Abstract:

【Objective】 The objective of the study is to establish a rapid detection of porcine parvovirus (PPV) by a loop-mediated isothermal amplification assay (LAMP). 【Method】 According to the published GenBank sequences, many pairs of primers were designed targeting the conserved region of PPV. The amplification was detected with LAMP Real Time Turbidimeter LA-320. Through optimizing the LAMP primers and reaction conditions, a rapid and specific detection of PPV was established. Meanwhile, the amplified products were colored by SYBR green I after completion of the reaction, so that the amplification could be detected with naked eyes. 【Result】 The method of LAMP shows a highly efficient amplification for PPV viral target gene which performed at 63 ℃ for 45min by the LAMP Real Time Turbidimeter LA-320. The detection limit was 0.23 TCID50. The results of PPV LAMP reaction visualized the tube directly with naked eyes by the addition of SYBR GreenⅠare consistent with the results detected by Tubidimeter real-time. Twenty clinical samples were detected by LAMP, PCR and IF, and the coincidence rate was 20/20. 【Conclusion】 PPV LAMP detection method established by the authors is rapid, specificity, high sensitivity, be easy of operation under simple conditions, which is suitable for rapid detection.

Key words: porcine parvovius, LAMP, detection