Development of a loop-mediated isothermal amplification for visual detection of the HCLV vaccine against classical swine fever in China

Abstract

A loop-mediated isothermal amplification (LAMP) assay was developed and evaluated for the rapid and specific detection of HCLV vaccine strain against classical swine fever. Four primers were designed for amplification of NS5B gene region with Bst DNA polymerase at a constant temperature of 65 degrees C. The products showed ladder-like pattern on 2% agarose gel, and can be visualised after addition of SYBR Green I dye. The detection limit of the assay was 5 copies of the HCLV genome per reaction. No cross-reaction with other porcine viruses including different wild-type CSFV strains and the bovine viral diarrhoea virus was observed. The agreement between the LAMP and TaqMan real-time RT-PCR assays was 94.4% for the detection of 72 batches of HCLV vaccine. The assay provides a rapid tool for the control of vaccine quality and can be an accompanying assay of the LAMP for wild-type CSFV described previously for differential diagnosis. (C) 2010 Elsevier B.V. All rights reserved.

Keywords

Classical swine fever virus; Loop-mediated isothermal amplification; Hog cholera lapinized virus; Vaccine

Published in

Journal of Virological Methods
2011, Volume: 171, number: 1, pages: 200-205 Publisher: ELSEVIER SCIENCE BV

SLU Authors

• Belak, Sandor

  • Department of Animal Biosciences, Swedish University of Agricultural Sciences
    

• Liu, Lihong

  • Department of Animal Biosciences, Swedish University of Agricultural Sciences
    

UKÄ Subject classification

Biochemicals


Publication identifier

DOI: https://doi.org/10.1016/j.jviromet.2010.10.025

Permanent link to this page (URI)

https://res.slu.se/id/publ/47400