Detection method for high risk type human papilloma virus and reagent case

A technology of human papillomavirus and detection method, which is applied in the field of high-risk human papillomavirus detection method and kit, can solve the problems of low resolution, weak target signal, strong background interference, etc., to reduce the interference of background signal, The effect of increasing the number of molecules and improving sensitivity

Inactive Publication Date: 2007-12-05
海南森瑞谱生命科学药业股份有限公司
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  • Claims
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AI Technical Summary

Problems solved by technology

[0010] The technical problem to be solved by the present invention is that, for the low detection specificity, false positive results and low-density DNA chip technology instability of the above-mentioned existing PCR technology and its direct primer hybridization, the target signal is weak, the background interference is strong, and the resolution is low To provide a new high-risk human papillomavirus detection method and kit

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0063] (1) Preparation of chips

[0064] Select the glass slide as the chip carrier, dilute the high-risk HPV virus probe with deionized water, and dissolve it in 500mmol / L Na 2 CO 3 / NaHCO 3 Prepare a sample with a final concentration of 10 nmol / L in the solution, add the sample to the spotting plate, fix it at room temperature for 48-72 hours, and dry it for later use. The chip monitoring system consists of a blank spot, a positive internal reference and a negative internal reference. The average value of the positive fluorescent signal exceeds three times of the signal of the negative point to be judged as a positive standard.

[0065] (2) DNA preparation

[0066] DNA extraction kits were used and stored at 4°C.

[0067] (3) Amplification of samples to be tested

[0068] 1. Take a 0.2ml EP tube, add 16.5ul of sterile double distilled water, 2.5ul of 10 times PCR reaction buffer, 0.5ul of dNTP, 1ul each of specific primers NP1, NP2, NP3 and NP4 under ice bath condition...

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PUM

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Abstract

The present invention is high risk human papilomavirus (HPV) detecting process, which includes the following steps: 1. designing and synthesizing proper specific primer, and amplifying the target DNA of the detected sample specifically for the second time by means of one-step nest PCR technology; and 2. hybridizing and selectively typing the target DNA of the detected sample through twice specific selective amplification and the specific segment of high risk HPV by means of combination to low density DNA chip technology, and signal identification through fluorescent detecting or direct color developing. The present invention provides also one corresponding kit. The present invention can amplify specific target DNA segments of 13 kinds of HPV simultaneously and detect several kinds of virus subtypes of HPV through once reaction, and has the advantages of high speed, high efficiency, high sensitivity, high specificity, etc.

Description

technical field [0001] The invention relates to the field of molecular biology, in particular to a detection method and kit for high-risk human papillomavirus. Background technique [0002] Cervical cancer is a malignant tumor that seriously threatens women's lives. Once it occurs, its survival and cure rates are very low. Scientific research has confirmed that human papillomavirus (HPV) is a necessary condition for causing cervical cancer, and 99.7% of cervical cancer cases have been found to be infected with high-risk HPV viruses. Because the high-risk HPV virus that causes cervical cancer is a weak virus, if the infection can be detected in time and treated in time, the virus can be controlled or eliminated, and the purpose of preventing and treating cervical cancer can be achieved. Therefore, to control the occurrence of cervical cancer, the focus is on prevention, and the key is early screening. my country has a large population and uneven economic development. Especi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68G01N21/64
Inventor 张春发邓柳红
Owner 海南森瑞谱生命科学药业股份有限公司
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