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

2010; Japanese Society of Veterinary Science; Volume: 72; Issue: 4 Linguagem: Inglês

10.1292/jvms.09-0473

1347-7439

Sakar Shivakoti, Hiroshi Ito, Toshiyuki Murase, Etsuro Ono, Hiroki Takakuwa, Tetsu Yamashiro, Koichi Otsuki, Toshihiro Ito,

Biosensors and Analytical Detection

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.