Method using loop-mediated isothermal amplification technology to detect mycoplasma pneumoniae

Abstract

The invention discloses an LAMP (loop-mediated isothermal amplification) detection method of mycoplasma pneumoniae and the special primers and kit of the LAMP detection method. The special primers are used for performing fast isothermal amplification on the mycoplasma pneumoniae. The LAMP primers are designed according to the specific conservative sequence of pneumonia, and each group of primers comprises 4 oligonucleotides. When the special primers are used for mycoplasma pneumoniae detection, the primers are in the form of white precipitate under naked-eye observation; after SYBR GREEN is added during positive reaction, fluorescent green is evidently enhanced under ultraviolet-lamp observation. A real-time turbidity detection result shows that product turbidity increases along with the prolonging of reaction time, and the primers are in the form of trapezoid strips after gel electrophoresis detection. The method has the advantages that a new technical platform is provided for the mycoplasma pneumoniae detection, to-be-detected samples can be DNA extracted by commercial kits or purified DNA and coarsely-extracted DNA extracted by methods represented by a boiling method, and the method is suitable for being popularized and applied in basic units, field monitoring and bedside detection.

Description

technical field [0001] The invention belongs to the application of a molecular biology detection method represented by isothermal amplification in the detection of mycoplasma pneumoniae, that is, a loop-mediated isothermal amplification detection technology of mycoplasma pneumoniae and its special primers, detection method and kit. Background technique [0002] Mycoplasma (Mycoplasma) is a prokaryotic cell-type microorganism between bacteria and viruses. So far, there are about 20 species of mycoplasma that can cause human infection, among which mycoplasma pneumoniae (M.pneumoniae, Mp) is the most common. Mp is one of the important pathogens causing human respiratory tract infection, about 10% to 30% of community-acquired pneumonia is caused by its infection every year. Mp infection is distributed globally, it can be transmitted through droplets in the form of aerosol particles, and the population is generally susceptible. When Mp invades the respiratory tract, it locates ...

AI Technical Summary

Problems solved by technology

[0003] At present, the gold standard for laboratory diagnosis of mycoplasma is isolation culture, but the isolation culture method usually has two disadvantages. One is that the detection time is too long. It takes at least 3 weeks for a typical colony to grow on a solid plate, which is of little significance for rapid clinical diagnosis.

Second, isolation and culture are difficult and demanding, and it is difficult to promote in general hospitals and laboratories

[0005] The application of LAMP in the detection of Mycoplasma pneumoniae has been reported at home and abroad, but so far, there are no LAMP-specific primers and kits for the detection of Mp on the market.