Kit for rapid detection and genotyping of mycoplasma pneumoniae

Abstract

The invention provides a kit for rapid detection and genotyping of mycoplasma pneumoniae, belonging to the technical field of microbiological detection. The kit contains 2 specific primer pairs and 2 probes matched with the primer pairs, and the nucleotide sequences are shown by SEQ ID No.1-6 respectively; rapid detection and genotyping of mycoplasma pneumoniae are realized by a duplex fluorescent quantitative PCR (polymerase chain reaction) method, the detection sensitivity is 3CFU, the coincident rate between the genotyping accuracy and that of common genotyping methods reaches 100%, the genotyping sensitivity is much higher than that of common culture-dependent genotyping technologies, the genotyping time is greatly shortened, and the genotyping can be performed at the time of detection on mycoplasma pneumonia in a sample; and moreover, the shortcomings of low sensitivity, time and labor consuming and relatively high cost of the existing mycoplasma pneumoniae genotyping method are overcome, and the kit provided by the invention has good practical sample detection and genotyping ability and is suitable for popularization and application.

Description

technical field [0001] The invention relates to the technical field of microorganism detection, in particular to a kit for rapid detection and genotyping of Mycoplasma pneumoniae. Background technique [0002] Mycoplasma pneumoniae (Mycoplasma pneumoniae, Mp) is one of the important pathogens causing human respiratory tract infection, and about 10%-40% of community-acquired pneumonia is caused by its infection every year. Fluorescent PCR detection technology has gradually become the gold standard for the detection of Mycoplasma pneumoniae infection by virtue of its advantages of rapidity, high sensitivity and specificity, and has been widely used in clinical testing and scientific research. Genotyping Mycoplasma pneumoniae on the basis of detection is an important monitoring method for in-depth understanding of its antigenic variation and early warning and prediction of strain epidemic outbreaks. Studies have found that Mycoplasma pneumoniae can be divided into two types of...

AI Technical Summary

Problems solved by technology

[0003] Although the commonly used Mycoplasma pneumoniae typing techniques reported above have been widely used, they have obvious defects: these methods all rely on the isolation and culture of Mycoplasma pneumoniae, that is, after the Mycoplasma pneumoniae strains are isolated from the specimens, they are purified and purified. Genotyping can only be performed after culturing and extracting high-concentration Mycoplasma pneumoniae genomic DNA, and the amount of Mycoplasma pneumoniae nucleic acid in ordinary clinical specimens is not enough to support the detection by the above-mentioned typing method

This situation has led to a serious disconnection between the detection technology and the typing technology of Mycoplasma pneumoniae: after nucleic acid extraction, the rapid detection of Mycoplasma pneumoniae is carried out by fluorescent PCR technology (the test result can be obtained within 1 hour), but the typing needs to be determined first. The specimens were isolated and cultured for Mycoplasma pneumoniae strains, and the positive specimens could be tested by common typing methods after nucleic acid extraction (the typing results could be obtained after an average of 2 weeks), and the culture of Mycoplasma pneumoniae specimens was very difficult and the positive rate was low, resulting in Many positive specimens detected by fluorescent PCR cannot be isolated and cultured for strains and cannot be tested for typing, which makes the typing results biased to a certain extent

The inconsistency between the detection sensitivity of Mycoplasma pneumoniae and genotyping detection, and the asynchronous detection time limits have seriously restricted the development of Mycoplasma pneumoniae research.

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