACAACTATTGTGGTTTCACAT completegenome TTCCTGTCTTACTTTTGGAAGAGAAAC TGTTCTTGAGTATTTGGTGTCTTTTGGA GTGTGGATTCG JN315779.1HepatitisB probe_HBV_ GTCTCAATCGCCGCGTCGCCGAAGATC SEQIDNO:152 virusgenotypeC2, 012017_47 TCAATCTCGGGAATCTCAATGTTAGTA completegenome TCCCTTGGACTCATAAGGTGGGAAACT TTACTGGGCTTTATTCTTCTACTGTACC TGTCTTTAATC JN315779.1HepatitisB probe_HBV_ CTGAGTGGCAAACTCCCTCCTTTCCTA SEQIDNO:153 virusgenotypeC2, 012017_48 ACATTCATTTACAGGAGGACATTATTA completegenome ATAGATGTCAACAATATGTGGGCCCTC TCACAGTTAATGAAAAAAGGAGATTA AAATTAATTATGC KR184660.1HepatitisB probe_HBV_ ACTTCCAGGAACATCAACTACCAGCAC SEQIDNO:154 virusisolateSS_3_22, 012017_5 GGGACCATGCAAGACCTGCACGATTCC completegenome TGCTCAAGGAACCTCTATGTTTCCCTCT TGTTGCTGTACAAAACCTTCGGACGGA AACTGCACTTG JN315779.1HepatitisB probe_HBV_ ATGGGGACGAATCTTTCTGTTCCCAAT SEQIDNO:155 virusgenotypeC2, 012017_51 CCTCTGGGATTCTTTCCCGATCACCAG completegenome TTGGACCCTGCGTTCGGAGCCAACTCA AACAATCCAGATTGGGACTTCAACCCC AACAAGGATCAC JN315779.1HepatitisB probe_HBV_ TGGCCAGAGGCAAATCAGGTAGGAGC SEQIDNO:156 virusgenotypeC2, 012017_52 GGGAGCATTCGGGCCAGGGTTCACCCC completegenome ACCACACGGCGGTCTTTTGGGGTGGAG CCCTCAGGCTCAGGGCATATTGACAAC AGTGCCCGCAGCG GQ872211.1HepatitisB probe_HBV_ CTCCACAACATTCCACCAAGCTCTGCT SEQIDNO:157 virus,completegenome 012017_53 AGATCCCAGAGTGAGGGGCCTATATTT TCCTGCTGGTGGCTCCAGTTCCGGAAC AGTAAACCCTGTTCCGACTACTGCCTC ACCCATATCGTC GQ872211.1HepatitisB probe_HBV_ AATCTTCTCGAGGACTGGGGACCCTGC SEQIDNO:158 virus,completegenome 012017_54 ACCGAACATGGAGAGCACAACATCAG GATTCCTAGGACCCCTGCTCGTGTTAC AGGCGGGGTTTTTCTTGTTGACAAGAA TCCTCACAATACC GQ872211.1HepatitisB probe_HBV_ ACAGAGTCTAGACTCGTGGTGGACTTC SEQIDNO:159 virus,completegenome 012017_55 TCTCAATTTTCTAGGGGGAGCACCCAC GTGTCCTGGCCAAAATTCGCAGTCCCC AACCTCCAATCACTCACCAACCTCTTG TCCTCCAATTTG GQ872211.1HepatitisB probe_HBV_ TCCTGGCTATCGCTGGATGTGTCTGCG SEQIDNO:160 virus,completegenome 012017_56 GCGTTTTATCATATTCCTCTTCATCCTG CTGCTATGCCTCACCTTCTTGTTGGTCC TTCTGGACTACCAAGGTATGTTGCCCG TTTGTCCTCT GQ872211.1HepatitisB probe_HBV_ ACTTCCAGGAACATCAACTACCAGCAC SEQIDNO:161 virus,completegenome 012017_57 GGGACCATGCAAGACCTGCACGACTCC TGCTCAAGGAACCTCTATGTTTCCCTCT TGTTGCTGTACAAAACCTTCGGACGGA AACTGCACTTG GQ872211.1HepatitisB probe_HBV_ TATTCCCATCCCATCATCCTGGGCTTTC SEQIDNO:162 virus,completegenome 012017_58 GCAAGATTCCTATGGGAGTGGGCCTCA GTCCGTTTCTCCTGGCTCAGTTTACTAG TGCCATTTGTTCAGTGGTTCGCAGGGC TTTCCCCCAC GQ872211.1HepatitisB probe_HBV_ TGTTTGGCTTTCAGTTATATGGATGAT SEQIDNO:163 virus,completegenome 012017_59 GGGGTATTGGGGGCCAAGTCTGTACAA CATCTTGAGTCCCTTTTTACCTCTATTA CCAATTTTCTTTTGTCTTTGGGTATACA TTTGAACCCT KR184660.1HepatitisB probe_HBV_ TATTCCCATCCCATCATCCTGGGCTTTC SEQIDNO:164 virusisolateSS_3_22, 012017_6 GCAAGATTCCTATGGGAGTGGGCCTCA completegenome GTCCGTTTCTCCTGGCTCAGTTTACTAG TGCCATTTGTTCAGTGGTTCGTAGGGC TTTCCCCCAC GQ872211.1HepatitisB probe_HBV_ AATAAAACCAAACGTTGGGGCTACTCC SEQIDNO:165 virus,completegenome 012017_60 CTTAACTTCATGGGATATGTAATTGGA AGTTGGGGTACTTTACCACAGGAACAT ATTGTATTAAAAATCAAGAAATGTTTT CGGAAACTGCCT GQ872211.1HepatitisB probe_HBV_ GTAAATAGACCTATTGATTGGAAAGTA SEQIDNO:166 virus,completegenome 012017_61 TGTCAAAGAATTGTGGGTCTTTTGGGC TTTGCTGCCCCTTTTACACAATGTGGCT ATCCTGCCTTAATGCCTTTATATGCATG TATACAATCT GQ872211.1HepatitisB probe_HBV_ AAGCAGGCTTTCACTTTCTCGCCCACT SEQIDNO:167 virus,completegenome 012017_62 TACAAGGCCTTTCTGTGTCAACAATAC CTGCACCTTTACCCCGTTGCCCGGCAA CGGTCAGGTCTCTGCCAAGTGTTTGCT GACGCAACCCCC GQ872211.1HepatitisB probe_HBV_ ACTGGATGGGGCTTGGCCATAGGCCAT SEQIDNO:168 virus,completegenome 012017_63 CGGCGCATGCGTGGAACCTTTGTGGCT CCTCTGCCGATCCATACTGCGGAACTC CTAGCAGCTTGTTTTGCTCGCAGCCGG TCTGGAGCAAAA GQ872211.1HepatitisB probe_HBV_ CTTATCGGGACTGACAACTCTGTTGTC SEQIDNO:169 virus,completegenome 012017_64 CTCTCTCGGAAATACACCTCCTTCCCA TGGCTGCTCGGATGTGCTGCCAACTGG ATCCTGCGCGGGACGTCCTTTGTCTAC GTCCCGTCGGCG GQ872211.1HepatitisB probe_HBV_ CTGAATCCCGCGGACGACCCGTCTCGG SEQIDNO:170 virus,completegenome 012017_65 GGCCGTTTGGGCCTCTACCGTCCCCTT CTTCATCTGCCGTTCCAGCCGACCACG GGGCGCACCTCTCTTTACGCGGTCTCC CCGTCTGTGCCT GQ872211.1HepatitisB probe_HBV_ TCTCATCTGCCGGTCCGTGTGCACTTC SEQIDNO:171 virus,completegenome 012017_66 GCTTCACCTCTGCACGTCGCATGGAAA CCACCGTGAACGCCCACCAGGTCTTGC CCAAGGTCTTATATAAGAGGACTCTTG GACTCTCAGCAA GQ872211.1HepatitisB probe_HBV_ TGTCAACGACCGACCTTGAGGCATACT SEQIDNO:172 virus,completegenome 012017_67 TCAAAGACTGTTTGTTTAAAGACTGGG AGGAGTTGGGGGAGGAGATTAGGTTA ATGATCTTTGTACTAGGAGGCTGTAGG CATAAATTGGTCT GQ872211.1HepatitisB probe_HBV_ GTTCACCAGCACCATGCAACTTTTTCA SEQIDNO:173 virus,completegenome 012017_68 CCTCTGCCTAATCATCTCATGTTCATGT CCTACTGTTCAAGCCTCCAAGCTGTGC CTTGGGTGGCTTTGGGGCATGGACATT GACCCGTATAA GQ872211.1HepatitisB probe_HBV_ AGAATTTGGAGCTTCTGCGGAGTTACT SEQIDNO:174 virus,completegenome 012017_69 CTCTTTTTTGCCTTCTGACTTCTTTCCG TCTATTCGAGATCTCCTCGACACCGCC TCTGCTCTGTATAGGGAGGCCTTAGAG TCTCCGGAACA KR184660.1HepatitisB probe_HBV_ TGTTTGGCTTTCAGTTATATGGATGAT SEQIDNO:175 virusisolateSS_3_22, 012017_7 GTGGTATTGGGGGCCAAGTCTGTACAA completegenome CATCTTGAGTCCCTTTTTACCTCTATTA CCAATTTTCTTGTGTCTTTGGGTATACA TTTGAACCCT GQ872211.1HepatitisB probe_HBV_ TTGTTCACCTCACCATACAGCACTCAG SEQIDNO:176 virus,completegenome 012017_70 GCAAGCTATTCTGTGTTGGGGTGAGTT GATGAATCTGGCCACCTGGGTGGGAA GTAATTTGGAAGACCCAGCATCCAGGG AATTAGTAGTCGG GQ872211.1HepatitisB probe_HBV_ CTATGTCAATGTTAATATGGGCCTAAA SEQIDNO:177 virus,completegenome 012017_71 ACTCAGACAACTATTGTGGTTTCACAT TTCCTGTCTTACTTTTGGAAGAGAAAC TGTTCTTGAGTATTTGGTGTCTTTTGGA GTGTGGATTCG GQ872211.1HepatitisB probe_HBV_ CACTCCTACCGCTTACAGACCACCAAA SEQIDNO:178 virus,completegenome 012017_72 TGCCCCTATCTTATCAACACTTCCGGA AACTACTGTTGTTAGACGACGAGGCAG GTCCCCTAGAAGAAGAACTCCCTCGCC TCGCAGACGAAG GQ872211.1HepatitisB probe_HBV_ GTCTCAATCGCCGCGTCGCAGAAGATC SEQIDNO:179 virus,completegenome 012017_73 TCAATCTCGGGAATCTCAATGTTAGTA TCCCTTGGACTCATAAGGTGGGAAACT TTACTGGGCTTTATTCTTCTACTGTACC TGTCTTTAATC GQ872211.1HepatitisB probe_HBV_ CTGAGTGGCAAACTCCCTCCTTTCCTA SEQIDNO:180 virus,completegenome 012017_74 ACATTCATTTACAGGAGGACATTATTA ATAGATGTCAACAATATGTGGGCCCTC TTACAGTTAATGAAAAAAGGAGATTA AAATTAATTATGC GQ872211.1HepatitisB probe_HBV_ CTGCTAGGTTCTATCCTAACCTTACCA SEQIDNO:181 virus,completegenome 012017_75 AATATTTGCCCTTGGATAAGGGCATTA AACCTTATTATCCTGAACATGCAGTTA ATCATTACTTCAAAACTAGGCATTATT TACATACTCTGT GQ872211.1HepatitisB probe_HBV_ GGAAGGCTGGCATTCTATATAAAAGA SEQIDNO:182 virus,completegenome 012017_76 GAAACTACACGCAGCGCTTCATTTTGT GGGTCACCATATTCTTGGGAACAAGAG CTACAGCATGGGAGGTTGGTCTTCCAA ACCTCGAAAAGGC GQ872211.1HepatitisB probe_HBV_ ATGGGGACGAATCTTTCTGTTCCCAAT SEQIDNO:183 virus,completegenome 012017_77 CCTCTGGGATTCTTTCCCGATCACCAG TTGGACCCTGCATTCGGAGCCAACTCA AACAATCCAGATTGGGACTTCAACCCC AACAAGGATCAC GQ872211.1HepatitisB probe_HBV_ TGGCCAGAGGCAACTCAGGTAGGAGC SEQIDNO:184 virus,completegenome 012017_78 GGGAGCATTCGGGCCAGGGTTCACCCC ACCACACGGCGGTCTTTTGGGGTGGAG CCCTCAGGCTCAGGGCATATTGACAAC AGTGCCAGCAGCG D23680.1HepatitisB probe_HBV_ CTCCACAACATTCCACCAAGCTCTGCT SEQIDNO:185 virus(B4-HBVST1) 012017_79 AGACCCCAGAGTGAGGGGCCTATACTT completegenome TCCTGCTGGTGGCTCCAGTTCCGGAAC sequence AGTAAACCCTGTTCCGACTACTGCCTC ACCCATATCGTC KR184660.1HepatitisB probe_HBV_ AATAAAACCAAACGTTGGGGCTACTCC SEQIDNO:186 virusisolateSS_3_22, 012017_8 CTTAACTTCATGGGATATGTAATTGGA completegenome AGTTGGGGTACTTTACCACAGGAACAT ATTGTACAAAAACTCAAGCAATGTTTT CGGAAACTGCCT D23680.1HepatitisB probe_HBV_ AATCTTCTCGAGGACTGGGGACCCTGC SEQIDNO:187 virus(B4-HBVST1) 012017_80 ACCGAACATGGAGAACACAACATCAG completegenome GATTCCTAGGACCCCTGCTCGTGTTAC sequence AGGCGGGGTTTTTCTTGTTGACAAGAA TCCTCACAATACC D23680.1HepatitisB probe_HBV_ ACAGAGTCTAGACTCGTGGTGGACTTC SEQIDNO:188 virus(B4-HBVST1) 012017_81 TCTCAATTTTCTAGGGGGAGCACCCAC completegenome GTGTCCTGGCCAAAATTCGCAGTCCCC sequence AACCTCCAATCACTCACCAACCTCTTG TCCTCCAATTTG D23680.1HepatitisB probe_HBV_ ACCTGGCTATCGCTGGATGTGTCTGCG SEQIDNO:189 virus(B4-HBVST1) 012017_82 GCGTTTTATCATATTCCTCTTCATCCTG completegenome CTGCTATGCCTCATCTTCTTGTTGGTTC sequence TTCTGGACTACCAAGGTATGTTGCCCG TTTGTCCTCT D23680.1HepatitisB probe_HBV_ ACTTCCAGGAACATCAACTACCAGCAC SEQIDNO:190 virus(B4-HBVST1) 012017_83 AGGACCATGCAAGACCTGCACGATTCC completegenome TGCTCAAGGAACCTCTATGTTTCCCTCT sequence TGTTGCTGTACAAAACCTTCGGACGGA AACTGCACTTG D23680.1HepatitisB probe_HBV_ TATTCCCATCCCATCATCCTGGGCTTTC SEQIDNO:191 virus(B4-HBVST1) 012017_84 GCAAGATTCCTATGGGAGTGGGCCTCA completegenome GTCCGTTTCTCCTGGCTCAGTTTACTAG sequence TGCCATTTGTTCAGTGGTTCGTAGGGC TTTCCCCCAC D23680.1HepatitisB probe_HBV_ TGTTTGGCTTTCAGTTATATGGATGAT SEQIDNO:192 virus(B4-HBVST1) 012017_85 GTGGTATTGGGGGCCAAGTCTGTACAA completegenome CATCTTGAGTCCCTTTTTACCTCTATTA sequence CCCATTTTCTTTTATCTTTGGGTATACA TTTGAACCCC D23680.1HepatitisB probe_HBV_ AATAAAACCAAACGTTGGGGCTACTCC SEQIDNO:193 virus(B4-HBVST1) 012017_86 CTTAACTTCATGGGATATGTAATTGGA completegenome TGTTGGGGTACTTTACCGCAAGAACAT sequence ATTGTACTAAAAATCAAGCAATGTTTT CGAAAACTGCCT D23680.1HepatitisB probe_HBV_ GTAAATAGACCTATTGATTGGAAAGTA SEQIDNO:194 virus(B4-HBVST1) 012017_87 TGTCAGAGAATTGTGGGTCTTTTGGGC completegenome TTTGCTGCCCCTTTTACACAATGTGGCT sequence ATCCTGCCTTAAAGCCTTTATATGCAT GTATACAAGCT D23680.1HepatitisB probe_HBV_ AAGCAGGCTTTCACTTTCTCGCCGACT SEQIDNO:195 virus(B4-HBVST1) 012017_88 TACAAGGCCTTTCTGTGTAAACAATAT completegenome CTGAACCTTTACCCCGTTGCCCGGCAA sequence CGGTCAGGTCTCTGCCAAGTGTTTGCT GACGCAACCCCC D23680.1HepatitisB probe_HBV_ ACTGGCTGGGGCTTGGCTATCGGCCAT SEQIDNO:196 virus(B4-HBVST1) 012017_89 CGCCGCATGCGTGGAACCTTTGTGGCT completegenome CCTCTGCCGATCCATACTGCGGAACTC sequence CTAGCAGCTTGTTTTGCTCGCAGCCGG TCTGGAGCGAAA KR184660.1HepatitisB probeHBV_ GTAAATAGACCTATTGACTGGAAAGTA SEQIDNO:197 virusisolateSS_3_22, 012017_9 TGTCAAAGAATTGTGGGTCTTTTGGGC completegenome TTTGCTGCCCCTTTTACACAATGTGGCT ATCCTGCCTTGATGCCTTTATATGCATG TATACAAGCT D23680.1HepatitisB probe_HBV_ CTTATCGGCACCGACAACTCTGTTGTC SEQIDNO:198 virus(B4-HBVST1) 012017_90 CTCTCTCGGAAATACACCTCATTTCCA completegenome TGGCTGCTAGGGTGTGCTGCCAACTGG sequence ATCCTGCGCGGGACGTCCTTTGTCTAC GTCCCGTCGGCG D23680.1HepatitisB probe_HBV_ CTGAATCCCGCGGACGACCCGTCTCGG SEQIDNO:199 virus(B4-HBVST1) 012017_91 GGCCGTTTGGGACTCTACCGTCCCCTT completegenome CTTCATCTGCCGTTCCGGCCAACCACG sequence GGGCGCACCTCTCTTTACGCGGTCTCC CCGTCTGTGCCT D23680.1HepatitisB probe_HBV_ TCTCATCTGCCGGGCCGTGTGCACTTC SEQIDNO:200 virus(B4-HBVST1) 012017_92 GCTTCACCTCTGCACGTCGCATGGAAA completegenome CCTCCGTGAACGCCCACCAGGTCTTGC sequence CCAAGGTCTTATATAAGAGGACTCTTG GACTCTCAGCGA D23680.1HepatitisB probe_HBV_ TGTCAACGACCGACCTTGAGGCATACT SEQIDNO:201 virus(B4-HBVST1) 012017_93 TCAAAGACTGTTTGTTTAAGGACTGGG completegenome AGGAGTTGGGGGAGGTACTAGGAGGC sequence TGTAGGCATAAATTGGTCTGTTCACCA GCACCATGCAACT D23680.1HepatitisB probe_HBV_ TTTTCACCTCTGCCTAATCATCTCATGT SEQIDNO:202 virus(B4-HBVST1) 012017_94 TCATGTCCTACTGTTCAAGCCTCCAAG completegenome CTGTGCCTTGGGTGGCTTTGGGGCATG sequence GACATTGACCCGTATAAAGAATTTGGA GCTTCTGTGGA D23680.1HepatitisB probe_HBV_ GTTACTCTCTTTTTTGCCTTCTGACTTC SEQIDNO:203 virus(B4-HBVST1) 012017_95 TTTCCTTCTATTCGAGATCTCCTCGACA completegenome CCGCCTCAGCTCTGTATCGGGAGGCCT sequence TAGAGTCTCCGGAACATTGTTCTCCTC ACCATACAGC D23680.1HepatitisB probe_HBV_ ACTCAGGCAAGCTATTCTGTGTTGGGG SEQIDNO:204 virus(B4-HBVST1) 012017_96 TGAGTTGATGAATCTGGCCACCTGGGT completegenome GGGAAGTAATTTGGAAGACCCAGCAT sequence CCAGGGAATTAGTAGTCAGCTATGTCA ATGTTAATATGGG D23680.1HepatitisB probe_HBV_ CCTAAAAATCAGACAACTACTGTGGTT SEQIDNO:205 virus(B4-HBVST1) 012017_97 TCACATTTCCTGTCTTACTTTTGGAAGA completegenome GAAACTGTTCTTGAGTATTTGGTGTCTT sequence TTGGAGTGTGGATTCGCACTCCTCCTG CTTACAGACC D23680.1HepatitisB probe_HBV_ ACCAAATGCCCCTATCTTATCAACACT SEQIDNO:206 virus(B4-HBVST1) 012017_98 TCCGGAAACTACTGTTGTTAGACGACG completegenome AGGCAGGTCCCCTAGAAGAAGAACTC sequence CCTCGCCTCGCAGACGAAGGTCTCAAT CGCCGCGTCGCAG D23680.1HepatitisB probe_HBV_ AAGATCTCAATCTCGGGAATCTCAATG SEQIDNO:207 virus(B4-HBVST1) 012017_99 TTAGTATCCCTTGGACTCATAAGGTGG completegenome GAAACTTTACTGGGCTTTATTCTTCTAC sequence TGTACCTGTCTTTAATCCTGAGTGGCA AACTCCCTCCT isolate probe_HBV_ CACCAAGCTCTGATAGACCCCAGAGTA SEQIDNO:208 36Y18HCC,AB01439 012017_a_1 AGGGGCCTATACTTTCCTGCTGGTGGC 5.1HepatitisBvirus TCCAGTTCCGGAACAGTAAACCCTGTT genomicDNA,complete CCGACTACTGCCTCACCCATATCGTCA sequence ATCTTCTCGAGG isolate probe_HBV_ CTTTCTCGCCAACTTACAAGGCCTTTCT SEQIDNO:209 36Y18HCC,AB01439 012017_a_2 GTGTAAACAATATCTGAACCTTTACCC 5.1HepatitisBvirus CGTTGCTCGGCAACGGTCAGGTTTATG genomicDNA,complete CCAAGTGTTTGCTGACGCAACCCCCAC sequence TGGATGGGGCT isolate probe_HBV_ GGAAGGCAGGCATTCTATATAAGAGA SEQIDNO:210 22Y04HCC,AB01438 012017_a_3 GAAACTACACGCAGCGCCTCATTTTGT 1.1HepatitisBvirus GGGTCACCATATTCTTGGGAACAAGAG genomicDNA,complete CTACAGCATGGGAGGTTGGTCTTCCAA sequence ACCTCGACAAGGC JN315779.1HepatitisB probe_HBV_ ACTGGATGGGGCTTGGCCATAGGCCAT SEQIDNO:211 virusgenotypeC2, 012017_a_4 CAGCGCATGCGTGGAACCTTTGTGGCT completegenome CCTCTGCCGATCCATACTGCGGAACTC ATAGAAGCTTGTTTTGCTCGCAGCCGG TCTGGAGCGAAA JN315779.1HepatitisB probe_HBV_ CTGAATCCCGCGGACGACCCGTCTCGG SEQIDNO:212 virusgenotypeC2, 012017_a_5 GACCGTTTGGGCCTCTACCGTCCCCTT completegenome CTTCATCTGCCGTTCCGGCCGACCACG GGGCGCACCTCTCTTTACGCGGTCTCC CCGTCTGTGCCT JN315779.1HepatitisB probe_HBV_ CACTCCTACCGCTTACAGACCACCAAA SEQIDNO:213 virusgenotypeC2, 012017_a_6 TGCCCCTATCTTATCAACACTTCCGGA completegenome AACTACTGTTGTTAGACGACGAGGCAG GTCCCCTAGAAGAAGAACTCCCTCGCC TCGCAGACGAAG JN315779.1HepatitisB probe_HBV_ CTGCTAGGTTCTATCCTAACCATACCA SEQIDNO:214 virusgenotypeC2, 012017_a_7 AATATTTGCCCTTGGATAAAGGCATTA completegenome AACCTTATTATCCTGAACATGTAGTTA ATCATTACTTCAAAACTAGGCATTATT TACATACTTTGG JN315779.1HepatitisB probe_HBV_ GGAAGGCTGGCATTCGGTATAAGAGA SEQIDNO:215 virusgenotypeC2, 012017_a_8 GAAACTACACGCAGCGCTTCATTTTGT completegenome GGGTCACCATATTCTTGGGAACAAGAG CTACAGCATGGGAGGTTGGTCTTCCAA ACCTCGACAAGGC

    Example 2: Next-Generation Sequencing Analysis for Detection of HBV Insertion Site

    [0052] 2-1. DNA Shearing

    [0053] 1) Extract genomic DNA from liver tissue of a patient with hepatitis and crush (sonication) it into nucleotides of about 100 to 120 base pairs in length. After diluting 1 g of gDNA passed through Quality Control (QC) on a 96-well plate with 60 L, transfer it to a Covaris strip tube and seal with sealing tape.

    [0054] 2) Transfer the strip tube to a steel rack and mount it on a device.

    [0055] 3) As Table 3 below, shear it after setting Covaris (Covaris LE200).

    TABLE-US-00003 TABLE 3 Duty Factor 30 PIP, W 400 Cycles per Burst 200 Time (seconds) 100 Temperature 5 to 9 C.

    [0056] 2) Sample Purification

    [0057] 1) Transfer the sheared sample into a new 1.5 mL tube.

    [0058] 2) Place 90 L of AMPure beads, vortex it for 5 seconds, and perform incubation at room temperature for 5 minutes.

    [0059] 3) Place the sample in a magnetic particle concentrator (MPC), and after 3 minutes, discard the supernatant.

    [0060] 4) Add 200 L of 70% ethanol while the sample is in MPC, and after 1 minute, discard the supernatant (repeat twice).

    [0061] 5) Completely dry the beads (5 minutes to 10 minutes).

    [0062] 6) Remove the sample tube from MPC, add 50 L of nuclease-free water, and resuspend AMPure beads.

    [0063] 7) After incubating at room temperature for 2 minutes to 3 minutes, spin it down.

    [0064] 8) Place the sample in MPC, and after 2 minutes, transfer 48 L of the supernatant into a new 1.5 mL tube.

    [0065] 2-3. Repairing the Ends

    [0066] 1) After mixing all of the components of Table 4 below, lid off in PCR and perform at 20 C. for 30 minutes.

    TABLE-US-00004 TABLE 4 Component Volume DNA sample 48 L Water 35.2 L End repair buffer 10 L dNTP mix 1.6 L T4 DNA polymerase 1 L Klenow DNA polymerase 2 L T4 PNK 2.2 L Total 100 L

    [0067] 2) Sample Purification

    [0068] {circle around (1)} Place the sample performed in 3. 1) above into a new 1.5 mL tube.

    [0069] {circle around (2)} Place 180 L of AMPure beads (1.8), vortex it for 5 seconds, and perform incubation for 5 minutes at room temperature.

    [0070] {circle around (3)} Place the sample in a magnetic particle concentrator (MPC), and after 3 minutes, discard the supernatant.

    [0071] {circle around (4)} While the sample is in MPC, add 200 L of 70% ethanol, and after 1 minute, discard the supernatant (repeat twice).

    [0072] {circle around (5)} Completely dry the beads (5 minutes to 10 minutes).

    [0073] {circle around (6)} Remove the sample tube from MPC, add 32 L of nuclease-free water, and resuspend AMPure beads.

    [0074] {circle around (7)} After incubating for 2 minutes to 3 minutes at room temperature, spin it down.

    [0075] {circle around (8)} Place the sample in MPC, and after 2 minutes, transfer 30 L of the supernatant into a new 1.5 mL tube.

    [0076] 2-4. Addition of A Base to the 3 End of DNA Fragment

    [0077] 1) After adding all of the components of Table 5 below, lid off in PCR and perform at 37 C. for 30 minutes.

    TABLE-US-00005 TABLE 5 Component Volume DNA sample 30 L Water 11 L 10X Klenow DNA polymerase buffer 5 L dATP 1 L Klenow exo(3 to 5 exo minus) 3 L Total 50 L

    [0078] 2) Sample Purification

    [0079] {circle around (1)} Place the sample performed in 4. 1) above into a new 1.5 mL tube.

    [0080] {circle around (2)} Place 180 L of AMPure beads (1.8), vortex it for 5 seconds, and perform incubation for 5 minutes at room temperature.

    [0081] {circle around (3)} Place the sample in a magnetic particle concentrator (MPC), and after 3 minutes, discard the supernatant.

    [0082] {circle around (4)} While the sample is in MPC, add 200 L of 70% ethanol, and after 1 minute, discard the supernatant (repeat twice).

    [0083] {circle around (5)} Completely dry the beads (5 minutes to 10 minutes).

    [0084] {circle around (6)} Remove the sample tube from MPC, add 15 L of nuclease-free water, and resuspend AMPure beads.

    [0085] {circle around (7)} After incubating for 2 minutes to 3 minutes at room temperature, spin it down.

    [0086] {circle around (8)} Place the sample in MPC, and after 2 minutes, transfer 13 L of the supernatant into a new 1.5 mL tube.

    [0087] 2-5. Adapter Ligation to DNA Fragment

    [0088] 1) After adding all of the components of Table 6 below, lid off in PCR and perform at 20 C. for 15 minutes.

    TABLE-US-00006 TABLE 6 Component Volume DNA sample 13 L Water 15.5 L 5X T4 DNA ligase buffer 10 L Adapter oligo mix 10 L T4 DNA ligase 1.5 L Total 50 L

    [0089] 2) Sample Purification

    [0090] {circle around (1)} Place the sample performed in 2-5. 1) above into a new 1.5 mL tube.

    [0091] {circle around (2)} Place 180 L of AMPure beads (1.8), vortex it for 5 seconds, and perform incubation for 5 minutes at room temperature.

    [0092] {circle around (3)} Place the sample in a magnetic particle concentrator (MPC), and after 3 minutes, discard the supernatant.

    [0093] {circle around (4)} While the sample is in MPC, add 200 L of 70% ethanol, and after 1 minute, discard the supernatant (repeat twice).

    [0094] {circle around (5)} Completely dry the beads (5 minutes to 10 minutes).

    [0095] {circle around (6)} Remove the sample tube from MPC, add 17 L of nuclease-free water, and resuspend AMPure beads.

    [0096] {circle around (7)} After incubating for 2 minutes to 3 minutes at room temperature, spin it down.

    [0097] {circle around (8)} Place the sample in MPC, and after 2 minutes, transfer 15 L of the supernatant into a new 1.5 mL tube.

    [0098] 2-6. Amplification of Adapter-Ligated Library

    [0099] 1) Prepare components in Table 7 below.

    TABLE-US-00007 TABLE 7 Component Volume Index Adapter-ligated library 15 L Water 21 L SureSelect primer 1.0 (Forward) 1.25 L SureSelect Indexing Pre-Capture PCR(Reverse) 1.25 L Primer Herculase 5X Reaction Buffer 10 L dNTP mix 0.5 L Herculase II polymerase 1 L Total 50 L

    [0100] 2) Amplify according to the Pre-LM PCR program below.

    TABLE-US-00008 TABLE 8 Step PCR step Time Step 1. 98 C. 2 mins Step 2. 98 C. 30 s Step 3. 65 C. 30 s Step 4. 72 C. 1 min Step 5. Repeat Steps 2 to 4 for 6 times Step 6. 72 C. 10 minutes Step 7. 4 C. Hold

    [0101] 2-7. Sample Purification

    [0102] {circle around (1)} Transfer the sample passed through the steps above into a new 1.5 mL tube.

    [0103] {circle around (2)} Place 180 L of AMPure beads (1.8), vortex it for 5 seconds, and perform incubation for 5 minutes at room temperature.

    [0104] {circle around (3)} Place the sample in a magnetic particle concentrator (MPC), and after 3 minutes, discard the supernatant.

    [0105] {circle around (4)} While the sample is in MPC, add 200 L of 70% ethanol, and after 1 minute, discard the supernatant (repeat twice).

    [0106] {circle around (5)} Completely dry the beads (5 minutes to 10 minutes).

    [0107] {circle around (6)} Remove the sample tube from MPC, add 17 L of nuclease-free water, and resuspend AMPure beads.

    [0108] {circle around (7)} After incubating for 2 minutes to 3 minutes at room temperature, spin it down.

    [0109] {circle around (8)} Place the sample in MPC, and after 2 minutes, transfer 15 L of the supernatant into a new 1.5 mL tube.

    [0110] 2-8. Assessment of Quality and Quantity

    [0111] In order to confirm whether the library size was made within the intended range to optimize the efficiency of hybridization and to confirm the concentration to check if the amount at which hybridization could be attempted was achieved, the size and concentration of a library were measured using Agilent 4200 Tape Station and D1000 Screen Tape, and the result was shown in FIG. 2. As shown in FIG. 2, peaks having a DNA library size of about 250 bp to 350 bp were mostly observed.

    [0112] 2-9. Hybridization

    [0113] 1) Drill a hole in a 1.5 mL tube lid and dispense 200 ng or more and 500 ng or less of the prepped library.

    [0114] 2) Completely dry using SpeedVac (45 C.) (60 minutes).

    [0115] 3) After making a block mix as below, place 5.6 L each into a dried tube, vortex lightly, and resuspend the library (prepped library).

    TABLE-US-00009 TABLE 9 Component Volume Pre-LM sample 500 ng 3.4 L SureSelect Block #1 (green cap) 2.5 L SureSelect Block #2 (blue cap) 2.5 L SureSelect Block #3 (brown cap) 0.6 L Total 9 L

    [0116] 4) After making a hybridization buffer with the composition of Table 10 below, dispense 0.2 mL into a PCR tube.

    TABLE-US-00010 TABLE 10 Component Volume SureSelect Hyb #1 6.63 L SureSelect Hyb #2(red) 0.27 L SureSelect Hyb #3(yellow) 2.65 L SureSelect Hyb #4 3.45 L Total 13 L

    [0117] 5) Perform RNase block dilution as Table 11 below.

    TABLE-US-00011 TABLE 11 Capture Library Size RNase Block dilution 3.0 Mb or more 25% (1:3) 3.0 Mb or less 10% (1:9)

    [0118] 6-1) The volume used for hybridization is different depending on the total size of a probe. It is because the concentration of the probe itself is different. Since the volume is different, the dilution ratio and the used volume of the RNase block should be different. The final concentration of RNase block is the same as 6-1 and 6-2. As a result, in the case of a general bait of 3 MB or more, it is applied to a large-sized probe targeting the entire exome.

    TABLE-US-00012 TABLE 12 Component Volume Hybridization Buffer mixture from step 4 13 L 25% RNase Block solution from step 5-1, 5-2 2 L Capture Library 3 Mb 5 L Total 20 L

    [0119] 6-2) The volume used for hybridization is different depending on the total size of a probe. It is because the concentration of the probe itself is different. Since the volume is different, the dilution ratio and the used volume of the RNase block should be different. The final concentration of RNase block is the same as 6-1 and 6-2. In the case of a general bait of 3 MB or less, it was applied in this experiment.

    TABLE-US-00013 TABLE 13 Component Volume Hybridization Buffer mixture from step 4 13 L 10% RNase Block solution from step 5-1, 5-2 5 L Capture Library 3 Mb 2 L Total 20 L

    [0120] 7) For gDNA library+block mix plate or a strip tube (prepped library), set up the PCR program as below and perform.

    TABLE-US-00014 TABLE 14 PCR program Time Lid temperature: 105 C. 95 C. 5 minutes 65 C. Hold

    [0121] 8) When the temperature of a prepped library (an entire set of libraries made available for NGS sequencing of gDNA samples used in the experiment) sample reaches 65 C., place the prepped library sample in a capture library (a set of probes including a target area of the size of 120 nt) and a hybridization mix (a reagent (buffer) to enable hybridization conditions) prepared above, and mix well by pipetting up and down for 3 to 5 times.

    [0122] 9) Close the lid well and hybridize for 24 hours at 65 C. (lid 105 C.) (up to 72 hours is possible).

    [0123] 2-10. Preparation of Magnetic Beads

    [0124] 1) Preheat SureSelect Wash Buffer #2 in a water bath (65 C.).

    [0125] 2) Vortex well Dynal MyOne Streptavidin T1 (Invitrogen) magnetic beads.

    [0126] 3) Dispense 50 L per sample into a 1.5 mL tube.

    [0127] 4) Wash the beads as the following.

    [0128] a. Place 200 L of SureSelect Binding buffer and vortex lightly.

    [0129] b. After spinning down, place it in DynaMag-2 device for 1 minute, and remove the supernatant.

    [0130] c. Repeat the above process for a total of 3 times.

    [0131] 5) Resuspend the beads washed in 200 L of SureSelect Binding buffer.

    [0132] 2-11. Hybridization Capture Selection with SureSelect

    [0133] 1) After mixing a hybridization mixture and a bead solution, mount it on a rotator and perform a reaction at room temperature for 30 minutes (check if the sample in the tube is mixed well).

    [0134] 2) After spinning down, place it in DynaMag-2 device for 3 minutes and remove the supernatant. 3) Place 200 L of SureSelect Wash Buffer #1 and vortex until the beads are completely resuspended.

    [0135] 4) Incubate at room temperature for 15 minutes. Lightly vortex every 5 minutes to mix the beads well.

    [0136] 5) After spinning down, place it in DynaMag-2 device for 3 minutes and remove the supernatant.

    [0137] 6) Wash the beads as the following.

    [0138] a. Place 200 L of prewarmed SureSelect Wash Buffer #2 and vortex until the beads are completely resuspended.

    [0139] b. Incubate at 65 C. for 10 minutes. Lightly vortex every 5 minutes to mix the beads well.

    [0140] c. After spinning down, place it in DynaMag-2 device and remove the supernatant.

    [0141] d. Repeat the above process for a total of 3 times. 7) Place 30 L of nuclease-free water in MPC and resuspend.

    [0142] 2-12. Addition of Index Tags by Amplification After Hybridization (Post-Hybridization)

    [0143] 1) Prepare reagents as in Table 15 below.

    TABLE-US-00015 TABLE 15 Reagent Volume Captured DNA 30 L Water 6.5 L Herculase 5X Reaction Buffer 10 L dNTP mix(25 mM each) 0.5 L Herculase II DNA polymersase 1 L SureSelect Indexing Post-Capture PCR (Forward) Primer 1 L Index PCR (reverse) primer 1 L Total 50 L

    [0144] 2) Perform amplification according to the PCR program below.

    TABLE-US-00016 TABLE 16 Step PCR step Time Step 1. 98 C. 1 min Step 2. 98 C. 20 s Step 3. 57 C. 1 min Step 4. 72 C. 1 min Step 5. Repeat steps 2 to 4 for 11 times Step 6. 72 C. 10 minutes Step 7. 4 C. Hold

    [0145] 2-13. Purification of Sample Using Agencourt AMPure XP beads

    [0146] 1) Vortex 50 L of the amplified DNA library and 90 L of AMPure beads (1.8) and mix.

    [0147] 2) Incubate at room temperature for 5 minutes.

    [0148] 3) Place the sample in a magnetic particle concentrator (MPC), and after 3 minutes, discard the supernatant.

    [0149] 4) Add 500 L of 70% ethanol while the sample is in MPC, and after 1 minute, discard the supernatant (repeat twice).

    [0150] 5) Completely dry the beads (5 minutes to 10 minutes).

    [0151] 6) Remove the sample tube from MPC, add 15 L of nuclease-free water, and resuspend AMPure beads.

    [0152] 7) After incubating at room temperature for 2 minutes to 3 minutes, spin it down.

    [0153] 8) Place the sample in MPC, and after 2 minutes, place 30 L of the supernatant into a new 1.5 mL tube.

    [0154] 2-14. Confirmation of Library

    [0155] It is a library state after hybridization has been performed and only a target region has been amplified. In FIG. 2, only the target region was selected from the entire library, and the library size was increased by 50 bp at once while adding an index and the like during the amplification process. It is the library at the final stage for sequencing, and in order to finally confirm whether sequencing is possible (determining whether or not a library is made normally), the size and state of the DNA library are confirmed using Agilent 4200 TapeStation and D1000 ScreenTape, and the concentration of the DNA library is confirmed using qPCR. As shown in FIG. 3, the library size has peaks of 250 bp to 350 bp.

    [0156] 2-15. Analysis of HBV Gene Insertion Site

    [0157] The sequenced reads were mapped to the reference sequence (HBV+Human genome) to create a BAM file, which is a binary of the Sequence Alignment map (SAM) file. Among the mapped reads, the chimeric read that was split-mapped to HBV and the human genome was selected to identify break points. Next, for each point, a region that satisfied read count >10, average mapping quality (MQ) >20 was defined as an HBV-human integration site, and the location of HBV and the human genome was searched. Recurrently inserted human genes were collected, gene-annotation was performed and analyzed to discover the overall biological function of each gene, and the results were shown in FIG. 4. As shown in FIG. 4, it was found that the HBV virus was inserted into the overall human whole genome, and in particular, it was confirmed that the insertion rate was high in the TERT protomer region of chromosome number 5. Through this, it is possible to comprehensively infer the effect of HBV insertion on the human genome. HBV insertion is an important direct tumor-inducing phenomenon in the occurrence of liver cancer, and understanding of an insight into its biological action is required, but there is little understanding of HBV insertion until now. Meanwhile, the NGS technique has become available to identify non-biased insertion sites therefor, but whole genome sequencing (WGS, full-length genome sequencing) is very difficult to use in clinical practice due to its cost limitations. The present invention described above is a sequencing method that applies the existing NGS technique targeting HBV inserted in the human genome, and since it is a high-depth sequencing analysis that is more efficient than WGS and can detect more HBV insertion sites at a cost of about of the WGS method, it is considered that the academic-clinical value thereof will be very high in the future compared to its cost-effectiveness.