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Method for HCV RNA real-time fluorescent detection with two probes

A technology for hepatitis C virus and ribonucleic acid, which is applied in the field of detecting viral ribonucleic acid, can solve the problems of low sensitivity and unsatisfactory specificity, and achieve the effect of improving sensitivity

Inactive Publication Date: 2007-02-07
AFFILIATED HOSPITAL OF NANTONG UNIV
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AI Technical Summary

Problems solved by technology

[0002] In the prior art, the detection of hepatitis C virus ribonucleic acid (HCVRNA) has Pidge method, Huamei method, etc., all of which have defects such as low sensitivity and unsatisfactory specificity.

Method used

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Embodiment Construction

[0024] A method for real-time fluorescent detection of hepatitis C virus ribonucleic acid (HCVRNA) by two probes, comprising the following steps:

[0025] (1) Preparation of primers and probes: According to the full series of HCV (hepatitis C virus) (GenBank registration number is AB114136), in the 5' non-coding region (5'-UTR) of the HCV (hepatitis C virus) genome, apply Beacon Designer2.1 software, design primer HCV-S, HCV-AS and probe HCV-P1, HCV-P2 of following sequence, and synthesis:

[0026] Primer HCV-S:

[0027] 5'-AACTACTGTCTTCACGCAGAAAGC-3'(nt52~nt75)

[0028] Primer HCV-AS: 5'-CACTCACAAGCGCCCTATCAG-3'(nt312~nt292)

[0029] Probe HCV-P1:

[0030] 5'-ACAGCCTCCAGGCCCCCCC-Fluorescein-3'(nt106~nt124)

[0031] Probe HCV-P1:

[0032] 5′-LC Red 640-CCCGGGAGAGCCATAGTGGTCTG-3′(nt127~nt149)

[0033] (2) Prepare the reaction solution: each part of the reaction solution is 18 μl, and each part of the reaction solution contains the following components:

[0034] 1×PCR Buf...

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PUM

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Abstract

The disclosed dual-probe real-time detection method for hepatitis-C virus RNA comprises: preparing the primer and probe, preparing the reaction solution and RNA sample, and quantitative detecting. This invention improves the sensitivity and specificity for HCVRNA detection.

Description

Technical field: [0001] The invention relates to a method for detecting viral ribonucleic acid. Background technique: [0002] In the prior art, the detection of hepatitis C virus ribonucleic acid (HCVRNA) includes the Pikey method, Huamei method, etc., all of which have defects such as low sensitivity and unsatisfactory specificity. Invention content: [0003] The purpose of the present invention is to provide a method for real-time fluorescence detection of hepatitis C virus ribonucleic acid with two probes that can effectively improve detection sensitivity and specificity. [0004] Technical solution of the present invention is: [0005] A method for two-probe real-time fluorescent detection of hepatitis C virus ribonucleic acid (HCVRNA), is characterized in that: comprise the following steps: [0006] (1) Preparation of primers and probes: According to the full series of HCV, in the 5' non-coding region (5'-UTR) of the HCV (hepatitis C virus) genome, design and synth...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64C12Q1/68
Inventor 崔之础张冬雷施健黄竺筠
Owner AFFILIATED HOSPITAL OF NANTONG UNIV
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