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Primer probe group and kit for jointly detecting hepatitis B virus and hepatitis C virus based on fluorescent RMA method

A technology of hepatitis B virus and hepatitis C virus, which is applied in the field of biotechnology detection, can solve the problems of expensive equipment, unsuitability for clinical testing and large-scale blood screening, and complicated operation, so as to reduce the cost of testing, The effect of avoiding false positive results and improving detection efficiency

Inactive Publication Date: 2021-05-14
济南国益生物科技有限公司
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Problems solved by technology

[0005] Although the methods for synchronous detection of HBV and HCV have been reported both at home and abroad, most of them use real-time fluorescent quantitative PCR and gene chip technology, etc., the required equipment is expensive, the operation is complicated, and professional technicians are required to operate, so it is not suitable for use. For clinical testing and large-scale blood screening

Method used

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  • Primer probe group and kit for jointly detecting hepatitis B virus and hepatitis C virus based on fluorescent RMA method
  • Primer probe group and kit for jointly detecting hepatitis B virus and hepatitis C virus based on fluorescent RMA method
  • Primer probe group and kit for jointly detecting hepatitis B virus and hepatitis C virus based on fluorescent RMA method

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Embodiment 1

[0044]1, preparation of positive standard plasmid

[0045]The nucleic acid of HBV and HCV was extracted as a template, and the specific gene of HBV and HCV was used for PCR amplification. The PCR amplification product was electrophoresed by 1% agarose gel electrophoresis, cloning recovery, clone to the PMD18-T vector, transformed to the large intestine Bacillobacteriosis cells, blue-white spot screening, picking white colonies, colonies PCR verification. The positive recombinant bacteria sequencing, and cultivating the correct recombinant bacteria in sequencing overnight, extracting the plasmid DNA, and obtaining the yangotoma.

[0046]2, fluorescent RMA primers and probes design

[0047]According to the HBV sequence conserved surface antigen S gene, HCV sequence conserved NS3 gene designed specific fluorescent RMA primers and probes, as shown in Table 1:

[0048]Table 1 primer and probe sequence

[0049]

[0050]Note: The fluorescent group of the HBV detection probe was modified with VIC modificatio...

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Abstract

The invention relates to the field of biotechnology detection, in particular to a primer probe group and a kit for jointly detecting hepatitis B virus and hepatitis C virus based on a fluorescent RMA method. The primer probe group comprises a primer and a probe for detecting hepatitis B virus, and a primer and a probe for detecting hepatitis C virus; a fluorescence reporter group marked on the HBV detection probe is VIC, and a fluorescence quenching group is BHQ1; a fluorescence reporter group marked on the HCV detection probe is CY5, and a fluorescence quenching group is BHQ2; the kit comprises a detection tube containing an amplification reaction reagent, a buffer solution, magnesium acetate, a standard positive plasmid and sterile double distilled water.

Description

Technical field[0001]The present application is a primary detection of biotechnology detection, specifically a primer probe group and kit for detecting hepatitis B virus and hepatitis C virus based on fluorescence RMA method.Background technique[0002]Hepatitis B virus (HBV) is a DNA virus belonging to the liver DNA virus. my country is a high-risk area of ​​HBV infection. Although since 2006, the vaccination plan of hepatitis B has declined year by year, but compared with developed countries (HBV infection rate <2%), my country's HBV infection rate is still Higher. HBV infections can cause acute and chronic liver diseases, including cirrhosis and liver cancer.[0003]Hepatitis C virus (HCV) is a single stranded RNA virus belonging to the yellow strain department. Hepatitis C is caused by HCV infection, mainly spread by blood and body fluid, accounting for 70% of hepatitis after blood transfusion. In my country, HCV is secondary only to HBV factors caused by chronic hepatitis. Long-...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/6844C12N15/11C12R1/93
CPCC12Q1/6844C12Q1/706C12Q1/707C12Q2600/16C12Q2521/507C12Q2522/101C12Q2531/119C12Q2563/107C12Q2537/143C12Q2547/101
Inventor 陈大为陈龙张瑶
Owner 济南国益生物科技有限公司
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