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http://dx.doi.org/10.7853/kjvs.2015.38.3.145

Visual detection of porcine circovirus 2 by loop-mediated isothermal amplification (LAMP) with hydroxynaphthol blue dye  

Kong, Ho-Chul (College of Veterinary Medicine & Animal Disease Intervention Center, Kyungpook National University)
Kim, Eun-Mi (College of Veterinary Medicine & Animal Disease Intervention Center, Kyungpook National University)
Jeon, Hyo-Sung (M monitor Incorporation)
Kim, Ji-Jung (M monitor Incorporation)
Kim, Hee-Jung (College of Veterinary Medicine & Animal Disease Intervention Center, Kyungpook National University)
Park, Yu-Ri (College of Veterinary Medicine & Animal Disease Intervention Center, Kyungpook National University)
Kang, Dae-Young (College of Veterinary Medicine & Animal Disease Intervention Center, Kyungpook National University)
Kim, Young-Hwa (National Institute of Animal Science, RDA)
Park, Jun-Cheol (National Institute of Animal Science, RDA)
Lee, Chang-hee (Department of Microbiology, College of Natural Sciences, Kyungpook National University)
Yeo, Sang-Geon (College of Veterinary Medicine & Animal Disease Intervention Center, Kyungpook National University)
Park, Choi-Kyu (College of Veterinary Medicine & Animal Disease Intervention Center, Kyungpook National University)
Publication Information
Korean Journal of Veterinary Service / v.38, no.3, 2015 , pp. 145-153 More about this Journal
Abstract
In this study, we developed a loop-mediated isothermal amplification (LAMP) with hydroxynaphtol blue dye (HNB) for rapid and direct visual detection of porcine circovirus 2 DNA with high sensitivity and specificity. The LAMP was completed in 40 min at $63^{\circ}C$, and the results of the LAMP can be confirmed by naked eye without any detection process. The sensitivity of the LAMP was 10-fold higher than that of the commercial PCR (cPCR) and real time PCR (rPCR) previously reported. In clinical application, the PCV2 detection rate of the LAMP was the same on porcine tissue samples (75.0%, 36/48) between porcine blood samples (75.0%, 39/52). The PCV2 detection rate (75.0%) of LAMP was higher than those of the cPCR and rPCR (67.3%, 35/52) in blood samples. In conclusion, the LAMP developed in the study could be an useful alternative method for the detection of PCV2 in the swine disease diagnostic laboratories.
Keywords
Porcine circovirus 2 (PCV2); loop-mediated isothermal amplification (LAMP); polymerase chain reaction (PCR); real time-PCR (rPCR);
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