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Method for detecting HPV and application of Phi29DNA polymerase in detecting HPV

A polymerase and virus detection technology, which is applied in biochemical equipment and methods, microbial measurement/inspection, etc., can solve the problem of low HPV detection rate, achieve stable amplification output, strong clinical use value, and simple operation Effect

Inactive Publication Date: 2010-01-06
SHANGHAI INST OF IMMUNOLOGY +1
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Problems solved by technology

[0017] In order to solve the problem that the detection rate of HPV is too low in the prior art, the present invention provides a method for HPV detection. The method is to firstly use Phi29 DNA polymerase to perform multiple replacement amplification of the whole gene on the patient sample, and then use diversion hybridization detection, thereby greatly improving the detection rate of HPV

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  • Method for detecting HPV and application of Phi29DNA polymerase in detecting HPV
  • Method for detecting HPV and application of Phi29DNA polymerase in detecting HPV
  • Method for detecting HPV and application of Phi29DNA polymerase in detecting HPV

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Embodiment 1

[0037] Materials and Methods

[0038] Materials: 223 samples of gynecological outpatients whose test results were negative by the Cape HPV diversion hybridization instrument. The samples came from the following hospitals in Shanghai: Shanghai Sixth People's Hospital, Shanghai Ninth People's Hospital, Shanghai Putuo District Women and Children Health Center, Huangpu District Maternal and Child Health Hospital, Guobin Medical Center, and Armed Police Hospital are all outpatients; Phi29 enzyme was purchased from NEB Company (M0269, NewEngland Biolabs) in the United Kingdom; Kaipu diversion hybridization instrument was purchased from China Kaipu Biotechnology Co., Ltd. .

[0039] Extraction of DNA from patient samples: using the kit provided by Hybribio (Hybribio, HPV Genotyping Detection Kit).

[0040] Whole-genome Multiple Displacement Amplification (MDA): Operate according to NEB instructions, and the specific steps are as follows: Take 1 μL of reaction buffer, 2.5 μL of rando...

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Abstract

The invention discloses a method for detecting human papillomavirus (HPV) in a sample, which sequentially comprises the following steps: A) extracting DNA in the sample; B) amplifying the extracted DNA in step A); C) amplifying PCR; and D) detecting viruses. The amplifying of the step B) is to carry out multiple displacement amplification on all the genes of the extracted DNA with a Phi29DNA polymerase. The invention also discloses application of the Phi29DNA polymerase in detecting the human papillomavirus. The detection method can obviously improve the detection rate of the HPV. The sample does not need to be purified when the Phi29DNA polymerase is used to amplify the DNA. A gene group is uniformly amplified; the amplifying yield is stable; the operation is simple; the method is independent of a PCR reaction; and no amplified product having specificity is generated.

Description

technical field [0001] The invention belongs to the technical field of virus detection, in particular to a method for detecting HPV and the application of other Phi29 DNA polymerases in virus detection. Background technique [0002] Cervical cancer is the second highest cause of cancer death among women in the world. There are about 450,000 new cases every year in the world, and the mortality rate is 50%. Drs.Rous and Beard first proposed that HPV is a carcinogen in 1934. Since Zur Hausen proposed that HPV is the most likely factor of cervical cancer in 1974, scholars at home and abroad have conducted a lot of research on the relationship between HPV infection and cervical cancer. And obtained a lot of laboratory and clinical evidence. In 1977, Laverty observed the presence of HPV particles in cervical cancer biopsy tissue under an electron microscope. In 1981, HPV was isolated from the reproductive tract for the first time. From 1983 to 1986, HPV16, 18, 31, and 35 were iso...

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68
Inventor 张艳管爱民顾关林马式薇何昕李宁丽黄立东姜惠明王利沈佰华蔡佩明陈勇
Owner SHANGHAI INST OF IMMUNOLOGY
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