High-throughput sequencing detection method for pathogenic microorganisms with full-process quality control

Abstract

The invention provides a high-throughput sequencing detection method for pathogenic microorganisms with full-process quality control. The method comprises the following steps: adding an internal reference into a first to-be-detected microorganism sample to obtain a second to-be-detected microorganism sample; extracting DNA in the second to-be-detected microorganism sample, a preset positive control sample and a blank control sample respectively; constructing a sequencing library for the extracted DNA samples by adopting a pathogenic microorganism detection kit respectively; sequencing the sequencing library based on a second-generation sequencing platform, performing quality control processing on original sequencing data in a sequencing result, and removing a sequence compared to a host genome to obtain to-be-compared data; and comparing the to-be-compared data based on a pre-established comparison database, and determining main pathogenic microorganisms according to a comparison result. Rapid clinical detection of the pathogenic microorganisms which do not need to be predicted, do not have preference and have high throughput for infectious diseases is realized, meanwhile, full-process quality control of a system is realized, and false positive is reduced.

Description

technical field [0001] The invention relates to the technical field of clinical detection of infectious diseases, in particular to a high-throughput sequencing detection method for pathogenic microorganisms in the whole process of quality control. Background technique [0002] Traditional diagnostic techniques for pathogenic microorganisms in infectious diseases include microscopy, culture, and serological methods. Microscopy is less sensitive unless the sample has a high disease burden and is not specific. The sensitivity of culture varies with pathogens, especially some difficult-to-culture microorganisms with complex nutritional requirements, such as cryptococcus, the most common 3 neuroinvasive fungi can be cultured on standard, variable sensitivity bacterial media , as for more rare fungi, specific media are required; for example, herpes simplex virus can only be cultured from CSF in up to 5% of cases of herpes simplex virus encephalitis, and antibiotic treatment can a...

AI Technical Summary

Problems solved by technology

However, there is currently no systematic, full-process quality control mNGS solution for the detection of pathogenic microorganisms for clinical needs

Images