Development of Reverse Transcription-Loop-Mediated Isothermal Amplification (RT-LAMP) Assay for Detection of Avian Influenza Viruses in Field Specimens

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  • SHIVAKOTI Sakar
    Laboratory of Veterinary Public Health, Faculty of Agriculture, Tottori University
  • ITO Hiroshi
    Laboratory of Veterinary Public Health, Faculty of Agriculture, Tottori University Avian Zoonosis Research Center, Faculty of Agriculture, Tottori University
  • MURASE Toshiyuki
    Avian Zoonosis Research Center, Faculty of Agriculture, Tottori University Laboratory of Veterinary Microbiology, Faculty of Agriculture, Tottori University
  • ONO Etsuro
    Avian Zoonosis Research Center, Faculty of Agriculture, Tottori University Laboratory of Biomedicine, Center of Biomedical Research, Graduate School of Medical Sciences, Kyushu University
  • TAKAKUWA Hiroki
    Avian Influenza Research Center, Kyoto Sangyo University
  • YAMASHIRO Tetsu
    Department of Virology, Institute of Tropical Medicine, Nagasaki University
  • OTSUKI Koichi
    Avian Zoonosis Research Center, Faculty of Agriculture, Tottori University Avian Influenza Research Center, Kyoto Sangyo University
  • ITO Toshihiro
    Laboratory of Veterinary Public Health, Faculty of Agriculture, Tottori University Avian Zoonosis Research Center, Faculty of Agriculture, Tottori University

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Abstract

Reverse transcription loop-mediated isothermal amplification (RT-LAMP) is an established gene amplification method for rapid diagnosis of various infectious diseases. In order to detect avian influenza viruses, particularly in field specimens, specific primers targeting the matrix gene were designed. Thirty-four virus samples, including isolates from wild and domestic avian hosts belonging to various geographical areas, were used to confirm the validity of the primers. All samples were confirmed to be positive in less than 1 hr. The RT-LAMP assay was also able to detect avian influenza virus in the various field samples, such as swabs, tissues, and feces. These results indicate that the developed RT-LAMP assay with uniquely designed primers is potentially useful in comprehensive avian influenza surveillance.<br>

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