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HPV high risk and low risk subtype typing DNA micro-array chip

A microarray chip, a technology for risk subtyping, which is applied in the determination/examination of microorganisms, biochemical equipment and methods, etc. performance, flux issues, etc.

Active Publication Date: 2009-07-22
DAAN GENE CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These traditional HPV detection methods have poor sensitivity and specificity, high false positive and false negative rates, and cannot be typed
[0008] In recent years, with the development of molecular biology, scientists have adopted modern molecular biology methods, including computational hybridization, PCR, and fluorescent quantitative PCR, for HPV detection, but these methods still have sensitivity, specificity, and throughput. and other aspects of the problem, can not be well adapted to clinical requirements

Method used

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  • HPV high risk and low risk subtype typing DNA micro-array chip
  • HPV high risk and low risk subtype typing DNA micro-array chip
  • HPV high risk and low risk subtype typing DNA micro-array chip

Examples

Experimental program
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Effect test

Embodiment 1

[0030] Method 1: Design of PCR primers and probes for HPV high-risk and low-risk subtype typing DNA microarray chips

[0031] In the L region where the polymorphisms of the HPV genome sequence are concentrated, design and synthesize specific PCR primers, 5' end primers PH1, PH1-1, and CY3 markers; 3' end primers P2-1, P2-2, P2-3, P2-4, P2-5, P2-6, the primer sequences are shown in Table 2. Multiplex asymmetric PCR was used to amplify in one tube.

[0032] According to the polymorphic characteristics of each subtype gene sequence in the HPV L region, 28 pairs of subtype probes were designed and synthesized, and a pair of positive and negative control probes were also designed. See Table 1 for subtyping probe sequences.

[0033] Method 2: Preparation of HPV high-risk and low-risk subtype typing DNA microarray chips

[0034] Microarray chip preparation requires one of each pair of probes. Using a gene chip automatic spotting instrument, 28 typing probes and one of the two con...

Embodiment 2

[0037] Embodiment 2: Use the product of the present invention to detect 10 clinical samples

[0038] Take 10 samples, extract virus DNA, and perform DNA sequencing first. The sequencing results are as follows:

[0039] sample number Sequencing results (HPV type) sample number Sequencing results (HPV type) 01 16 06 feminine 02 18 07 feminine 03 11 08 feminine

[0040] 04 16，18 09 feminine 05 11，16 10 feminine

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Abstract

The invention relates to DNA microarray chips, in particular to DNA microarray chips under human papilloma virus (HPV) high-risk and low-risk subtype classification. A solid-phase carrier substrate is adopted and matched with oligonucleotide probes of each HPV subtype so as to prepare the DNA microarray chips, which can simultaneously detect the HPV high-risk and low-risk subtypes of multiple samples at one time in high sensitivity and high specificity, and can be widely applied into qualitative and subtype detection of HPV infection.

Description

Technical field [0001] The invention relates to a DNA microarray chip, in particular to a human papillomavirus (HPV) subtype genotyping DNA microarray chip. Using a solid-phase carrier substrate, the oligonucleotide probes for each subtype of HPV are prepared into a DNA microarray chip, which can simultaneously detect high-risk and low-risk subtypes of HPV in multiple samples at one time with high sensitivity and high specificity , can be widely used in HPV virus infection qualitative and typing detection. technical background [0002] Human papilloma virus (HPV) is a minimal DNA (deoxyribonucleic acid) virus. Mainly infects epithelial cells. So far, more than 80 subtypes of HPV have been identified, and more than 100 subtypes of HPV have been reported. HPV has a high infection rate in humans, from less than 1% to as high as 50% in natural populations, and more than 20% to 80% of sexually active people have a history of HPV infection. [0003] One of the most common part...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/70
Inventor 胡守旺张帆李明程钢何蕴韶
Owner DAAN GENE CO LTD
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