Rapid detection of porcine kobuvirus in feces by reverse transcription loop-mediated isothermal amplification
1 Division of Swine Infectious Diseases, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of the Chinese Academy of Agricultural Sciences, No.427 Maduan Street, Nangang District, Harbin 150001, China
2 College of Life Science, Northeast Agricultural University, Harbin 150030, China
Virology Journal 2014, 11:73 doi:10.1186/1743-422X-11-73Published: 23 April 2014
PKV is a new emerging pathogen detected in diarrhea pigs. At present, no more detection methods were reported except RT-PCR method. this study was to develop a fast diagnostic method based on the LAMP reaction for rapid detection of PKV nucleic acid in fecal samples.
Two pairs of primers were designed to amplify the conservative 3D gene of PKV genome. The PKV RT-LAMP method possessed well specificity and had 100 times higher sensitivity than common reverse transcription PCR (RT-PCR), which could detect up to 10 RNA copies of the target gene.
The results showed that the optimal reaction condition for RT-LAMP was achieved at 64°C for 50 min. Furthermore, the RT-LAMP procedure does not demand special equipment and is time-saving.