Rapid Detection of Foot-and-Mouth Disease Virus by Reverse Transcription Loop-mediated Isothermal Amplification (RT-LAMP)

Jun-Jun Shao; Hui-Yun Chang; Guang-Qing Zhou; Guo-Zheng Cong; Jun-Zheng Du; Tong Lin; Shan-Dian Gao; Ji-Jun He; Xiang-Tao Liu; Ji-Xing Liu; Jin-Liang Gao
June 2010
International Journal of Applied Research in Veterinary Medicine;Jun2010, Vol. 8 Issue 2, p133
Academic Journal
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, O, Asia1 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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