Detection of changes in avian influenza genome fragments by multitemperature single-strand conformational polymorphism

Mol Cell Probes. 2008 Oct-Dec;22(5-6):301-4. doi: 10.1016/j.mcp.2008.09.003. Epub 2008 Oct 8.

Abstract

We have applied the temperature-dependent single-stranded conformational polymorphism (SSCP) analysis for characterization of influenza A virus cDNA fragments. A series of primers were synthesized on the basis of the comparison of hemagglutinin and PB2 gene sequences of different origin. RT-PCR reactions were run using these primers, products were denatured and single-strand DNA fragments were subjected to electrophoresis. After silver staining, ssDNA band patterns characteristic for each genotype were observed. Minor differences within a serotype were detected without need for sequencing the PCR products making the preliminary analysis of avian influenza isolates much faster.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Birds / virology
  • DNA, Complementary / genetics
  • Genome, Viral*
  • Genotype
  • Humans
  • Influenza A virus / genetics*
  • Influenza in Birds / classification
  • Influenza in Birds / diagnosis
  • Influenza in Birds / genetics
  • Phylogeny
  • Polymorphism, Single-Stranded Conformational*
  • Reverse Transcriptase Polymerase Chain Reaction / methods
  • Temperature

Substances

  • DNA, Complementary