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RT-PCR (Reverse Transcription-Polymerase Chain Reaction) primer for detecting European and American porcine reproductive and respiratory syndrome viruses and kit

A technology of porcine PRRS virus and RT-PCR, which is applied in the field of RT-PCR primers and kits for the detection of European and American types of porcine PRRS virus, can solve the problems of genome sequence and antigenicity differences, and avoid false positives. Negative results, high sensitivity, and the effect of saving labor time

Inactive Publication Date: 2014-02-12
广西壮族自治区动物疫病预防控制中心 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although PRRSV can cause the same disease, there are great differences in genome sequence and antigenicity between the European and American strains, and serological experiments show that there is a certain cross-reaction

Method used

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  • RT-PCR (Reverse Transcription-Polymerase Chain Reaction) primer for detecting European and American porcine reproductive and respiratory syndrome viruses and kit
  • RT-PCR (Reverse Transcription-Polymerase Chain Reaction) primer for detecting European and American porcine reproductive and respiratory syndrome viruses and kit
  • RT-PCR (Reverse Transcription-Polymerase Chain Reaction) primer for detecting European and American porcine reproductive and respiratory syndrome viruses and kit

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Embodiment Construction

[0042] The present invention will be further described below in conjunction with embodiment, but do not limit protection scope and application scope of the present invention:

[0043] 1. Kits for detecting European and American types of porcine PRRS virus

[0044] A test kit for detecting European type and American type of porcine PRRS virus, comprising the following reagents, see Table 2 for details, and all reagents used can be purchased from the market.

[0045]Table 2 Detection kits for European type and American type of porcine PRRS virus

[0046]

[0047]

[0048] 2. How to use the kit

[0049] The method for using the kit comprises the following steps:

[0050] 1. Extraction of viral RNA

[0051] ①Take 500 μL each of the sample to be tested, the positive control, and the negative control into 1.5ml sterilized EP tubes without RNase contamination, add an equal amount of Trizol lysate for lysing, shake fully, and place at room temperature for 10 minutes;

[0052...

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Abstract

The invention discloses an RT-PCR (Reverse Transcription-Polymerase Chain Reaction) primer for detecting European and American porcine reproductive and respiratory syndrome viruses and a kit. The nucleotide sequence of an upstream primer of the primer is 5'-ATGTGGGATAGGGTGGACA-3', and the nucleotide sequence of a downstream primer is 5'-AAGAACCAGAAAAGACACCCAA-3'; the kit comprises the primer. The European and American RT-PCR primer and the kit are adopted for detecting whether the European and American porcine reproductive and respiratory syndrome viruses are positive, and have the advantages of high sensitivity, strong specificity, fastness, convenience and low detection cost and the like.

Description

technical field [0001] The invention belongs to the technical field of animal virus molecular biological detection, in particular to a pair of RT-PCR primers and a kit for detecting the European type and the American type of porcine blue ear disease virus. Background technique [0002] Porcine reproductive and respiratory syndrome (PRRS) is an acute infectious disease caused by porcine reproductive and respiratory syndrome virus (PRRSV). The main clinical features are abortion, stillbirth, mummified fetus, and weak fetus. Difficulty breathing, etc. During the course of the disease, the skin of both ears will appear cyanotic and purple, so it is also called "blue ear disease". The disease was first reported in my country in 1996. [0003] According to the difference of PRRSV structural gene sequence, it can be divided into two genotypes of European type (represented by Lelystad Virus strain) and American type (represented by ATCC VR-232 strain). Among them, the American PRR...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68C12N15/11
CPCC12Q1/70C12Q1/686
Inventor 颜健华甘甲忠黄宇熊毅曾永芳冯淑萍李军张红云
Owner 广西壮族自治区动物疫病预防控制中心
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