Nucleic acid detection kit for human papillomavirus and method and application thereof

Abstract

The invention discloses a nucleic acid detection kit for human papilloma virus, a use method and an application thereof. The kit includes a nucleic acid amplification reagent comprising a primer pair and a probe corresponding to the primer pair. The use method of the kit includes the following steps: 1) extracting nucleic acids from a sample; 2) preparing the reagent; 3) performing PCR amplification; and 4) performing fluorescent detection to a PCR amplification reaction product at 65-72 DEG C, and determining infection type of the HPV according to the changes on the Ct value of a target amplification curve and the Ct value of an internal standard amplification curve. The kit can be used for detecting low-risk and high-risk human papilloma virus and for typing the human papilloma virus. The kit can be used for calculating relative viral load of the HPV, can avoid pollution, can increase treatment throughput of samples, and can avoid missing detection.

Description

technical field [0001] The invention relates to a nucleic acid detection kit and its use method and application, in particular to a human papillomavirus nucleic acid detection kit and its use method and application. Background technique [0002] Human papillomavirus (HPV) is a kind of papillomavacuolating virus A belonging to Papovaviridae. It is a spherical DNA virus with high specificity. It can infect the squamous epithelium of skin and mucous membrane and cause proliferation. At present, more than 130 kinds have been isolated, and different types cause different clinical manifestations. According to the different tissue parts invaded, they can be divided into: [0003] Skin low-risk types (HPV1, 2, 3, 4, 7, 10, 12, 15, etc.), which are related to common warts, flat warts, plantar warts, etc.; [0004] High-risk skin types (HPV5, 8, 14, 17, 20, 36, 38); [0005] Mucosa low-risk types (HPV6, 11, 13, 32, 34, 40, 42, 43, 44, 53, 54, etc.); [0006] Mucosa high-risk types ...

AI Technical Summary

Problems solved by technology

[0012] 3) Second-generation hybrid capture, HCII: the first FDA-approved method for clinical testing applications, which can distinguish between high and low risks, and can be relatively quantified. The disadvantages are: no internal standard, long detection time (DNA decomposition 45min, DNA-RNA hybridization 60 minutes, antibody capture 60 minutes, chemiluminescence 30 minutes, computer interpretation 15 minutes), low sensitivity, high cost, and cross-reaction with HPV subtypes outside the detection range, resulting in false positive results

Disadvantages: low sensitivity, easy to pollute when opening the cover, expensive

Advantages: Can be classified into specific types; Disadvantages: Manual operation is cumbersome, easy to pollute when opening the cover, and expensive

However, the fluorescent PCR method is limited by the detection channel. Generally, only 16, 18, 6, and 11 can be typed, and other types cannot be specifically typed.

At the same time, the target sequence of most PCR typing products is the L region, which cannot accurately distinguish some types

[0016] In summary, cervical cytology detection and pathological examination are greatly affected by subjective factors, and the sensitivity is low, so there is a certain rate of missed detection in clinical application; nucleic acid hybridization method, hybrid capture method, molecular diversion hybridization method, Compared with the former, the liquid chip method has improved sensitivity and a larger coverage for HPV type detection, but it is prone to contamination during operation, resulting in false positive results; real-time fluorescent PCR method has higher sensitivity and specificity. It can greatly reduce the false positive rate and missed detection rate, and the detection process can be fully automated, reducing human input. However, due to the restriction of the number of fluorescent channels and the difficulty of multiplex PCR technology, there are certain limitations in determining the HPV type. limit

[0017] The above-mentioned detection methods all have their own defects, which bring certain difficulties to the diagnosis and treatment of clinical HPV.

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