Detection of swine transmissible gastroenteritis coronavirus using loop-mediated isothermal amplification | Chen et al. Virology Journal 2010 7 206 http content 7 1 206 VIROLOGY JOURNAL RESEARCH Open Access Detection of swine transmissible gastroenteritis coronavirus using loop-mediated isothermal amplification Qin Chen1 Jian Li2 Xue-En Fang1 Wei Xiong2 Abstract A conserved nucleic acid fragment of the nucleocapsid gene of Swine Transmissible Gastroenteritis Coronavirus TGEV was chosen as the target six special primers were designed successfully. Loop-mediated isothermal amplification LAMP was developed to detect the TGEV by incubation at 60 C for 1 h and the product specificity was confirmed by HphI digestion. Standard curves with high accuracy for TGEV quantization was constructed by adding 1 X SYBR greenl in the LAMP reaction. The assay established in this study was found to detect only the TGEV and no cross-reaction with other viruses demonstrating its high specificity. By using serial sample dilutions as templates the detection limit of LAMP was about 10 pg RNA 10 times more sensitive than that of PCR and could be comparable to the nest-PCR. Background Swine Transmissible Gastroenteritis Coronavirus TGEV as a member of the coronaviridae is a kind of single-stranded RNA virus which produces villous atrophy and enteritis leading to the serious financial loss to the whole pig industry. The traditional detection methods including virus isolation virus immunodiagnostic assays and PCR tests have the shortcomings such as precise instruments requirement elaborate result analysis demand high cost long detection time and so forth which prevent these methods from being widely used 1-4 . Loop-mediated isothermal amplification LAMP is a novel nucleic acid amplification method which amplifies DNA RNA with high specificity sensitivity and rapidity under isothermal condition 5 . It has already found wide application in RNA virus detection such as Foot-and-mouth Disease Virus 6 Swine Vesicular Disease Virus 7 Taura Syndrome Virus 8 Severe Acute Respiratory Syndrome
