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Gene chip, reagent and kit thereof for high-throughput typing detection of human papilloma virus

A detection kit and gene chip technology, applied in the field of gene chips, can solve the problems of complex operation, large area of ​​membrane strip carrier, and inability to detect quickly.

Active Publication Date: 2013-11-27
GENETEL PHARMA SHENZHEN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the reverse dot hybridization method commonly used in PCR-based detection methods for multiple types of HPV attaches DNA probes for HPV detection to a larger membrane strip (several centimeters to more than ten centimeters) or other substrates. Membrane strips have disadvantages such as large carrier area, large hybridization volume, long hybridization time, the need to use a rotary hybridization instrument, and complicated operations. Parallel high-throughput fast detection

Method used

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  • Gene chip, reagent and kit thereof for high-throughput typing detection of human papilloma virus
  • Gene chip, reagent and kit thereof for high-throughput typing detection of human papilloma virus
  • Gene chip, reagent and kit thereof for high-throughput typing detection of human papilloma virus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Embodiment 1: Preparation of HPV gene chip microarray

[0036] Embodiment 1 has exemplified the preparation method of HPV gene chip microarray, figure 1 is the arrangement matrix of HPV probes and control point probes in each gene chip, the diameter of each point is 150 μm, and the center-to-center distance of each point is 500 μm. For the sample printing process of the chip, see figure 2 , the Pall Biodyne C membrane is cut into the size of each film strip 24cm × 20cm, after pretreatment with EDC, prepare SEQ ID NO.1 to SEQ ID listed in the sequence list with 0.1M phosphate buffer (pH7.4) No.30 probe sample solution, the probe concentration is 0.2μM-5μM, use the Heath Digit2003 biochip spotting instrument to place the probe according to figure 1 The medium HPV chip microarray array is arranged in a matrix, and the spray point is in the chip with an area of ​​6.4mm×6.4mm. image 3 For example, a part of the microarray scanning image (5×5 matrix) after spraying is sho...

Embodiment 2

[0037] Example 2: Sample detection experiment using human papillomavirus typing detection kit (gene chip method) (hereinafter referred to as "HPG" reagent)

[0038] 【Product Name】Common Name: Human Papillomavirus Typing Detection Kit (Gene Chip Method)

[0039] [packing specification] 48 tests / box

[0040] 【Principle of Inspection】

[0041] This kit adopts gene chip detection technology. Use micro-spotting technology to spot HPV type-specific probes on the gene chip substrate to make a gene chip; after PCR amplification, hybridization and color development of the DNA in the sample, it is detected and read by the HPV typing gene chip The system automatically collects images and analyzes and reports the detection results. The main components and detection process of the kit are as follows:

[0042] 【Main components】

[0043] This kit consists of A box and B box (see Table 1); A box includes 5 kinds of PCR-related components, and B box includes 9 kinds of hybridization and c...

Embodiment 4

[0158] Embodiment 4 compares with the internationally general HPV detection method

[0159] 1. Results compared with Digene HC2

[0160]Digene HC2 is the second-generation hybridization capture method of Digene Corporation in the United States, including two sets of probes, which can detect 13 high-risk types (16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59 , 68) and 5 low-risk types (6, 11, 42, 43, 44), which are the earliest HPV DNA detection products approved by the FDA in the United States, and are recognized as the "gold standard" by the industry. Domestically introduced mainly detect 13 high-risk types Type HC2 reagent, for the sake of simplicity, the gene chip detection kit of the present invention is referred to as "HPG" for short, the following is the comparative detection results of HPG and HC2 (high risk): ( Description: Samples in the table below The serial number is recoded for the source, not the original serial number ). The genotyping comparison results of ...

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Abstract

The invention discloses a gene chip, a reagent and a kit for high-throughput typing detection of a human papilloma virus. The gene chip comprises a carrier membrane substrate, and a probe fixed on the substrate, wherein the probe is nucleotide which can hybridize with various types of DNA special segments of human papilloma virus; 5' end of the probe is labeled by amino, and a nucleotide sequence of the probe is any one of probe nucleotide sequences in an attached sequence list. The invention provides the gene chip for detecting various types of the human papilloma virus, and simultaneously provides a detection reagent for rapid and parallel detection of various types of HPVs (human papilloma virus) as well as for high-throughput analysis of a great sample size.

Description

technical field [0001] The technical field of the present invention relates to a gene chip technology; in particular, it relates to a gene chip for detecting human papillomavirus (HPV) DNA and its reagents, and also relates to a reagent for detecting various types of HPV DNA in a sample by using the method box. Background technique [0002] Human papillomavirus (HPV) is a type of DNA virus that infects human epithelial cells. According to the definition of the nucleotide sequence homology of the open reading frames in the E6, E7 and L1 regions of the HPV genome, more than 100 HPV types have been discovered. , some types can be further divided into different subtypes according to the definition of the above-mentioned regional homology of 90%-98% and not exceeding 98%. [0003] HPV can cause skin or mucous membrane infections in various parts of the human body, leading to a variety of epithelial tissue diseases, such as skin common warts, flat warts, and genital warts. Some ...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68C40B40/06
Inventor 黄明辉黄鹤李勐赵雷
Owner GENETEL PHARMA SHENZHEN
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