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A novel RT-LAMP assay for rapid and simple detection of classical swine fever virus

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Virologica Sinica

Abstract

A simple and rapid assay for the detection of Classical swine fever virus (CSFV) was established using reverse transcription loop-mediated isothermal amplification (RT-LAMP). This study describes the amplification of the genomic RNA of CSFV under isothermal conditions (63°C) within one hour, using a set of six primers (two outer primers, two inner primers and two loop primers). This RT-LAMP assay showed 100-fold higher sensitivity than the standard RT-PCR method and identified eighteen additional positive cases that were negative when tested by RT-PCR. This RT-LAMP was able to detect all the 13 strains of CSFV but not the BVDV. PRRSV. SIV. PRV-PCV, thus showed a good specificity. Products amplified by RT-LAMP can be visualized by agarose gel electrophoresis and in addition, either as a white precipitate at the bottom of the tube after a pulse spin or as a color change when dyed with SYBR Green I which are visible to the naked eye. Because RT-LAMP is low-cost and produces rapid results, it has the potential to be an excellent tool for CSFV surveillance in the field, especially in developing countries.

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Authors and Affiliations

  1. China Institute of Veterinary Drug Control, Beijing, 100081, China

    Lei Chen, Xue-zheng Fan, Qin Wang, Lu Xu, Qi-zu Zhao, Yuan-chen Zhou, Jun Liu, Bo Tang & Xing-qi Zou

Authors
  1. Lei Chen
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  2. Xue-zheng Fan
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  3. Qin Wang
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  4. Lu Xu
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  5. Qi-zu Zhao
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  6. Yuan-chen Zhou
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  7. Jun Liu
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  8. Bo Tang
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  9. Xing-qi Zou
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Corresponding author

Correspondence to Qin Wang.

Additional information

Foundation items: The National Science and Technology supporting plan of the Eleventh Five-year (2006BAD 06A18 and 2006BAD06A03); Beijing Natural Science Foundation (5072041).

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Chen, L., Fan, Xz., Wang, Q. et al. A novel RT-LAMP assay for rapid and simple detection of classical swine fever virus. Virol. Sin. 25, 59–64 (2010). https://doi.org/10.1007/s12250-010-3043-2

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  • DOI: https://doi.org/10.1007/s12250-010-3043-2

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