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

A technique for amplifying primers and whole genomes, which is applied in biochemical equipment and methods, microbe determination/inspection, recombinant DNA technology, etc. It can solve the problems that cannot meet the performance evaluation of reagent methods, internal quality control, external quality evaluation, and low output. , more expensive and other issues, to achieve the effect of avoiding low connection efficiency, good applicability, and controllable concentration

Active Publication Date: 2018-12-04
亚能生物技术(深圳)有限公司
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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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  • Preparation method, amplification primers and detection reagent for HPV full-length genome quality control product
  • Preparation method, amplification primers and detection reagent for HPV full-length genome quality control product
  • Preparation method, amplification primers and detection reagent for HPV full-length genome quality control product

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Embodiment

[0110] see Figure 1-8 , Figure 1-2 Electrophoresis results for electrophoresis of PCR products after sample DNA amplification; image 3 It is a schematic diagram of the structure of TVector (including the cloning site); Figure 4 Select the fluorescence profile for the sample, Figure 4 The middle curve corresponds to standard 1, sample, standard 2, standard 3 and standard 4 from left to right; Figure 5 is the standard curve formula; Image 6 The initial Ct value map for the stability study; Figure 7 Ct value change chart for stability research; Figure 8 It is the chart of the value determination results of 23 quality control substances by digital PCR absolute quantification method.

[0111] The preparation method of the human papillomavirus full-length genome quality control product of the present embodiment comprises:

[0112] (1) HPV nucleic acid extraction and type determination

[0113] Use the nucleic acid extraction reagent of Yaneng Biotechnology (Shenzhen)...

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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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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12Q1/6806C12Q1/70C12Q1/6851C12N15/70
CPCC12N15/11C12N15/70C12Q1/6806C12Q1/6851C12Q1/708C12Q2531/113C12Q2563/107
Inventor 卢舟宇田洁陈永娟陈志强蔡泽加
Owner 亚能生物技术(深圳)有限公司
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