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Two-color fluorescence quantitative polymerase chain reaction (PCR) combined detection method of swine fever virus and blue ear disease virus and kit thereof

A porcine PRRS virus and dual-color fluorescence technology are applied in the detection field of swine fever virus and porcine PRRS virus, which can solve problems such as incomplete joint detection methods, avoid easy missed diagnosis and misdiagnosis, enhance sensitivity and specificity, Apply convenient effects

Active Publication Date: 2011-10-12
广东省农业科学院兽医研究所
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  • Abstract
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  • Application Information

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Problems solved by technology

At present, the combined detection method for the infection of CSFV and porcine PRRSV has not been perfected yet

Method used

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  • Two-color fluorescence quantitative polymerase chain reaction (PCR) combined detection method of swine fever virus and blue ear disease virus and kit thereof
  • Two-color fluorescence quantitative polymerase chain reaction (PCR) combined detection method of swine fever virus and blue ear disease virus and kit thereof
  • Two-color fluorescence quantitative polymerase chain reaction (PCR) combined detection method of swine fever virus and blue ear disease virus and kit thereof

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Embodiment 1

[0068] 1. Design of specific primers and probes

[0069] According to the nucleic acid sequences of classical swine fever virus (Classical Swine Fever Virus, CSFV) and porcine blue ear disease virus (Reproductive and Respiratory Syndrome Virus) retrieved from Genbank, the primers and Probe, its sequence is as follows:

[0070] Swine Fever Virus:

[0071] Upstream primer: CSFV-F: 5'-AGCTCCCTGGGTGGTCTAAGT-3' (SEQ ID NO.1);

[0072] Downstream primer: CSFV-R: 5'- CCCTCGTCCACATAGCATCT-3' (SEQ ID NO.2);

[0073] Fluorescent probes:

[0074] CSFV-P: 5'-FAM-AGTTCGACGTGAGCAGAAGCCCACC-TAMRA-3' (SEQ ID NO.3), the amplified fragment is 93 bp;

[0075] Porcine PRRS virus:

[0076] Upstream primer: PRRSV-F: 5'- GGACACCAAGGGCAGACTCT-3' (SEQ ID NO.4);

[0077] Downstream primer: PRRSV-R: 5'-CACCTCCAACCTCAATTTTACC-3' (SEQ ID NO.5);

[0078] Fluorescent probes:

[0079] PRRSV-P: 5'-VIC-CTGGCGGTCACCCGTCATCATAGAG-TAMRA-3' (SEQ ID NO.6), the amplified fragment is 77 bp.

[0080] The fluo...

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Abstract

The invention discloses a two-color fluorescence quantitative polymerase chain reaction (PCR) combined detection method of swine fever virus and blue ear disease virus. By adopting the method, swine fever virus and blue ear disease virus in one sample to be detected can be detected and the loading level of the two viruses can be detected quantitatively. The invention also provides a kit used for the two-color fluorescence quantitative PCR combined detection of swine fever virus and blue ear disease virus. The detection method and kit are convenient and fast to operate and have high specificity and sensitive and reliable detection effect; and the minimum detection concentration is 1*10<2>copy / mu l.

Description

technical field [0001] The present invention relates to a detection method for swine fever virus and porcine blue ear disease virus, in particular to a combined detection method for swine fever virus and porcine blue ear disease virus two-color fluorescent quantitative PCR, and further relates to a detection method for swine fever virus and porcine blue ear disease virus Two-color fluorescent quantitative PCR combined detection kit for virus. Background technique [0002] Classical Swine Fever Virus (CSFV) is a highly contagious febrile infectious disease caused by classical swine fever virus (CSFV). The main manifestations are characteristic pathological changes, visceral hemorrhage, infarction and necrosis, and the mortality rate is very high, causing significant economic losses to the pig industry. Sick pigs are the main source of infection, and direct contact between susceptible pigs and sick pigs is the main way of virus transmission. In my country, the epidemic of sw...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68G01N21/64
Inventor 魏文康温肖会吕殿红黄忠周秀蓉罗胜军贾春玲袁洁温晓慧钱钢锐
Owner 广东省农业科学院兽医研究所
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