Molecular-biological detection method for neisseria gonorrhoeae infection based on SAT (Simultaneous Amplification and Testing) of RNA target

Abstract

The invention provides a molecular-biological detection method for neisseria gonorrhoeae infection based on SAT (Simultaneous Amplification and Testing) of a RNA target. The molecular-biological detection method comprises the following steps: step (1) adopting a specific target capture technology and a magnetic-bead method to extract pure cell target nucleic acid, i.e., adopting RNA as a target, wherein comprising (1-1) the specificity of the target nucleic acid is manually extracted; (1-2) the pure cell target nucleic acid is obtained, i.e., RNA is used as the target; step (2) adopting a RNAreal-time fluorescent constant-temperature amplification detection technology to obtain a detection result for neisseria gonorrhoeae infection, wherein comprising (2-1) M-MLV reverse transcriptase generates a DNA copy of the target nucleic acid; (2-2) T7RNA polymerase generates multiple RNA copies from the DNA copy; (2-3) an optimizing probe with fluorescence labeling is specifically combined withthe extracted RNA copy, so that a fluorescence signal generated is captured by a detection instrument, and the detection result for the neisseria gonorrhoeae infection is obtained according to the occurring time and the intensity of the signal; step (3) judging the detection result by combination with positive control and negative control.

Description

technical field [0001] The invention relates to the technical field of molecular biology detection and identification of bacterial infection, in particular to a method for molecular biology detection and identification of Neisseria gonorrhoeae infection. Background technique [0002] Gonorrhea is an infection caused by Neisseria gonorrhoeae. It is one of the main types of sexually transmitted diseases. It has a short incubation period and is highly contagious. The most common symptomatic infection of the urogenital system, such as urethritis, cervicitis, Epididymitis, prostatitis, salpingitis, etc. If not treated in time, gonococcal bacteria can invade the eyes, pharynx, skin, rectum, pelvic cavity and other parts, and even spread to other parts through the blood stream, forming gonococcal arthritis and tenosynovitis , meningitis, sepsis, etc. On the other hand, 5%-20% of male and 60% of female patients present asymptomatic infection. [0003] At present, the laboratory de...

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Problems solved by technology

[0010] In order to overcome the above defects in the detection methods of the prior art, the present application provides a molecular biology detection method for Neisseria gonorrhoeae infection based on RNA target SAT, the inventive concept of which is to obtain pure RNA targets through specific target capture technology, and The combination with fluorescence constant temperature amplification detection technology (SAT) organically combines the advantages of RNA target and SAT technology. Of course, the technical solution of the present invention is not a simple superposition of the two technologies, but through specific target capture technology, it can also be The pure bacterial target nucleic acid (RNA) obtained by the magnetic bead method is the most important basis for accurate detection of Neisseria gonorrhoeae infection; the real-time fluorescent nucleic acid constant temperature amplification detection technology uses M-MLV reverse transcriptase, T7 RNA polymerase and optimized Probe technology is realized at the same time, so as to overcome the complex operation, need certain experience to do the detection well, and time-consuming, manual operation, and relatively complicated defects

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