猪呼吸与繁殖综合症病毒RT-LAMP检测方法的建立

doi:10.3864/j.issn.0578-1752.2010.01.022

Scientia Agricultura Sinica ›› 2010, Vol. 43 ›› Issue (1): 185-191 .doi: 10.3864/j.issn.0578-1752.2010.01.022

• VETERINARY SCIENCE • Previous Articles Next Articles

Development of a Novel RT-LAMP Assay for Porcine Reproductive and Respiratory Syndrome Virus

XIN Ting, HOU Shao-hua, JIA Hong, GUO Xiao-yu, DING Jia-bo, LI Yan-peng, DING Min, ZHU Hong-fei

（中国农业科学院北京畜牧兽医研究所）

Received:2008-11-14 Revised:2009-10-10 Online:2010-01-10 Published:2010-01-10
Contact: ZHU Hong-fei

Abstract

Abstract:

【Objective】 The objective of the experiment is to develop a one-step reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for rapid and sensitive detection of porcine reproductive and respiratory syndrome virus (PRRSV). 【Method】 Three pairs of primers were designed to identify 8 positions in N gene, the sensitivity and specificity of RT-LAMP were determined and clinical test was performed under optimized amplification condition. 【Result】 A RT-LAMP assay was developed for the rapid detection of PRRSV. The process of assay was conducted with one step amplification within 70 minutes and amplification results were visualized. The sensitivity of RT-LAMP assay was 10 000 times higher than RT-PCR. In the detection of 50 porcine serum samples, this assay showed excellent agreement with the standard RT-PCR assay (κ＝0.83). 【Conclusion】 Therefore, the rapid and simple assay is a potential useful technique for PRRSV detection in field condition.

Key words: PRRSV, RT-LAMP, detection

XIN Ting,HOU Shao-hua,JIA Hong,GUO Xiao-yu,DING Jia-bo,LI Yan-peng,DING Min,ZHU Hong-fei
. Development of a Novel RT-LAMP Assay for Porcine Reproductive and Respiratory Syndrome Virus
[J].Scientia Agricultura Sinica, 2010, 43(1): 185-191 .

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