RNA (Ribonucleic Acid) target SAT (Simultaneous Amplification and Testing) based molecular biological detection method for ureaplasma urealyticum infection

Abstract

The invention provides an RNA (Ribonucleic Acid) target SAT (Simultaneous Amplification and Testing) based molecular biological detection method for ureaplasma urealyticum infection. The method comprises the following steps: I, by using a specific target capturing technique, extracting pure cell target nucleic acid by using a magnetic bead method, that is, taking RNA as a target, namely (1-1) manually extracting target nucleic acid specificity, and (1-2) extracting pure cell target nucleic acid, that is, taking RNA as the target; II, obtaining detection results of chlamydia trachomatis infection by using an RNA real-time fluorescent constant-temperature amplification detection technique, namely (2-1) generating one DNA (Deoxyribonucleic Acid) copy of the target nucleic acid RNA through M-MLV reverse transcriptase, (2-2) generating multiple RNA copies from the DNA copy by using T7RNA polymerase, and (2-3) specifically combining an optimized probe with a fluorescent marker and the extracted RNA copies so as to generate a fluorescent signal which is captured by using a detector, and obtaining a detection result of chlamydia trachomatis infection according to signal appearance time andintensity; III, with the combination of a positive reference and a negative reference, judging detection results, and carrying out bacterium testing.

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 Ureaplasma urealyticum infection. Background technique [0002] Ureaplasma urealyticum is a kind of prokaryotic microorganism, whose size is between bacteria and viruses. It is a common pathogen of human genitourinary tract. It is related to many urogenital tract infection symptoms, perinatal infection and infertility. It is one of the pathogens of sexually transmitted diseases. It is worth noting that among pregnant women, there are more cases of Ureaplasma urealyticum infection, 80% reported abroad, and 55.12% reported in my country. Infection with Ureaplasma urealyticum during this period not only endangers the health of the mother, but also endangers the survival of the fetus. It is easy to cause low birth weight infants, neonatal respiratory tract inf...

AI Technical Summary

Problems solved by technology

[0012] 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 Ureaplasma urealyticum infection based on RNA target SAT. Moreover, the combination with the fluorescent constant temperature amplification detection technology (SAT) organically combines the advantages of RNA targets 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 be called the magnetic bead method to obtain pure bacterial target nucleic acid (RNA) as the most important basis for accurate detection of Ureaplasma urealyticum infection; real-time fluorescent nucleic acid constant temperature amplification detection technology uses M-MLV reverse transcriptase, T7 RNA polymerase Simultaneous implementation with optimized probe technology, thus overcoming the complex operation, requiring a certain amount of experience to do a good job of detection, and time-consuming, manual operation, and more complicated defects

Images