Virus Genes

, Volume 32, Issue 3, pp 261–267

Rapid Detection of Avian Influenza Virus A and Subtype H5N1 by Single Step Multiplex Reverse Transcription-polymerase Chain Reaction

  • Hui-Ling Wei
  • Gui-Rong Bai
  • Aaron S. Mweene
  • Ying-Chun Zhou
  • Yan-Long Cong
  • Juan Pu
  • Shuai Wang
  • Hiroshi Kida
  • Jin-Hua Liu
Original Paper

DOI: 10.1007/s11262-005-6910-4

Cite this article as:
Wei, HL., Bai, GR., Mweene, A.S. et al. Virus Genes (2006) 32: 261. doi:10.1007/s11262-005-6910-4

Abstract

Outbreaks of H5N1 highly pathogenic avian influenza (HPAI) virus caused great economic losses to the poultry industry and resulted in human deaths in Thailand and Viet Nam in 2004. Rapid typing and subtyping of H5N1 viruses, especially from clinical specimens, are desirable for taking prompt control measures to prevent the spread of the disease. Here, we developed a set of oligonucleotide primers able to detect, type and subtype H5 and N1 influenza viruses in a single step multiplex reverse transcription-polymerase chain reaction (RT-PCR). RNA was extracted from allantoic fluid or from specimens with guanidinium isothiocyanate reagent. Reverse transcription and PCR were carried out with a mixture of primers specific for influenza viruses of type A, subtype H5 and N1 in a single reaction system under identical conditions. The amplified DNA fragments were analyzed by agarose gel electrophoresis. All the H5N1 viruses tested in the study and the experimental specimens presented three specific bands by the method established here. The results presented here suggest that the method described below is rapid and specific and, therefore, could be valuable in the rapid detection of H5N1 influenza viruses in clinics.

Keywords

Avain influenza virusH5N1TypingSubtypingMultiplex RT-PCR

Copyright information

© Springer Science+Business Media, Inc. 2006

Authors and Affiliations

  • Hui-Ling Wei
    • 1
  • Gui-Rong Bai
    • 2
  • Aaron S. Mweene
    • 2
  • Ying-Chun Zhou
    • 1
  • Yan-Long Cong
    • 1
  • Juan Pu
    • 1
  • Shuai Wang
    • 1
  • Hiroshi Kida
    • 2
  • Jin-Hua Liu
    • 1
  1. 1.Department of Infectious DiseaseCollege of Veterinary Medicine, China Agricultural UniversityBeijingPeople’s Republic of China
  2. 2.Laboratory of Microbiology, Department of Disease ControlHokkaido University Graduate School of Veterinary MedicineSapporoJapan