Metagenomics-based respiratory tract throat swab sample library building method and pathogen detection method

Abstract

The invention provides a metagenomics-based respiratory tract throat swab sample library building method and a pathogen detection method, and relates to the technical field of microbiological detection. The library building method comprises the following steps: (a) pretreatment of a to-be-detected sample solution: adding lysozyme for digestion; (b) extraction of total nucleic acid of the sample: not adding carrier RNA in an extraction process; (c) cDNA synthesis: carrying out reverse transcription by adopting a random primer; (d) purifying a product synthesized in step (c) by using magnetic beads; and (e) construction of a sample DNA library. Based on a DNA-RNA co-sequencing principle, the invention provides the metagenomics-based method suitable for throat swab sample library building and detection and pathogen detection of a throat swab sample metagenome can be realized through a single reaction.

Description

technical field [0001] The invention relates to the technical field, in particular to a metagenomics-based method for building a library of respiratory and throat swab samples and a method for detecting pathogens. Background technique [0002] There are various types of pathogens that cause respiratory infections, including viruses, bacteria, fungi, etc. Most of them use DNA as the carrier of genetic information, but some viruses (such as influenza virus, coronavirus, etc.) use RNA as the carrier of genetic information. Metagenome sequencing of respiratory clinical samples has now become an important method for the detection of respiratory infection pathogens. By directly detecting all microbial nucleic acids in the sample, it can not only detect multiple pathogens in the sample at the same time, but also discover some conventional methods. New or rare pathogens that are difficult to identify. Due to some differences in the sequencing methods for DNA or RNA, the current con...

AI Technical Summary

Problems solved by technology

However, in many cases in actual work, it may not be possible to judge based on experience that the pathogens causing the infection are bacteria, viruses or other types of pathogens, and once the wrong library construction method is selected, the pathogens may be missed.

On the other hand, throat swab is a conventional non-invasive respiratory sample, which is often used in the detection of respiratory pathogens, but the nucleic acid content in the sample is generally low. Time and cost, and the nucleic acid content is generally difficult to meet the second library construction

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