Multiple PCR detection kit for pathogens of respiratory tract, application and application method thereof

Abstract

The invention provides a multiple PCR detection kit for pathogens of respiratory tract, an application and an application method thereof. The PCR detection kit comprises a mixture of various primers and probes, a PCR reaction liquid, a PCR enzyme liquid, an inner quality control product and a nucleic acid extraction reagent. The detection kit is respiratory syncytial virus used for detecting one or more pathogens of influenza a virus, influenza B virus, respiratory syncytial virus, parainfluenza I, parainfluenza II, parainfluenza III, adenovirus, human metapneumovirus and enterovirus or rhinovirus. The application of the kit comprises the following steps: firstly, carrying out sampling; then carrying out PCR amplification and detection; and finally, obtaining a detection result. By combingand coordinating the nucleic acid extraction reagent, the primer pairs, the probes, a PCR program, the PCR reaction liquid and the PCR enzyme liquid, the technical effects of shortening the detectiontime and being good in detection sensitivity and good in specificity are achieved finally.

Description

technical field [0001] The invention belongs to the technical field of nucleic acid amplification, and mainly relates to a kit for detecting respiratory pathogens, its application and its application method. Background technique [0002] Respiratory tract infection is very common in clinical practice, but due to the variety of pathogens that cause respiratory tract infection, rapid and accurate identification of pathogens is of great significance for clinical diagnosis and treatment and prevention of drug abuse. Fluorescent quantitative PCR technology based on Taqman probes can specifically amplify a large number of pathogenic nucleic acid molecules, and at the same time use the accumulation of fluorescent signals to realize the interpretation of negative and positive results. Compared with biochemical indicators and immune methods, molecular diagnosis is extremely sensitive, and can identify pathogen types and subtypes based on the differences in gene sequences of different...

AI Technical Summary

Problems solved by technology

[0003] However, several problems with traditional molecular diagnostic methods have hindered the large-scale use of this method in respiratory viral infections

Firstly, the detection process is relatively long, and the molecular diagnosis operation is complicated, involving nucleic acid extraction and product amplification. How to simplify the operation process and save experimental time is of great significance for patients to receive correct treatment quickly; secondly, most of the current molecular diagnostic tests are single-plex However, there are many types of respiratory viruses. There are about 145 serotypes of the most common rhinovirus that causes the common cold alone. The single-plex PCR system often has negative test results for patients, so it cannot give patients effective treatment. At present, there are many methods for detecting respiratory pathogens at home and abroad, each of which has its own advantages and disadvantages. The detection technologies include traditional isolation and culture methods, immunological detection technologies, and emerging molecular biology technologies. Nucleic acid-based molecular detection The method has become a revolutionary technology for the detection of respiratory pathogens due to its rapidity, sensitivity, specificity, and time-saving and labor-saving characteristics. Multiplex PCR refers to adding multiple pairs of specific primers in the same reaction system. At the same time, how to make the added primers Not interfering with each other and its detection sensitivity is not lower than that of a single PCR detection has become a major technical difficulty, and in critical or emergency situations, how to improve the efficiency of the entire detection process so that patients can receive timely and accurate treatment has become an emergency Therefore, it has clear clinical significance and potential application value to develop a rapid, high detection sensitivity, and good reproducibility PCR detection kit for multiple detection of respiratory pathogen infection

Images