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

Abstract

The invention provides a molecular-biological detection method for chlamydia trachomatis 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 the 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 a target; step (2) adopting a RNAreal-time fluorescent constant-temperature amplification detection technology to obtain a detection result for chlamydia trachomatis 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 chlamydia trachomatis 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 Chlamydia trachomatis infection. Background technique [0002] Chlamydia is a prokaryotic microorganism that can pass through bacterial filters, has a unique developmental cycle, and is strictly intracellular. Chlamydial tyachomatis (CT) is the pathogen of various diseases such as trachoma, nongonococcal urethritis, cervicitis, pelvic inflammatory disease, endometritis, neonatal inclusion body conjunctivitis, infant pneumonia, etc. It is the most prevalent pathogen in the world. Causative agents of sexually transmitted diseases. The laboratory diagnosis of CT infection mainly includes cell biology methods (direct microscopy, isolation and culture), immunology methods (direct fluorescent antibody detection, enzyme-linked immunosorbent assay) and molecular b...

AI Technical Summary

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 Chlamydia trachomatis infection based on RNA target SAT. The combination of 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 application of the two technologies, but a specific target capture technology, which can also be called magnetic detection technology. The bead method obtains pure bacterial target nucleic acid (RNA) as the most important basis for accurate detection of Chlamydia trachomatis infection; the real-time fluorescent nucleic acid constant temperature amplification detection technology uses M-MLV reverse transcriptase, T7 RNA polymerase and optimized probe technology to achieve simultaneous , so as to overcome the complex operation, need to have a certain experience to do the detection, and time-consuming, manual operation, more complicated defects

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