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Cervical cell preservation and DNA fast extraction integrated kit and extraction method

A cervical cell and kit technology, which is applied in the field of molecular biology, can solve the problems of low DNA yield, health hazards of operators, and decreased sensitivity, and achieves the effects of low instrument dependence, high DNA purity, and guaranteed accuracy.

Active Publication Date: 2016-03-02
GWP BIOTECHNOLOGIES INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, methanol, formaldehyde and other fixatives are added in liquid-based cytology, which are highly destructive to DNA and have a serious impact on later PCR and other tests.
Moreover, both formaldehyde and methanol have certain toxicity, and they also have certain hazards to the health of operators.
In addition, most of the preservation solutions currently need to be transported under the protection of dry ice or ice boxes, which will undoubtedly increase the transportation cost and workload
[0004] At present, DNA extraction using DNA extraction kits is generally divided into several steps such as lysis, washing, and elution. The most critical step for DNA yield is the lysis step. If the lysis time is short, the DNA yield will be too high. Low, so that the sensitivity of later detection is reduced. At present, the cell sample is usually placed in the lysate and incubated at a certain temperature for 10-15 minutes. The cleavage step is cumbersome,

Method used

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  • Cervical cell preservation and DNA fast extraction integrated kit and extraction method
  • Cervical cell preservation and DNA fast extraction integrated kit and extraction method
  • Cervical cell preservation and DNA fast extraction integrated kit and extraction method

Examples

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example 1

[0034] Using the results of cervical cell sampling in a maternal and child health care hospital, the cervical cells collected by the cervical brush were placed in 1ml of cell preservation solution and mixed evenly to obtain a cell sample solution, wherein the components of the cell preservation solution were: 2mol / L guanidine hydrochloride, 5mmol / L Tris-HCl, 5mmol / L EDTA and 5v / v% Trixon-100, the rest is deionized water. After the sample was collected and transported at room temperature for 2 days, a sample was randomly selected for extraction. The components of the extraction reagent were as follows:

[0035] Wherein the first washing liquid contains the EDTA of the ethanol of 50v / v%, the guanidine thiocyanate of 1mol / L, the Tris-HCl of 10mmol / L and the 5mmol / L; The second washing liquid contains the ethanol of 65v / v%, 100mmol / L 1 L of Tris-HCl and 1 mmol / L of EDTA; the eluent contains 10 mmol / L of Tris-HCl and 10 mmol / L of EDTA.

[0036] The DNA extraction steps are as fol...

example 2

[0044] Using the results of cervical cell sampling in another maternal and child health care center, the cervical cells collected by the cervical brush were placed in 0.8ml of cell preservation solution and mixed evenly to obtain a cell sample solution, wherein the cell preservation solution component was: 3mol / L guanidine hydrochloride , Tris-HCl of 50mmol / L, EDTA of 1mmol / L and Trixon-100 of 7.5v / v%, and the rest is deionized water. After the sample was collected and transported at room temperature for 4 days, a sample was randomly selected for extraction. The components of the extraction reagent were as follows:

[0045] Wherein the first washing liquid contains the ethanol of 60v / v%, the guanidine hydrochloride of 0.1mol / L, the Tris-HCl of 10mmol / L and the EDTA of 1mmol / L; The second washing liquid contains the ethanol of 80v / v%, 10mmol / L Tris-HCl and 10mmol / L EDTA; the eluate contains 100mmol / L Tris-HCl and 1mmol / L EDTA.

[0046] The DNA extraction steps are as follows: ...

example 3

[0054] Using the cervical cell sampling results of another municipal hospital, the cervical cells collected by the cervical brush were placed in 1.2ml of cell preservation solution and mixed evenly to obtain a cell sample solution, wherein the cell preservation solution component was: 2.5mol / L thiocyanate Guanidine acid, Tris-HCl of 25mmol / L, EDTA of 5mmol / L and Trixon-100 of 6v / v%, and the rest is deionized water. After the sample was collected and transported at room temperature for 5 days, a sample was randomly selected for extraction. The components of the extraction reagent were as follows:

[0055] Wherein the first washing liquid contains the EDTA of the ethanol of 55v / v%, the guanidine thiocyanate of 0.6mol / L, the Tris-HCl of 60mmol / L and 9mmol / L; The second washing liquid contains the ethanol of 75v / v%, 80mmol / L Tris-HCl and 8mmol / L EDTA; the eluent contains 50mmol / L Tris-HCl and 5mmol / L EDTA.

[0056] The DNA extraction steps are as follows:

[0057] 1) Take 300 u...

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Abstract

The invention provides a cervical cell preservation and DNA fast extraction integrated kit which comprises cell preservation liquid, a first cleaning solution, a second cleaning solution, eluent and a DNA purification column. The cell preservation liquid comprises 2-3mol / L of guanidine hydrochloride or guanidinium thiocyanate, 5-50mmol / L of Tris-HCl, 1-10mmol / L of EDTA and 5-7.5v / v% of Trixon-100. The cervical cell preservation and DNA fast extraction integrated kit has the advantages that cervical sample DNA can be preserved in the cell preservation liquid of the kit for more than a week under room temperature, transportation of collected samples is facilitated, subsequent DNA extract does not need complex splitting anymore, the whole extraction process can be completed in 10 minutes, and DNA extraction efficiency is increased evidently; in addition, the extraction operation only needs a liquid-moving device and a centrifuge, instrument dependence is low, and the extracted DNA is high in purity.

Description

technical field [0001] The present invention relates to the field of molecular biology. More specifically, the present invention relates to a method for rapidly extracting DNA from cervical cells and virus microorganisms and an integrated kit for preservation and extraction. Background technique [0002] Cervical cancer is the second most common malignant tumor in Chinese women. Studies have shown that more than 99% of cervical cancers are related to human papillomavirus (HPV). At the same time, cervical cancer is the only cancer with a clear cause and early detection and early prevention. Therefore, Screening for HPV plays an important role in preventing cervical cancer. The detection process of HPV involves the sampling, preservation, transportation and DNA extraction of cervical samples. At the same time, since HPV detection is mostly used for clinical detection, simple experimental operations and less dependence on equipment are a must-have feature of the DNA extractio...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10A01N1/02
Inventor 郑岷雪马勇赵国栋刘派李甲
Owner GWP BIOTECHNOLOGIES INC
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