Detection and identification of microbial species uses liquid chromatography as a bulk filtration process for rapid isolation and collection of microbial RNA from a biological specimen. In various embodiments, gene sequencing of microbial RNA molecules from a biological specimen is enhanced by obtaining and then preparing the biological specimen as a test sample for liquid chromatography that is used to bulk filter microbial RNA molecules from a mixture of RNA molecules in the test sample to isolate and collect the microbial RNA molecules in two or more fraction outputs, wherein at least one of the two or more fraction outputs is a fraction within a void volume of the liquid chromatography, preparing one or more fraction outputs for gene sequencing, including the fraction output that is within the void volume, and conducting gene sequencing on the one or more prepared outputs to detect microbial RNA from the biological specimen.

A manually actuated chromatography device comprising a chamber for receiving a liquid sample, a pump with a metering valve, and a chromatography element, wherein the pump moves a predetermined volume of liquid from the sample chamber to the chromatography element.

A manually actuated chromatography device comprising a chamber for receiving a liquid sample, a pump with a metering valve, and a chromatography element, wherein the pump moves a predetermined volume of liquid from the sample chamber to the chromatography element.

A fractional concentration of fetal relevant DNA in a mixture of DNA from a biological sample is determined based on amounts of DNA fragments of a particular size or range of sizes. DNA fragments may be sequenced to obtain sequence reads, and the sequence reads may be aligned to a reference genome to determine sizes of the DNA fragments. Calibration data points (e.g., as a calibration function) indicate a correspondence between values of a parameter providing a statistical measure of a size profile and the fractional concentration of the fetal DNA. For a given sample, a value of the parameter can be determined from DNA fragments of a particular size or range of sizes in a sample. A comparison of the value to the calibration data points can provide the estimate of the fractional concentration of the fetal DNA.

A fractional concentration of fetal relevant DNA in a mixture of DNA from a biological sample is determined based on amounts of DNA fragments of a particular size or range of sizes. DNA fragments may be sequenced to obtain sequence reads, and the sequence reads may be aligned to a reference genome to determine sizes of the DNA fragments. Calibration data points (e.g., as a calibration function) indicate a correspondence between values of a parameter providing a statistical measure of a size profile and the fractional concentration of the fetal DNA. For a given sample, a value of the parameter can be determined from DNA fragments of a particular size or range of sizes in a sample. A comparison of the value to the calibration data points can provide the estimate of the fractional concentration of the fetal DNA.

The present invention relates to a nucleic acid detection kit and a nucleic acid detection method, which use nanoparticles. More specifically, the present invention relates to: a nucleic acid detection method comprising a step of amplifying and labeling nucleic acids, and then capturing the same by using nanoparticles and centrifuging the same; and a nucleic acid detection kit using the method. The present invention is effective since a negative or positive determination for a particular disease can be made, through the nucleic acid detection method not comprising a separation step, in a more rapid, simple, sensitive, and highly reliable manner.

The present invention relates to a nucleic acid detection kit and a nucleic acid detection method, which use nanoparticles. More specifically, the present invention relates to: a nucleic acid detection method comprising a step of amplifying and labeling nucleic acids, and then capturing the same by using nanoparticles and centrifuging the same; and a nucleic acid detection kit using the method. The present invention is effective since a negative or positive determination for a particular disease can be made, through the nucleic acid detection method not comprising a separation step, in a more rapid, simple, sensitive, and highly reliable manner.

Diverse cDNA libraries derived from cell-free mRNA and methods of preparing the same are provided. The library may be prepared by extracting RNA from a bodily fluid such as serum or plasma, separating the RNA from contaminants, synthesizing cDNA with a reverse transcriptase enzyme, and enriching protein-coding nucleotide sequences. The library may include a multiplicity of transcripts from solid tissues. Cf-RNA can be measured by qPCR, sequencing, or other suitable methods.

Diverse cDNA libraries derived from cell-free mRNA and methods of preparing the same are provided. The library may be prepared by extracting RNA from a bodily fluid such as serum or plasma, separating the RNA from contaminants, synthesizing cDNA with a reverse transcriptase enzyme, and enriching protein-coding nucleotide sequences. The library may include a multiplicity of transcripts from solid tissues. Cf-RNA can be measured by qPCR, sequencing, or other suitable methods.

A method is provided to extract genetic material of a virus, for example the SARS-CoV-2 virus, from a wastewater sample. The wastewater sample is in the range of 0.5 to 5 mL in volume. The wastewater sample may be prefiltered, but is not separated to concentrate the virus in a solids fraction. A lysis buffer is added to the sample, followed by a waiting period of at least one minute, followed by the addition of an alcohol. 1.5-4.5 mg of magnetic beads are added per mL of wastewater sample. The magnetic beads are later separated from the sample. Genetic material is eluted from the magnetic beads, optionally at a temperature of at least 45° C. A test kit is provided for use in performing the method.