A rapid pulsed-field gel electrophoresis method of genotyping Haemophilus parasuis isolates

Lett Appl Microbiol. 2011 Jun;52(6):589-95. doi: 10.1111/j.1472-765X.2011.03048.x. Epub 2011 Apr 21.

Abstract

Aims: To develop a modified pulsed-field gel electrophoresis (PFGE) method for characterizing Haemophilus parasuis isolates.

Methods and results: A modified PFGE procedure was designed using CpoI to generate restriction maps of H. parasuis genomic DNA. This approach was used to characterize 47 H. parasuis clinical isolates and 15 reference strains. All strains could be typed by this method, and the procedure was completed in 36 h. A total of 39 different PFGE patterns were identified among 47 epidemiologically unrelated clinical isolates.

Conclusions: The modified PGFE described in this report efficiently characterized H. parasuis isolates. This method can be adopted for studying the epidemiology of Glässer's disease outbreaks in addition to differentiating and classifying previously untypeable H. parasuis isolates.

Significance and impact of the study: The modified PFGE method described is a novel means of characterizing H. parasuis isolates. It is also a highly discriminatory molecular typing method (discriminatory index of 0.98) that can overcome the limitations of serotyping.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • China
  • DNA Restriction Enzymes / isolation & purification
  • Electrophoresis, Gel, Pulsed-Field / methods*
  • Genotype
  • Haemophilus Infections / epidemiology
  • Haemophilus Infections / microbiology*
  • Haemophilus parasuis / classification*
  • Haemophilus parasuis / genetics*
  • Molecular Typing
  • Serotyping

Substances

  • DNA Restriction Enzymes