猪繁殖与呼吸综合征病毒RT-LAMP快速检测方法的建立

doi:10.16431/j.cnki.1671-7236.2017.06.008

Abstract

Abstract:

In order to diagnose porcine reproductive and respiratory syndrome virus (PRRSV) early, a rapid, sensitive, simple reverse transcription-loop-mediated isothermal amplification (RT-LAMP) assay was established. Four primers targeting the ORF5 gene of PRRSV were designed. Reverse transcription and amplification of the viral cDNA using Bst DNA polymerase was optimal at a constant temperature of 65 ℃. The output of the RT-LAMP assay was visualized using 1.0% agarose gel electrophoresis and color change after the addition of the SYBR Green Ⅰ dye. The assay was also specific for PRRSV and did not cross react with classical swine fever virus (CSFV), porcine circovirus virus type 2 (PCV2), porcine parvovirus (PPV), porcine pseudorabies virus (PRV). The RT-LAMP method was approximately 100-fold more sensitive than RT-PCR for PRRSV detection. The clinical samples (n=10) were identified both by RT-PCR and RT-LAMP, and the coincidece rate was 100%. Thus, the novel RT-LAMP assay was a rapid, simple, sensitive, specific test for PRRSV, and it could potentially be applied in clinical settings.

Key words: reverse transcription loop-mediated isothermal amplification (RT-LAMP); porcine reproductive and respiratory syndrome virus (PRRSV); rapid detection

CLC Number:

S852.65

CHEN Xi-wen, WANG Qian, YIN Miao, LI Lian, LUO Wen-tao, YANG Feng, GUO Ai-wei, ZHOU Jie-long, WANG Xiong-qing. Establishment of RT-LAMP for Rapid Detection of Porcine Reproductive and Respiratory Syndrome Virus[J]. , 2017, 44(6): 1630-1636.

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Establishment of RT-LAMP for Rapid Detection of Porcine Reproductive and Respiratory Syndrome Virus

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