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Hepatitis C virus (HCV) genotyping kit

A hepatitis C virus, genotyping technology, applied in the direction of microbe-based methods, microbiological measurement/testing, microbiology, etc., can solve the problems of low specificity, high equipment requirements, long detection time, etc., and achieve high sensitivity , low false negative rate, and high accuracy

Active Publication Date: 2014-08-06
福州泰普生物科学有限公司
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Problems solved by technology

[0004] At present, the genotyping methods of HCV mainly include: restriction length polymorphism analysis (RFLP), genotype-specific primer PCR method, genotype-specific probe nucleic acid hybridization analysis method, direct sequencing method and other methods, but these methods Most of them have the disadvantages of cumbersome operation, high requirements for equipment, long detection time, low throughput, low sensitivity and low specificity, etc.

Method used

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  • Hepatitis C virus (HCV) genotyping kit
  • Hepatitis C virus (HCV) genotyping kit
  • Hepatitis C virus (HCV) genotyping kit

Examples

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Embodiment

[0065] 1. Design RT and PCR primers in the 5'UTR region of the HCV genome, and use an asymmetric RT-PCR system to amplify the target fragments required for typing; as shown in Table 1, design a type 1 probe within the range of the target fragments. 3 needles, 4 type 2 probes, 3 type 3 probes, 3 type 6 probes and 1 universal probe.

[0066] 2. Please refer to figure 1 The nylon membrane is printed in a grid according to the designed format, and 14 typing probes and 2 control probes are spotted on the specific area of ​​the membrane strip by using an automatic film spotter to make a DNA hybridization membrane strip.

[0067] Spotting probe solution: dilute the probe into 5×SSC, 0.05% SDS solution, the final concentration is 10 μM;

[0068] Spotting matrix: Spot the probes on the membrane strip, each probe spotting 0.5pM, 8 points per row, 2 rows in total (see figure 1 ).

[0069] 3. Use the product of the present invention to detect clinical samples

[0070] Take 8 cases of ...

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Abstract

The invention relates to a disease pathogen gene detecting technology and particularly relates to a hepatitis C virus (HCV) genotyping kit which can be used for auxiliary diagnosis for infection of types 1a, 1b, 2a, 2b, 3a, 3b and 6a of HCV happening in China. According to the invention, a nylon membrane carrier is adopted, an oligonucleotide probe is designed according to the different genotypes of the HCV, a gene chip is prepared, and then, a PCR (Polymerase Chain Reaction) primer and other components are matched, so that the HCV in a blood sample can be rapidly and accurately typed. The HCV genotyping kit provided by the invention has the characteristics of rapidness, sensitivity, accuracy, high flux, simplicity, convenience, high sensitivity and strong specificity, and is capable of detecting various target sequences in hybridization reaction once, convenient to take materials, free of special equipment and low in instrument investment.

Description

technical field [0001] The present invention relates to a gene detection technology of disease pathogens, in particular to a hepatitis C virus genotyping detection kit, which is used to detect 1a, 1b, 2a, 2b, 3a, 3b and 3b of hepatitis C virus occurring in China. Auxiliary diagnosis of type 6a infection. Background technique [0002] Hepatitis C is distributed worldwide and the population is generally susceptible to it. It is an infectious disease that seriously endangers human health and spreads widely. Its pathogen is hepatitis C virus (HCV). Currently, its infection rate ranges from 0.1% to 10%, with an average of 3%. More than 170 million people are infected with HCV in the world, and more than 100,000 HCV patients develop liver cancer every year, followed by gastrointestinal bleeding and ascites. According to the WHO 2002 annual report, in 2001, chronic liver disease caused 14 million deaths, including 796,000 cases caused by liver cirrhosis and 616,000 cases caused b...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68C12R1/93
CPCC12Q1/6834C12Q1/70C12Q2531/107C12Q2545/101
Inventor 彭春梅谢佐福姚铭锋
Owner 福州泰普生物科学有限公司
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