Abstract

Primary development of lamp method for porcine circovirus type2 detection

Author(s): 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.