Fluorescent quantitative DCR kit for detecting swine pseudo rabies virus and its use

A porcine pseudorabies virus and fluorescence quantitative technology, which can be used in the determination/inspection of microorganisms, biochemical equipment and methods, etc., can solve the problems of long cycle, cumbersome operation, and difficult detection of pseudorabies.

Active Publication Date: 2005-12-21
常州同泰生物药业科技股份有限公司
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  • Abstract
  • Description
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  • Application Information

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Problems solved by technology

[0003] Traditional pseudorabies detection methods mainly include: enzyme-linked immunosorbent assay (ELISA) test, agar immunodiffusion test, virus hemagglutination (HA) and hemagglutination inhibition test (HI), colloidal gold and other methods. Import and export inspection and quarantine play an important role, but there are common obvious deficiencies, such as long cycle, cumbersome operation, poor specificity and sensitivity, non-standard, not suitable for large-scale detection, which brings great difficulties to the detection of pseudorabies

Method used

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  • Fluorescent quantitative DCR kit for detecting swine pseudo rabies virus and its use
  • Fluorescent quantitative DCR kit for detecting swine pseudo rabies virus and its use
  • Fluorescent quantitative DCR kit for detecting swine pseudo rabies virus and its use

Examples

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Effect test

Embodiment 1

[0028] Example 1 Detecting pseudorabies virus with a fluorescent quantitative PCR kit for rapid quantitative detection of pseudorabies virus

[0029] 1 Composition and preparation of porcine pseudorabies virus fluorescent quantitative PCR kit,

[0030] Reagent composition:

[0031] a). DNA extraction reagent is prepared by yourself, lysate: 4mol / L guanidine isothiocyanate, 0.5% sodium lauryl sarcosinate, 0.1mmol / L β-mercaptoethanol, 25mmol / L sodium citrate, isopropanol , double distilled water, 75% ethanol prepared with sterile double distilled water.

[0032] b). Standard positive template.

[0033] c). Fluorescence quantitative reaction solution: PCR 10×buffer 2μl, forward primer and reverse primer 1.5μl (10μmol / L), fluorescent probe 2μl (1μmol / L), MgCl 2 2.5μl (25mmol / L), dNTPs 0.5μl (10mmol / L), Taq DNA polymerase 0.5μl (3U / μl), sterile double distilled water 9μl. The FP sequence of the fluorescent probe is SEQ ID NO: 3, the fluorescent reporter group labeled at the 5' ...

Embodiment 2

[0042] Example 2 Detection of pseudorabies virus with a fluorescent quantitative PCR kit for rapid quantitative detection of pseudorabies virus

[0043] Clinical specimens: From February 2005 to May 2005, nasal swabs and tissue samples were collected from pigs in Beijing, Xiantao, Hubei, and Minggang, Henan. age).

[0044] 1 Composition and preparation of porcine pseudorabies virus fluorescent quantitative PCR kit,

[0045] Reagent composition:

[0046] a). DNA extraction reagents are prepared by yourself, lysate: 4mol / L guanidine isothiocyanate, 0.5% sodium lauryl sarcosinate, 0.1mmol / L β-mercaptoethanol, 25mmol / L sodium citrate, isopropyl Alcohol, double distilled water, 75% ethanol prepared with sterile double distilled water.

[0047] b). Standard positive template.

[0048] c). Fluorescence quantitative reaction solution: PCR 10×buffer 2μl, forward primer and reverse primer 1.5μl (10μmol / L), fluorescent probe 2μl (1μmol / L), MgCl 2 2.5μl (25mmol / L), dNTPs 0.5μl (10mmo...

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Abstract

This invention discloses a kind of measuring PRVHS fluorescence quantitative PCR reagent box and application. The reagent box has a) DNA cracking liquid, b)Taq DNA polymerization enzyme, c) pG-HB-contained nucleoside series of standard positive template, d)the fluorescence quantitative reaction liquid containing positive and negative primer and fluorescence probe series. Advantages: simple and easy for operation, fast and accurate quantitation and so on.

Description

technical field [0001] The invention relates to a fluorescent quantitative PCR kit for quantitative detection of porcine pseudorabies virus, which is suitable for qualitative and quantitative detection of porcine pseudorabies virus, and also relates to applications for qualitative and quantitative detection of porcine pseudorabies virus. Background technique [0002] Pseudorabies is a highly contagious infectious disease of pigs, which often causes serious economic losses to the pig industry. At present, the early specific diagnosis of the disease, the culling and vaccination of infected animals are important means to effectively prevent and control swine fever. [0003] Traditional pseudorabies detection methods mainly include: enzyme-linked immunosorbent assay (ELISA) test, agar immunodiffusion test, virus hemagglutination (HA) and hemagglutination inhibition test (HI), colloidal gold and other methods. Import and export inspection and quarantine ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 潘兹书张楚瑜万超
Owner 常州同泰生物药业科技股份有限公司
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