Primers, probe, detection method and kit for PCR-fluorescence detection of human papilloma virus types 16 and 18

Abstract

The invention discloses primers, a probe, a detection method and a kit for PCR-fluorescence detection of human papilloma virus types 16 and 18, and the detection method relates to a nucleic acid detection technology for detecting the high-risk human papilloma virus (HPV) types 16 and 18 which are the most common cause of cervical lesions. The kit comprises a PCR nucleic acid reaction mixture liquid, wherein the PCR nucleic acid reaction mixture liquid comprises sterile water, a dNTP mixture, a 10 * PCR buffer, a solution containing Mg<2+> ions, dUTP, forward primers of genes of the HPV types 16 and 18, reverse primers of the genes of the HPV types 16 and 18, and the probe for the genes of the HPV types 16 and 18. The PCR-fluorescence tube-combining detection of the HPV types 16 and 18 is carried out by using the locked nucleic acid (LNA) probe, and the primers, the probe, the detection method and the kit have the advantages of specificity, sensitivity, rapidity and simple operation.

Description

technical field [0001] The invention relates to the technical field of biological detection, and relates to a primer, a probe, a detection method and a kit for PCR-fluorescent detection of 16&18 human papillomaviruses, which are suitable for 16&18 of genitourinary tract, cervical exfoliated cells, tissues and other body fluids Human papillomavirus (HPV) nucleic acid detection. Background technique [0002] More than 100 genes of HPV (human papillomavirus) have been identified in the past 20 years, and at least 85 genes have been completely sequenced. HPV belongs to the papillomavirus genus of the Papovaviridae family. It is a double-stranded circular DNA virus of about 7200-8000 bp. The genome encodes three functional regions, namely, the early transcription region, the late transcription region and the upstream regulatory region (URR). The early region accounts for about 4kb, encoding 5 to 8 open reading frames, which are E6, E7, E1 (E8), E2, E4 (E3), and E5, and are invol...

AI Technical Summary

Problems solved by technology

Advantages of this method: simple operation, low price, suitable for preliminary screening; disadvantages: hollow cells are the main morphological changes of HPV infection, but due to other viral infections and human factors, vacuoles or hollows may appear in the cells In addition, cytological examination is easily affected by factors such as sampling, staining, and subjective judgment of cytopathologists. Therefore, the application of cytopathological detection of HPV has low sensitivity, poor specificity, high false negative rate and false positive rate, and cannot detect HPV. type

Advantages of this method: simple operation, low price, suitable for preliminary screening; disadvantages: low accuracy, high quality requirements for personnel, and great pain for patients

Advantages of this method: clear principle and relatively simple operation; disadvantages: the objects of serological detection are antigens and antibodies. Since the human body has a certain hysteresis in the immune response to HPV, serological detection is not suitable for those with no immune response and HPV latent infection. Will produce missed detection

However, the traditional 16 and 18 human papillomavirus fluorescent probe detection kits design primers and probes for 16 and 18 types of human papillomavirus respectively, so as to separate the detection, the method is cumbersome, and lacks the significance of clinical rapid and simple detection

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