Molecular-based diagnostic kits and methods for detecting nucleotide sequences of DNA, DNA copy and RNA of infectious agents, transgenes, alleles or non-encoding sequences, with no initial isolation are provided. The kit comprises at least a first calibrated dropper bottle with an anti-contamination system containing a solution of polymerase enzyme, chaotropic agents, salts and deoxyribonucleotides; a second calibrated dropper bottle with an anti-contamination system containing a set of at least 4 primers, part of the sequences being preserved in the nucleic acid fragment to be detected; a dropper bottle containing a mixture of developing reagents; a carrier to hold the sample; at least one LFD developing system, colorimetric or fluorogenic reaction; positive and negative controls. The polymerase enzyme is selected from Bst enzymes formed by DNA polymerase/helicase, with no exonuclease activity. The set of primers is designed from gene sequences to be detected, whether of deoxyribonucleic or ribonucleic nature.

Molecular-based diagnostic kits and methods for detecting nucleotide sequences of DNA, DNA copy and RNA of infectious agents, transgenes, alleles or non-encoding sequences, with no initial isolation are provided. The kit comprises at least a first calibrated dropper bottle with an anti-contamination system containing a solution of polymerase enzyme, chaotropic agents, salts and deoxyribonucleotides; a second calibrated dropper bottle with an anti-contamination system containing a set of at least 4 primers, part of the sequences being preserved in the nucleic acid fragment to be detected; a dropper bottle containing a mixture of developing reagents; a carrier to hold the sample; at least one LFD developing system, colorimetric or fluorogenic reaction; positive and negative controls. The polymerase enzyme is selected from Bst enzymes formed by DNA polymerase/helicase, with no exonuclease activity. The set of primers is designed from gene sequences to be detected, whether of deoxyribonucleic or ribonucleic nature.

The disclosure provides methods for characterizing a target DNA present in a sample. The methods involve contacting the sample with one or more universal primers to amplify target DNA; contacting the amplified target DNA with a type V CRISPR/Cas effector protein and one or more guide RNAs, where the contacting generates a cleavage product comprising a 5′ overhang; and ligating a double-stranded nucleic acid adapter to the cleavage product, to generate a ligation product. The ligation product includes the target DNA, which can be sequenced. The sample can be subjected to one or more amplification steps prior to the contacting step, with primers that provide for amplification of nucleic acids of, e.g., specific pathogens, categories of pathogens, two or more different pathogens, or two or more different categories of pathogens.

The disclosure provides methods for characterizing a target DNA present in a sample. The methods involve contacting the sample with one or more universal primers to amplify target DNA; contacting the amplified target DNA with a type V CRISPR/Cas effector protein and one or more guide RNAs, where the contacting generates a cleavage product comprising a 5′ overhang; and ligating a double-stranded nucleic acid adapter to the cleavage product, to generate a ligation product. The ligation product includes the target DNA, which can be sequenced. The sample can be subjected to one or more amplification steps prior to the contacting step, with primers that provide for amplification of nucleic acids of, e.g., specific pathogens, categories of pathogens, two or more different pathogens, or two or more different categories of pathogens.

A method and kit related thereto are described for the collection and maintenance of detectability of a plurality of species of microbiological agents in a single clinical sample as well as an integral method for handling a plurality of the samples and managing information associated therewith for reporting a sum of diagnostic results for each sample.

The invention is directed to compositions, kits, and methods for amplifying and detecting a Zika virus nucleic acid sequence in a sample, which comprises a variety of combinations of forward oligonucleotide primers, reverse oligonucleotide primers, and oligonucleotide probes.

The present invention relates to methods for improving multiplex real-time PCR assays by the use of an internal control probe labeled with Quasar 705.

A method of reducing host nucleic acids in a biological sample and a use thereof, which belong to the technical field of gene detection is provided. The method includes the steps of a) performing pre-treatment for the biological sample under a mild condition with partial lysis and/or no lysis of host cell membrane, thereby obtaining a liquid sample; and b) taking the liquid sample followed by adding a nucleic acid digestion reagent, thereby degrading the host nucleic acid which is exposed in the liquid sample. The method is used for reducing host nucleic acids in a biological sample and non-selectively enriching the targets, such as bacteria, fungi, viruses, mycoplasma/chlamydia.

Methods for the rapid detection of the presence or absence of Trichomonas vaginalis (TV) in a biological or non-biological sample are described. The methods can include performing an amplifying step, a hybridizing step, and a detecting step. Furthermore, primers, probes targeting the target TV gene, along with kits are provided that are designed for the detection of TV.

There is described herein a method for specifically detecting Babesia species nucleic acid in a sample, which in one aspect comprises: (1) contacting a sample, said sample suspected of containing Babesia species nucleic acid, with at least two oligomers for amplifying a target region of a Babesia species target nucleic acid, wherein the at least two amplification oligomers comprise: (a) a first amplification oligomer comprising a first target-hybridizing sequence (i) that is from about 15 to about 33 contiguous nucleotides in length, is contained in the sequence of SEQ ID NO:66 and comprises SEQ ID NO:56 or 57; or (ii) that is from about 15 to about 33 contiguous nucleotides in length, is contained in the sequence of SEQ ID NO:96 and comprises SEQ ID NO:101; or (iii) that is from about 15 to about 33 contiguous nucleotides in length, is contained in the sequence of SEQ ID NO:97 and comprises SEQ ID NO:101; (iv) comprises or consists of SEQ ID NO:8; (v) comprises or consists of SEQ ID NO:83 and (b) a second amplification oligomer comprising a second target-hybridizing sequence that is from about 15 to about 33 contiguous nucleotides in length, and (i) is contained in SEQ ID NO:68 and comprises SEQ ID NO:52, SEQ ID NO:53, SEQ ID NO:54, SEQ ID NO:55, or SEQ ID NO:85; or (ii) is contained in SEQ ID NO:67 and comprises SEQ ID NO:45 or SEQ ID NO:52; or (iii) is contained in SEQ ID NO:70 and comprises SEQ ID NO:46, SEQ ID NO:47, SEQ ID NO:48, SEQ ID NO:49, SEQ ID NO:50, or SEQ ID NO:51; (2) performing an in vitro nucleic acid amplification reaction, wherein any Babesia target nucleic acid present in said sample is used as a template for generating an amplification product; and (3) detecting the presence or absence of the amplification product, thereby indicating the presence or absence of Babesia species target nucleic acid in said sample.