RT-PCR method for overspeed detection of trichomonas vaginalis

Abstract

The invention relates to an RT-PCR (Reverse Transcription-Polymerase Chain Reaction) method for ultrafast detection of trichomonas vaginalis, which comprises a reaction procedure for designing a pair of specific primers and probes PF1, PR1 and FQQ aiming at 18S ribosome RNA (Ribose Nucleic Acid) sequence of trichomonas vaginalis and carrying out real-time fluorescent quantitative PCR detection on nucleic acid of a sample to be detected by adopting a fluorescent quantitative PCR reaction. The real-time fluorescent quantitative reverse transcription PCR reaction procedure comprises the following steps: carrying out pollution digestion at 37 DEG C for 2 minutes; pre-denaturation is performed at 95 DEG C for 30 s; denaturation is conducted at 95 DEG C for 2 s, annealing extension is conducted at 69 DEG C for 13 s, and 45 cycles are conducted. According to the primer and the probe disclosed by the invention, the extension temperature meets the optimal extension temperature of Taq DNA polymerase, so that the amplification time is effectively shortened. The extension primer is completely matched with a template, and the Tm value reaches about 69 DEG C, so that annealing and extension can be carried out at the same time at 69 DEG C, two-step amplification can be directly adopted from the beginning, the heating and cooling processes are reduced, and the detection time is effectively shortened.

Description

technical field [0001] The invention relates to the field of molecular biology detection technology and molecular diagnosis, in particular to an RT-PCR method for ultra-fast detection of Trichomonas vaginalis. Background technique [0002] Vaginitis, also known as vaginitis, is a group of conditions that cause vulvovaginal symptoms such as itching, burning, irritation, and abnormal discharge. Vaginitis is one of the most common infectious diseases in the female reproductive system. According to the epidemiological survey, more than 50% of the total surveyed people have genital infection, and the infection of vaginitis ranks first among the infectious diseases of the female reproductive system. The serious development of the disease can lead to cervicitis, pelvic inflammatory disease, endometritis Wait. More importantly, it will lead to a series of adverse outcomes such as infertility, miscarriage, and infection of the fetus, and even further increase the risk of cervical c...

AI Technical Summary

Problems solved by technology

The conventional method is to set the annealing extension temperature of Taq DNA polymerase to the annealing temperature of the probe -5°C (about 60°C), but the existing method has the following disadvantages: Taq DNA polymerase does not reach the highest activity during the working process State, so the amplification rate does not reach the highest level, resulting in a longer detection time

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