నైరూప్య

Primary development of lamp method for porcine circovirus type2 detection

Yang Zexiao, Zeng Hui, Yao Xueping, Wang Yin, Liu Bo, Wu Xulong, Peng Bin, Li Guili

To develop quickmethods for Porcine circovirus type2(PCV2) detection. 4 specific loop-mediated isothermal amplification (LAMP) primers were designed and synthesized acc- ording to the conserved sequences of ORF2 gene of porcine circovirus type 2(PCV2) publish- ed in GenBank, and the LAMP assay was established and evaluated by the preparation of tar- get gene fragments, optimization of reaction conditions, sensitivity and specificity tests. Then 6 strains PCV2 were detected using the LAMP. Results showthat the LAMP method for the PCV2 detection has a ladderlike pattern of amplification bands fromabout 134 bp performed at 64ºCfor 45min by using agarose gel electrophoresis, and with good sensitivity and specifi- city, the detection limitation reaches 10copy of target genomic fragments, and no amplifica- tion for PCV1, PRV, PPV, HCV and PRRV detection, and all of the 6 strains PCV2 prove to be positive by the LAMP method. This LAMP method established here provides a new useful, rapid, sensitive, reliable way to detect PCV2.