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.
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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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