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Gene chip for inspecting HIV P-reverse transcriptase inhibiting resistance and its reagent kit

A technology of reverse transcriptase inhibition and gene chip, applied in the field of detection, can solve the problems of heavy workload, time-consuming, expensive, etc., and achieve the effect of strong repeatability, easy operation and high accuracy

Active Publication Date: 2010-05-05
TIANJIN BIOCHIP TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The cost of genotype detection is relatively low, and the technology is relatively easy. The results can be obtained within 1 to 2 weeks after sample collection. Phenotype detection can directly detect the drug resistance of the virus, but the operation is complicated, time-consuming, technically demanding, and very difficult. Expensive, so genotypic drug resistance detection is currently widely used clinically in the world
Among genotypic drug resistance detection methods, DNA sequence analysis is the most accurate method, but it is labor-intensive, expensive and time-consuming

Method used

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  • Gene chip for inspecting HIV P-reverse transcriptase inhibiting resistance and its reagent kit
  • Gene chip for inspecting HIV P-reverse transcriptase inhibiting resistance and its reagent kit
  • Gene chip for inspecting HIV P-reverse transcriptase inhibiting resistance and its reagent kit

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Embodiment one Preparation of oligonucleotide probes

[0038] 1. Probe design: Based on 163 strains (3 of which are HXB2, U71182 and AY531116, which are representative strains of HIV in China; the other 160 sequences were provided by the National AIDS Center of the Chinese Center for Disease Control and Prevention, which were isolated in China in 2004. The comparison result of the bioinformatics of the HIV drug-resistant strain) HIV RT gene, utilize Primer Premier 5.0 bioinformatics software to design and screen out 96 probes (as shown in Table 1) for 14 sites, screen It is mainly based on the following two points: one is to place the detection site at the center of the probe, and the other is to keep the Tm values ​​of all probes consistent.

[0039] Oligonucleotide probes screened out in table 1

[0040]

[0041]

[0042]

[0043] 2. Probe synthesis: Synthesize the probe sequence designed in Table 1 and modify the 5' terminal amino group and poly(T).

Embodiment 2

[0044] Embodiment two Preparation of primer sequences for amplifying HIV RT gene

[0045] Using Premier 5.0 bioinformatics software, the PCR amplification primers shown in Table 2 were designed and synthesized using HXB2 as a template.

[0046] Table 2 is used for the primer sequence of amplification HIV RT gene

[0047] SEQ ID

[0048] SEQ ID

Embodiment 3

[0049] Embodiment three Preparation of gene chip

[0050] The probes were placed on the biochip spotter (Spotarray 72) figure 1 The arrangement shown (A, B, C, and D four array probes are arranged in the same way) was spotted on the aldehyde-formed glass slide with a specification of 25mm×75mm×1mm (width×length×height). Thus making a gene chip.

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Abstract

Gene chip for inspecting HIV to reverse transcriptase inhibitor resistance and reagent kit are disclosed. The gene chip consists of solid-phase carrier and oligonucleotide probe fixed on the solid-phase carrier, which comprises mutation site DNA fragment selected in HIV reversed transcriptive enzyme gene area, PCR amplifying for sample cDNA to be inspected by primer, purification marking for amplified product and hybridizing with the gene chip, acquiring hybridized signal by gene chip scanner, analyzing and appraising to determine medicine-tolerance site mutation. It's simple, accurate and repeated.

Description

technical field [0001] The invention belongs to the technical field of detection, and relates to a gene chip (or DNA microarray) and a method for using the same and a detection kit, in particular to a human immunodeficiency virus (humanimmunodeficiency virus Type 1 HIV-1) reverse transcriptase (reverse transcriptase, RT) A gene chip for detection of point mutations in the gene in China for HIV resistance to reverse transcriptase inhibitors, a detection method and a detection kit. Background technique [0002] AIDS, or Acquired Immunodeficiency Syndrome (AIDS), is a group of devastating infections caused by the human immunodeficiency virus, HIV, which attacks and destroys certain white blood cells important to the body's immune system. In 1985, China reported its first AIDS patient. At present, China's AIDS is in a situation of low prevalence in the whole country and high prevalence in some areas and specific groups. As of the end of September 2004, there were 89,067 report...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68
Inventor 王磊刘丹许天正王金忠
Owner TIANJIN BIOCHIP TECH CO LTD
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