Primer group and kit for detecting 27 respiratory tract pathogens based on nucleic acid mass spectrometry and application of primer group and kit

Abstract

The invention provides a primer group capable of simultaneously detecting 27 respiratory tract pathogens based on a nucleic acid mass spectrometry technology, a kit and application, and belongs to thefield of pathogen nucleic acid detection. According to the kit, PCR amplification primers and single-base extension primers are designed for 27 clinically common respiratory tract pathogens. Pathogenic bacteria corresponding primer and single base extension primer sequences are shown in SEQ ID NO: 1-81. According to the nucleic acid mass spectrometry, a multiple RT-PCR technology and a single base extension technology are integrated, pathogen target genes are amplified through multiple RT-PCR, and the detection sensitivity is improved; in the single-base extension reaction, one base amplification is carried out on an extension primer, and pathogen identification is carried out by distinguishing the molecular mass of an extension product, so that the specificity is high. Meanwhile, the flux of the nucleic acid mass spectrum is high, amplification of 40 genes can be achieved at most through one reaction hole, and 384 samples can be analyzed at most at a time by means of the chip technology; the kit has the advantages of high automation degree, high detection flux, high sensitivity and good specificity.

Description

technical field [0001] The invention belongs to the field of pathogenic microorganism detection, and in particular relates to a primer set, a kit and an application thereof for detecting 27 kinds of respiratory pathogens based on nucleic acid mass spectrometry. Background technique [0002] Respiratory tract infection is a relatively common infectious disease clinically, especially in infants, the elderly, and immunocompromised patients, often leading to severe symptoms and even death. According to the WHO report, about 3 million people died of lower respiratory tract infection in 2016. Common respiratory infection pathogens include viruses, bacteria, mycoplasma, chlamydia and other microorganisms. Viral pathogens mainly include influenza A virus, influenza B virus, parainfluenza virus, respiratory syncytial virus, rhinovirus, metapneumovirus, adenovirus, etc. Bacterial pathogens include Streptococcus pneumoniae, Haemophilus influenzae, and Moraxella catarrhalis. Most resp...

AI Technical Summary

Problems solved by technology

Multiplex PCR detection has relatively high sensitivity and specificity, but is limited by the number of fluorescent types in each reaction. Generally, one reaction well can only detect 3-4 pathogens. If more pathogens need to be detected, more Reaction wells, so that the convenience of detection and the detection throughput are greatly reduced

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