Human papilomavirus typing gene chip detecting system

A technology of human papillomavirus and gene chip, which is applied in the field of gene chip; can solve the problem that the limitation of detection flux cannot meet the clinical needs and other problems

Active Publication Date: 2010-05-05
亚能生物技术(深圳)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the common PCR method and fluorescent quantitative PCR can...

Method used

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  • Human papilomavirus typing gene chip detecting system
  • Human papilomavirus typing gene chip detecting system
  • Human papilomavirus typing gene chip detecting system

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0095] [Example 1] Preparation of HPV Typing Detection Chip

[0096] 1. Manufacturers and specifications of raw materials

[0097] name

manufacturers

Specification

Product name

EDAC

Sigma

Nylon membrane

Pall

Pore ​​size: 0.45uM

30cm×3M

Biodyne C

Membranes

[0098] The probe sequence design is shown in SEQ ID 1-36, and the 3'-terminal amino-labeled probe was synthesized by Shanghai Boya Biotechnology Co., Ltd. according to methods well known to those skilled in the art.

[0099] 2. Production of membrane strips

[0100]

Embodiment 2

[0101] [Example 2] Sample detection of HPV typing chip detection kit

[0102] From August to October 2003, clinical verification was carried out in Shenzhen Hospital of Peking University and Shenzhen People's Hospital respectively. The results proved that the detection system of the present invention has large throughput and high sensitivity. Quantitative PCR comparable sensitivity. The specific reagents and detection methods are as follows:

[0103] ●Kit composition

[0104]

[0105] Wherein the PCR Mix contains primers of SEQ ID NO.37-38.

[0106] ●Necessary equipment and prepared reagents

[0107] PCR instrument DNA electrophoresis instrument

[0108] Molecular Hybridization Box Shaker

[0109] 3%H 2 o 2

[0110] Other required reagents were prepared according to the following formula:

[0111] 20×SSC (the pH value of the solution directly affects

sound color results)

NaCl 175.3g

Sodium citrate 88.2g

Add H 2 O dissolved to 1000mL, ...

Embodiment 3

[0144] [Example 3] Comparison of the hybridization effect of the 3'-terminal amino-labeled probe and the 5'-terminal amino-labeled probe

[0145] The hybridization probe can be amino-labeled at the 3’ or 5’ end of the oligonucleotide. The following experiments compare the hybridization effects of the 3’ and 5’ amino-labeled probes:

[0146] Synthesize 3'-terminal amino-labeled probes and 5'-terminal amino-labeled probes respectively, prepare nylon membranes with the method in Example 1, and carry out hybridization according to the method of Example 2 with positive control samples and negative control samples, the results are shown in image 3 ,Depend on image 3 The results show that under the same conditions, the labeling effect of the 3'-terminal amino group is better than that of the 5'-terminal labeling effect; specificity.

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PUM

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Abstract

The invention relates to the gene-chip detection system for the HPV parting, including the gene chip which is used to detect the separate hypotype of HPV, and the increase of PCR primer of the separate hypotype nucleic acid of HPV. The gene chip contains a substratum and a scanning-probe fastened on the substratum, and the scanning-probe is nucleotide which can be hybrid with the nucleic acid of the separate hypotype virus of HPV. The invention constructs a gene-chip detection system for HPV parting contraposing 15 kinds of high risk hypotype and 3 kinds of low risk hypotype of HPV, with whichthe HPV hypotype in the clinical sample can be detected speedily and precisely. It has an important significance to the clinical diagnosis of HPV and the early prevention against uterine neck cancer,and can be extensively used to high effectively diagnose and to the female body-check.

Description

technical field [0001] The invention relates to a gene chip containing probes of various subtypes of human papillomavirus (HPV); [0002] The invention also relates to detection reagents for detecting various subtypes of human papillomavirus in samples. Background technique [0003] Papillomaviruses (Papillomaviruses) are epitheliophilic viruses, widely distributed in humans and animals, highly specific, and there is no cross-infection between species. Human papillomaviruses (Human Papillomaviruses, HPV), a DNA virus, are a class of papillomaviruses that infect the stratified squamous epithelium of human skin and mucous membranes. There are currently more than 100 types of HPV identified. According to the risk of different types of HPV and cancer, it is divided into low-risk HPV such as HPV6, 11, 42, 43, 44, etc., which often cause benign lesions such as external genital warts, including low-grade cervical intraepithelial lesions (CIN1), high-risk Types of HPV such as HPV...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 张旭向筑周逸
Owner 亚能生物技术(深圳)有限公司
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