Preparation method, amplification primers and detection reagent for HPV full-length genome quality control product

Abstract

The invention relates to a preparation method, amplification primers and detection reagent for a HPV full-length genome quality control product. The preparation method of the HPV full-length genome quality control product comprises: (1) acquiring a DNA template, (2) carrying out PCR amplification through amplification primers, (3) connecting a target DNA fragment to a vector by a HD Cloning linkage technology, (4) introducing the plasmid into a competent cell and then extracting the plasmid and (5) diluting the plasmid with a human genome. The preparation method of the human papillomavirus full-length genome quality control product can successfully acquire the HPV gene detection quality control product which has no risk of biological infection, can simulate a clinical sample, can be mass-produced, has a controllable concentration and can be stably preserved. The preparation method has good applicability.

Description

technical field [0001] The invention relates to gene detection technology, in particular to a preparation method of HPV full-length genome quality control product, amplification primers and detection reagents. Background technique [0002] Human papillomavirus (Human Papillomavirus, HPV) is a group of non-enveloped small DNA viruses, belonging to Papillomaviridae. The morphological feature of the HPV virus is usually an icosahedron with a diameter of 52-55 nm. The length of its genomic DNA is about 8000bp, and the genome is functionally divided into three parts: including early transcription regions (coding early proteins such as E1, E2, E4, E5, E6, E7, etc., involved in virus replication, transcription, translation regulation and transformation, etc. function), late transcribed region (encodes major capsid protein L1 and minor capsid protein L2 for virus assembly; L1 accounts for about 80% of the capsid protein and is highly conserved, L2 has less content and more variatio...

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Problems solved by technology

[0009] The national standard materials mentioned above are more expensive, the output is small, and the genotypes contained do not cover the common types of HPV39, 52, 68, etc., which cannot meet the standardization of reagents in my country, the performance evaluation of reagent methods in laboratories, indoor quality control and laboratory testing. The need for interim quality assessment

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