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Primer probe group and kit for detecting type-2 porcine reproductive and respiratory syndrome virus by one-step method

A technology for respiratory syndrome and primer probes, which is applied in recombinant DNA technology, microbial assay/inspection, biochemical equipment and methods, etc.

Inactive Publication Date: 2020-10-20
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The prior art (Liu Yongsheng et al., ZL201310375408.2) reported a conventional PCR diagnostic method for type 1 and type 2 PRRSV, but it is difficult to meet the requirements of large-throughput detection in actual detection, and it is relatively time-consuming and laborious. In practice, there will be false positives due to the level of operators

Method used

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  • Primer probe group and kit for detecting type-2 porcine reproductive and respiratory syndrome virus by one-step method
  • Primer probe group and kit for detecting type-2 porcine reproductive and respiratory syndrome virus by one-step method
  • Primer probe group and kit for detecting type-2 porcine reproductive and respiratory syndrome virus by one-step method

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Embodiment 1

[0035] 1. Sample nucleic acid extraction

[0036] Nucleic acids were extracted from various samples using the MiniBEST Viral RNA / DNA Extraction Kit produced by Takara Bioengineering (Dalian) Co., Ltd.

[0037] 2. Routine RT-PCR reaction experiment to determine PRRSV

[0038] 1. Primers used in multiplex RT-PCR reactions

[0039] The primers used in the multiplex RT-PCR reaction are the 4 primers described in patent ZL201310375408.2, the first pair of primers are: 5'-CAC CACGTCGAAAGTGCCGC-3'(SEQ ID NO.5) and 5'-GACGCCGGACGACAAATGCG-3'(SEQ ID NO. .6); the second pair of primers is: 5'-GGGGCTGTTGCACATCCTAA-3' (SEQ ID NO.7) and 5'-ACAAAAGGGCAACAACAGCC-3' (SEQ ID NO.8), for the amplification of type 1 and type 2 PRRSV The sizes are approximately 120bp and 70bp, respectively.

[0040] 2. Multiplex RT-PCR amplification reaction system and amplification reaction process

[0041] Amplification conditions described in patent ZL201310375408.2: 50°C for 30 minutes, 94°C for 2 minutes,...

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Abstract

The invention relates to a primer probe group and a kit for detecting a type-2 porcine reproductive and respiratory syndrome virus by a one-step method, and belongs to the technical field of virus detection. The probe primer group comprises a primer F1, a primer F2, a primer R and a probe, wherein the nucleotide sequence of the primer F1 is disclosed by SEQ ID NO.1, the nucleotide sequence of theprimer F2 is disclosed by SEQ ID NO.2, the nucleotide sequence of the primer R is disclosed by SEQ ID NO.3, and the probe is a substance that the 5' terminal of the nucleotide sequence disclosed by the SEQ ID NO.4 is modified with a fluorophore and the 3' terminal is modified with a quenching group. When the primer probe group disclosed by the invention is applied to detection, specificity and sensitivity are good, the primer probe group does not react with the type-1 porcine reproductive and respiratory syndrome, and a clinical application value is high.

Description

technical field [0001] The invention relates to the technical field of virus detection, in particular to a primer probe set and a kit for one-step detection of type 2 porcine reproductive and respiratory syndrome virus. Background technique [0002] Porcine reproductive and respiratory syndrome virus (Porcine reproductive and respiratory syndrome virus, PRRSV) is the pathogen of porcine reproductive and respiratory syndrome (Porcine reproductive and respiratory syndrome, PRRS). A highly contagious disease of pigs that causes dyspnea in finishing pigs. According to antigenic and genomic characteristics, PRRSV can be divided into two genotypes: European type (European, Eu), that is, type 1 PRRSV; North American type (North American, NA), that is, type 2 PRRSV. Prevailing in my country is type 2 PRRSV. [0003] PRRS disease has always been one of the most important diseases that plague the healthy development of my country's pig industry. Due to the lack of serological metho...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/6851C12N15/11
CPCC12Q1/701C12Q1/6851C12Q2600/166C12Q2531/113C12Q2563/107C12Q2545/113
Inventor 丁耀忠张杰张中旺马维民何继军刘永生陈豪泰
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI