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Method of detecting human papilloma virus 16/18 type gene in urine by using digital PCR (polymerase chain reaction)

A human papillomavirus, digital technology, applied in the field of cervical cancer detection, can solve the problems of inflammation risk and poor detection accuracy, and achieve the effect of making up for the difficulty of detection

Inactive Publication Date: 2017-05-31
安徽安龙基因科技有限公司
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AI Technical Summary

Problems solved by technology

[0006] The purpose of the present invention is to solve the defects of poor detection accuracy and risk of inflammation in the prior art, and provide a method for detecting human papillomavirus 16 / 18 gene in urine by digital PCR to solve the above problems

Method used

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  • Method of detecting human papilloma virus 16/18 type gene in urine by using digital PCR (polymerase chain reaction)

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0076] Taking the detection of HPV16 and 18 E7 genes as an example

[0077] The specific steps are:

[0078] Step 1. Extraction of DNA in urine:

[0079] a. Urine specimen collection: Collect 30 mL of the first morning urine from patients with cervical cancer and precancerous lesions, put it into a sterile test tube with a cover, centrifuge at 5000rpm for 10min at 4°C, and collect the sediment at the bottom;

[0080] b. Add 1.0ml TE buffer solution to the precipitate collected in step a, mix well, centrifuge at 8000rpm for 5min, and collect the precipitate at the bottom;

[0081] c. Add 1.0ml TE buffer solution to the precipitate collected in step b, mix well, centrifuge at 8000rpm for 5min, and collect the precipitate at the bottom;

[0082] d. Add 450ul of lysate and 10ul of proteinase K to the precipitate collected in step c, and put it in a water bath for 1 hour at 55°C to dissolve the precipitate to obtain a mixed solution;

[0083] e. Take out the mixed solution, add ...

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Abstract

The invention discloses a method of detecting a human papilloma virus 16 / 18 type gene in urine by using digital PCR (polymerase chain reaction). The method comprises the particular steps of extracting DNA (deoxyribonucleic acid) from the urine, preparing a primer reaction solution, preparing a droplet, performing PCR amplification, detecting the droplet, analyzing data and displaying a result, wherein a lysate comprises 10mM of Tris-HCl, 1mM of EDTA (ethylene diamine tetraacetic acid) and 0.5% of SDS (sodium dodecyl sulfonate); a pH (potential of hydrogen) value of a sodium acetate solution is 5.2. The detection mode is free from age limit and available; sampling is noninvasive; the method has high sensitivity and high precision and overcomes the detection difficulty arising from little viral nucleic acid in the urine; HR-HPV detection in the urine can be widely applied to clinical diagnosis and treatment and cervical cancer screening; the method has the advantage that the method has an important significance for prevention and treatment of a cervical cancer.

Description

technical field [0001] The invention relates to the technical field of cervical cancer detection, in particular to a method for detecting human papillomavirus 16 / 18 gene in urine by using digital PCR. Background technique [0002] Cervical cancer is a malignant gynecological tumor with a high incidence rate. Among female patients with malignant tumors worldwide, the number of cervical cancer patients is only lower than that of breast cancer patients, which seriously affects the health and quality of life of patients. , HPV is a small molecule double-stranded DNA virus that mainly infects human mucous membranes and skin squamous epithelial tissues. Human papillomavirus (high risk human papillomavirus, HR-HPV) is a necessary factor to cause cervical cancer and precancerous lesions. HR-HPV infection and detection have become one of the key issues in screening and preventing cervical cancer. [0003] At present, clinically commonly used cervical cancer detection methods mainly ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68C12R1/93
CPCC12Q1/6851C12Q1/708C12Q2531/113C12Q2561/101C12Q2563/159
Inventor 韦玉军李航李静苏军吴远航
Owner 安徽安龙基因科技有限公司
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