Process for detecting and typing all known genital tract human papillomavirus

A technology of human papillomavirus and reproductive tract, which is applied in the direction of biochemical equipment and methods, and the determination/testing of microorganisms, can solve the problems of non-typing, low sensitivity of the method, and high technical cost, and achieve simple and convenient operation, Low cost, low cost effect

Inactive Publication Date: 2006-08-30
ATTACHED OBSTETRICS & GYNECOLOGY OSPITAL MEDICALCOLLEGE ZHEJIANG UNIV
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Problems solved by technology

Since different reproductive tract mucosal HPV genotypes have different biological characteristics and different risks, it is necessary to detect and type HPV at the same time; at the same time, with the development of vaccine research, a good HPV typing method should be established It is very important for the targeted use of vaccines, but so far, there is no suitable method to detect and type all HPVs that are currently known to infect the genital mucosa
In recent years, there have been many studies on papillomavirus typing, but some methods have low sensitivity, such as Southern blotting and in situ hybridizatio

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  • Process for detecting and typing all known genital tract human papillomavirus

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Embodiment

[0026] 1. Extract DNA from cervical secretions of 200 patients with cervical cancer and cervical intraepithelial neoplasia

[0027] 1) Take the cervical secretions with a cotton swab, shake it in 1ml of normal saline for a few minutes, discard the cotton swab, and save the liquid as the specimen;

[0028] 2) Take 500 μl of the specimen, add 200 μl of lysate (containing 40 μg of proteinase K), and incubate at 55°C for 30-60 minutes;

[0029] 3) Add an equal amount of 1:100 phenol chloroform, vibrate vigorously for 15 seconds, and let stand on ice for 5 minutes;

[0030] 4) Centrifuge at 12000rpm at 4°C for 10min, and take the supernatant;

[0031] 5) Add 2 times of absolute ethanol and let stand on ice for 30-60min;

[0032] 6) Centrifuge at 12000rpm 4°C for 30min, discard the supernatant;

[0033] 7) Add 1000ml of 70% ethanol, centrifuge at 12000rpm at 4°C for 10min, and discard the supernatant;

[0034] 8) Repeat step 6;

[0035] 9) Dry in air, add 100 μl TE (PH=8.0), an...

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Abstract

This invention discloses a method of quickly distinguish and classify existing genital meatus human papilloma virus. The method can distinguish existing 48 type HPV that can infect genital meatus mucous membrane and its normal mixed infection. And it can select Rsa I,Mse I,Pst I and HaeIII four normal using inscribing enzyme combinations according to optimal selection, optimize assort and at the same time the sensitivity and specificity can be ensured. Then HPV gene type is judged according to the established genital HPV gene type MY09/11 amplification product RMPH enzyme inscribing data. The operation is simple and time can be saved, the cost is low, its equipment needed exists in county hospital, and its cost is very low. It can be high pass quantity experienced, the technical operability of developing HPV inflection epidemiology investigation.

Description

technical field [0001] The invention relates to a method for detecting human papillomavirus of reproductive tract, in particular to a PCR-RFLP (polymerase chain reaction-restriction fragment length polymorphism analysis) capable of detecting and typing almost all known human papillomaviruses of reproductive tract )method. Background technique [0002] Cervical cancer is the most common reproductive tract malignancy in women, and it has been confirmed that human papillomavirus (HPV) is the key pathogenic factor of cervical cancer and its precursor lesions. More than 200 types of HPV genotypes have been found, of which about 48 types are related to genital tract mucosal lesions. Since different reproductive tract mucosal HPV genotypes have different biological characteristics and different risks, it is necessary to detect and type HPV at the same time; at the same time, with the development of vaccine research, a good HPV typing method should be established It is very import...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 陈怀增叶枫谢幸吕卫国洪蝶梁朝霞
Owner ATTACHED OBSTETRICS & GYNECOLOGY OSPITAL MEDICALCOLLEGE ZHEJIANG UNIV
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