Development of One-Step Reverse Transcription Loop-Mediated Isothermal Amplification (LAMP) as a Screening tool for Influenza A Viruses

Authors

  • Hein Min Tun
  • Trong Wisedchanwet
  • Manoosak Wongphatcharachai
  • Nutthawan Nonthabenjawan
  • Supasama Chaiyawong
  • Apiradee Theamboonlers
  • Yong Poovorawan
  • Alongkorn Amonsin

Keywords:

influenza A, reverse transcription loop-mediated isothermal amplification

Abstract

Influenza A virus is a major cause of influenza pandemics and can infect several host species including humans and animals. The objective of this study was to develop a one-step reverse transcription loop-mediated isothermal amplification assay (LAMP) for the detection of genetically diverse influenza A viruses from both human and animal hosts. First, a set of two inner and two outer primers were designed based on the conserved region of the matrix (M) gene of influenza A viruses. The amplification reaction was optimized at 63oC for 60 min and performed in a simple heat block. The amplicons could be visualized either by gel electrophoresis or by visual analysis upon addition of SybrGreen. The developed LAMP assay was tested with 50 influenza A isolates including H1N1, H1N2, H3N2, H5N1 and H7N4 from swine, avian and human hosts. In sensitivity test, the assay detection capability was ten times more sensitive than conventional RT-PCR and comparable to real time RT-PCR. In summary, this assay is a rapid, simple and sensitive assay suitable for less-equipped laboratories and thus can be utilized in the field as a screening test.

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Published

2014-12-15

How to Cite

Tun, H. M., Wisedchanwet, T., Wongphatcharachai, M., Nonthabenjawan, N., Chaiyawong, S., Theamboonlers, A., Poovorawan, Y., & Amonsin, A. (2014). Development of One-Step Reverse Transcription Loop-Mediated Isothermal Amplification (LAMP) as a Screening tool for Influenza A Viruses. The Thai Journal of Veterinary Medicine, 44(4), 427–434. Retrieved from https://he01.tci-thaijo.org/index.php/tjvm/article/view/25817

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Original Articles