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Primers, kit and method for detecting porcine reproductive and respiratory syndrome virus

A technique of syndromic virus and porcine respiration, applied in the field of molecular biology, to achieve the effect of saving time and cost, high specificity and sensitivity, and simple operation

Active Publication Date: 2019-09-17
WENS FOODSTUFF GRP CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Timely and accurate diagnosis is very important for the prevention and control of diseases. The NADC30-like strain is a new epidemic strain, and there is no PCR diagnostic method for this type of strain at present

Method used

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  • Primers, kit and method for detecting porcine reproductive and respiratory syndrome virus
  • Primers, kit and method for detecting porcine reproductive and respiratory syndrome virus
  • Primers, kit and method for detecting porcine reproductive and respiratory syndrome virus

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Embodiment 1

[0025] Example 1 Primers and methods for detecting porcine respiratory and reproductive disorder syndrome virus

[0026] 1, Primer design

[0027] According to the porcine respiratory and reproductive disorder syndrome virus sequence registered by NCBI, accession numbers: mutant strains (EU864233, EF112445, FJ548853), classic strains (EU864233, EF536003, AF066183) and NADC30-like strains (JN654459, KP861625) provided Information, after comparative analysis, designed PCR primers, respectively F, R:

[0028] Upstream primer F: ACCTCCTTTGAYTGGRATGTTGTGT (SEQ ID NO: 1);

[0029] Downstream primer R: CTTGAGCYGAGTAYTTYGGGCG (SEQ ID NO: 2)

[0030] (Merged base Y stands for C or T; R stands for A or G)

[0031] Detection method

[0032] Include the following steps:

[0033] (1) RNA extraction from samples to be tested

[0034] 200 μL of cell culture medium (infected with the self-isolated strain ZW, isolated from the lung of the diseased pig, belonging to the self-isolated ...

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Abstract

The invention discloses a primer, kit and method for detecting porcine reproductive and respiratory syndrome viruses. The primer is as shown in SEQ ID No.1 and SEQ ID No.2. The method includes: using the RNA of a to-be-detected sample as the template, performing RT-PCR amplification reaction, and performing electrophoresis identification. The primer, kit and method has the advantages that the primer is high in specificity and sensitivity and capable of detecting the novel NADC30 viruses of the porcine reproductive and respiratory syndrome; PCR amplification can fast identify the classical strains, variants and strains similar to NADC30 of the PRRSV (porcine reproductive and respiratory syndrome viruses) through the size differences of amplification fragments, the sequencing step is omitted, time and cost are saved, and simpleness in operation and practicality are achieved.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, and more specifically, the invention relates to a primer, a kit and a method for detecting porcine reproductive and respiratory syndrome virus. Background technique [0002] Pig blue-ear disease is caused by porcine reproductive and respiratory syndrome virus (Porcine Reproductive and Respiratory Syndrome Virus, PRRSV) infection. It is an infectious disease with reproductive disorders in sows and respiratory failure in piglets as the main symptoms. Abortion, premature birth, stillbirth, mummified fetus, decreased survival rate of piglets and respiratory symptoms of piglets. The disease first appeared in the United States in 1987 and has since spread rapidly around the world. [0003] Porcine Reproductive and Respiratory Syndrome (PRRS) has had a great impact on the pig industry, especially in North America and Europe in recent years, showing a trend of global spread, mainly with sympto...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/70C12Q1/686C12N15/11
Inventor 王东东宋延华潘永飞李春梅叶敏慧欧阳海平蔡新斌卢围
Owner WENS FOODSTUFF GRP CO LTD