Rapid Detection of Foot-and-Mouth Disease Virus by Reverse Transcription Loop-mediated Isothermal Amplification (RT-LAMP)
文献类型: 外文期刊
第一作者: Shao, Jun-Jun
作者: Shao, Jun-Jun;Chang, Hui-Yun;Zhou, Guang-Qing;Cong, Guo-Zheng;Du, Jun-Zheng;Lin, Tong;Gao, Shan-Dian;He, Ji-Jun;Liu, Xiang-Tao;Liu, Ji-Xing;Gao, Jin-Liang
作者机构:
关键词: Foot and mouth disease;transcription loop-medicated isothermal one-step single-tube method of method of reverse transcription loop-mediated isothermal polymerase amplification;RNA polymerase 3D(pol)
期刊名称:INTERNATIONAL JOURNAL OF APPLIED RESEARCH IN VETERINARY MEDICINE ( 影响因子:0.306; 五年影响因子:0.339 )
ISSN: 1542-2666
年卷期: 2010 年 8 卷 2 期
页码:
收录情况: SCI
摘要: The aim of this study was to produce a sensitive, simple and rapid diagnostic method for the detection of foot-and-mouth disease virus (FMDV) in suspected cases of FMD, a one-step single-tube method of reverse transcription loop-mediated isothermal polymerase amplification (RT-LAMP). A set of six common primers was designed to target the highly conserved region of the RNA polymerase 3Dpol gene of the seven FMDV serotypes. The sensitivity and specificity of RT-LAMP were evaluated by detection of 10-fold serial dilutions of the standard plasmids, and samples from experimental infection and suspected cases of FMD. The results showed that the target nucleic acid of four serotypes of FMDV (A, 0, Asia! and C) can be amplified rapidly by LAMP in a water bath in less than an hour. At least 10 copies of the DNA could be detected by RT-LAMP, which showed the same sensitivity as real-time PCR and another technique, RT-LAMP-1, but 10 times higher than that of reverse transcription polymerase chain reaction (RT-PCR). All 104 samples were detected by RT-LAMP, RT-LAMP-1, RT-PCR and real-time PCR; the positive ratios were 98.31%, 86.44%, 93.22%, and 100%, respectively. The results indicate that RT-LAMP is a rapid, cost-effective, efficient and simple method. Therefore it may be applied for the rapid detection of FMDV in the laboratory or under field conditions.
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