Unlock instant, AI-driven research and patent intelligence for your innovation.

HPV extracting method of cervical exfoliated cells

A technology for exfoliating cells and cervical epithelial cells, applied in the directions of microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve the problems of false negative HPV hybridization results, poor DNA quality, and ineffective removal of mucus and hemoglobin. , to achieve the effect of simplifying experimental steps and good typing effect

Inactive Publication Date: 2016-07-06
李达
View PDF3 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For such samples, if the traditional DNA separation method is used to extract HPV DNA, the mucus and hemoglobin are not effectively removed, which may easily lead to poor quality of the extracted DNA, PCR reaction inhibitors and false negative results of HPV hybridization.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • HPV extracting method of cervical exfoliated cells
  • HPV extracting method of cervical exfoliated cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] A method for extracting HPV from exfoliated cells of the cervix, comprising the steps of:

[0022] (1) Separation of cervical epithelial cells: take 500 μl of cervical fluid and centrifuge at a rate of 13,000 r / min for 4 minutes, and discard the supernatant to obtain centrifugal cervical fluid;

[0023] (2) HPV release: directly add 400 μl of solution I and 25 μl of proteinase K to the centrifuged cervical fluid obtained in step (1), oscillate and mix, and place in an incubator at 56-60° C. for 12-15 hours;

[0024] (3) Separation of HPV: draw 500 μl of supernatant, add 400 μl of solution II;

[0025] (4) HPV treatment: oscillate and mix, let stand for 2min, centrifuge at 13000r / min for 7-9min, aspirate and discard the supernatant, jog centrifugation, and then aspirate and discard the remaining supernatant; add 60 μl solution III, and let stand for 10- 12min, centrifuge at 13000r / min for 1-2min;

[0026] (5) Detection of HPV purity, content identification and positive...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention discloses a method for extracting HPV from cervical exfoliated cells, which comprises the following steps: (1) separation of cervical epithelial cells, (2) release of HPV, (3) separation of HPV, (4) treatment of HPV, (5) For the detection of HPV purity, content identification and positive rate, by using proteinase K to digest hemoglobin and mucus protein, PCR reaction inhibitors can be effectively removed, and the experimental steps are simplified, and the HPV typing effect is very good.

Description

technical field [0001] The invention relates to the technical field of gynecological medicine, in particular to a method for extracting HPV from exfoliated cells of the cervix. Background technique [0002] HPV is a double-stranded closed circular small DNA virus with strict host range and tissue specificity, which can cause benign and malignant tumors of human epithelium. According to different HPV gene sequences, more than 120 subtypes have been isolated, at least 40 of which are related to cervical cancer [1]. At present, HPV DNA typing detection, as an important indicator for clinical prediction of disease risk, selection of treatment options and HPV vaccine development research, has been paid more and more attention. The research on the HPV DNA extraction method is the basis and premise of its genotype detection, and the quality of the extracted DNA also directly affects the subsequent experimental operation. [0003] The traditional separation method used to extract ...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/00C12R1/93
CPCC12N7/00C12N2710/20051
Inventor 李娜柳燕于英英隋光丽陈丽媛王芳尹喜玲韩景奇
Owner 李达