Preparation of RT-PCR Kits for Identification of Avian Influnza Virus Type A and Subtype H5
DOI:
Author:
Affiliation:

Clc Number:

S854.4

Fund Project:

  • Article
  • |
  • Figures
  • |
  • Metrics
  • |
  • Reference
  • |
  • Related
  • |
  • Cited by
  • |
  • Materials
  • |
  • Comments
    Abstract:

    Objective To develop RT-PCR kits for detection and identification of type A and subtype H5 avien influnza virus (AIV). Methods Two pairs of primes were designed for type A and subtype H5 avian influenza vir us according to the report by Ming-Shiuh Lee. The cDNA of AIV were amplified by one step RT-PCR for AIV type A and subtype H5, and identified by electrophores is analysis. The RT-PCT kits for AIV type A and subtype H5 were assembled. The H1-H15 reference strains and 38 isolated strains were detected using the RT -PCR kits. Results The RT-PCR method for detection of AIV type A and subt ype H5 was established. All stains of AIV were positive using RT-PCR kits for A type. The sensitivity of this kit was 1/1024 hemagglutinin unit. Subtype H5 reference strains and 19 subtype H5 isolated strains were positively detected using the RT-PCR kit for AIV subty pe H5. Subtypes H1-H4 and H6-H15 reference strains, subtypes H7 and H9 isolated strains, and 1 subtype H5 isolated strain showed a negative result. The sensiti vity of this kit was 1/64 hemagglutinin unit for AIV subtype H5. The samples of H5N1 AIV infected chicken were all positively detected using these two types of kits. Conclusion The RT-PCR kits for AIV type A and subtype H5 are excellent in specificity, sensitivity, stability and reproducibility.

    Reference
    Related
    Cited by
Get Citation
Share
Article Metrics
  • Abstract:
  • PDF:
  • HTML:
  • Cited by:
History
  • Received:
  • Revised:October 09,2004
  • Adopted:
  • Online:
  • Published: