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Development and validation of reverse transcription loop-mediated isothermal amplification for detection of PRRSV

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Abstract

To establish a reverse-transcription loop-mediated isothermal amplification (RT-LAMP) method for rapid detection of porcine reproductive and respiratory syndrome virus (PRRSV), four primers specific to six regions of the N gene were designed. After amplification in an isothermal water bath for 1 h, samples containing PRRSV generated the expected ladder-like products while porcine parvovirus, porcine circovirus, classic swine fever virus, pseudorabies virus, and swine testis cells generated no product. The sensitivity and specificity of the RT-LAMP assay were evaluated by comparison with reverse-transcription polymerase chain reaction (RT-PCR) and real-time PCR. Because it is specific and simple, the RT-LAMP assay will be useful for the diagnosis of PRRSV infection.

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Acknowledgments

The study was supported in part by funding from the National High-tech R&D Program (863 Program-2007AA100606) and the Chinese National Key Laboratory of Veterinary Biotechnology Fund (NKLVBP200807).

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Authors and Affiliations

  1. Division of Swine Infectious Disease, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of CAAS, No. 427, Maduan Street, Nangang District, Harbin, Heilongjiang, 150001, People’s Republic of China

    Changmu Chen, Shangjin Cui & Chaofan Zhang

  2. Institute of Animal Science and Veterinary Medicine, Shandong Academy of Agricultural Sciences, Jinan, 250100, China

    Jun Li & Jinbao Wang

Authors
  1. Changmu Chen
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  2. Shangjin Cui
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  3. Chaofan Zhang
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  4. Jun Li
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Correspondence to Shangjin Cui or Jinbao Wang.

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Chen, C., Cui, S., Zhang, C. et al. Development and validation of reverse transcription loop-mediated isothermal amplification for detection of PRRSV. Virus Genes 40, 76–83 (2010). https://doi.org/10.1007/s11262-009-0419-1

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  • DOI: https://doi.org/10.1007/s11262-009-0419-1

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