Method for performing rapid gene typing on trace human papilloma virus (HPV)

A genotyping method and rapid technology, applied in biochemical equipment and methods, microbial determination/inspection, etc., can solve the problems of inability to distinguish specifically, low detection positive rate, high cost, and solve the problem of small detection range and shortened detection. Extraction time, the effect of improving specificity

Inactive Publication Date: 2010-09-22
SUN YAT SEN UNIV
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Problems solved by technology

[0005] At present, the detection methods of HPV mainly include: PCR detection, the most commonly used is MY09/11, GP05/06 primer amplification, and then combined with ELESA agent PCR reverse dot hybridization, but the follow-up work is complicated and requires high experimental technology; combined target Automatic gene detection, rapid typing of HPV by using gene chip and electric sensor counting, but the cost is high; histopathology combined with in situ hybridization technology, but needs to analyze the tissue, which is not suitable for early screening work; hybrid capture DNA analysis, but only limited 13 kinds of HPV can be detected, and HPV typing cannot be carried out; HPV DNA detection in blood circulation, its disadvantage is that the positive rate of detection is not high, which is about 20%, and it is not high in women with current infection Less than half of them c

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  • Method for performing rapid gene typing on trace human papilloma virus (HPV)
  • Method for performing rapid gene typing on trace human papilloma virus (HPV)
  • Method for performing rapid gene typing on trace human papilloma virus (HPV)

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Embodiment Construction

[0032] The present invention will be further elaborated below with regard to specific embodiment:

[0033] 1. Genome extraction:

[0034] The method is applied to the early screening of cervical cancer and has a wide range of applications, and can be used to detect the genome extraction of various types of samples such as paraffin sections, formalin-soaked tissues, blood samples, and exfoliated cells.

[0035] 1. Formalin-fixed tissue block genome preparation:

[0036] A. Take 0.01-0.05g tissue, wash it three times with phosphate buffer; B. Grind it in liquid nitrogen, add 600μL lysate (lysate formula: 10mmol / L Tris-HCl; 0.1mol / L EDTA; 0.5% (M / L V) SDS; 20ug / mg DNase-free pancreatic RNase; PBS solution formula: 8g NaCl, 0.2g KCl; 1.44g Na 2 HPO 4 ;0.24g KH 2 PO 4 , dissolved in 800mL water, HCl to adjust the pH to 7.4, add water to make up to 1L) "Molecular Cloning Second Edition" and 4μL proteinase K (20mg / mL), digest overnight at 55°C; C. 600uL saturated phenol extracti...

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Abstract

The invention discloses a method for performing rapid gene typing on trace human papilloma virus (HPV), which has the characteristics of wide application range, high sensitivity, accuracy and high efficiency. Based on a polymerase chain reaction (PCR) detection method, nested amplification is performed on an HPV L1 conservative region for three times, electrophoresis detection and sequencing are performed on a target product and the target product is typed by using special analysis software so as to correctly and rapidly diagnose trace HPV virus infection and detect and identify various HPV gene types, such as high-risk type, low-risk type and the like. The method has high sensitivity and wide application range, can be used for detecting various types of samples, such as paraffin sections, formalin-dipped tissues, blood samples, cast-off cells and the like, and is convenient to apply to early screening of cervical cancer.

Description

technical field [0001] The invention relates to a rapid genotyping method for trace HPV. Background technique [0002] Cervical cancer is the malignant tumor with the highest morbidity and mortality rate in the female reproductive tract. About hundreds of thousands of women die from cervical cancer in the world every year. It is the most serious disease that threatens women's lives. The occurrence and development of cervical cancer is a process from gradual change to mutation. Planned screening is very critical for effective prevention and early treatment of cervical cancer and its precancerous lesions. It is of great social significance to promote the application of advanced detection technology for early diagnosis and treatment of cervical cancer and precancerous lesions and to protect the reproductive health of women. [0003] Clinical scientific research shows that persistent infection of human papillomavirus (HPV) is the direct cause of cervical cancer. HPV can be div...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68
Inventor 吴玉萍常利红宁曦
Owner SUN YAT SEN UNIV
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