Verification of an Edwardsiella ictaluri-specific diagnostic PCR

Lett Appl Microbiol. 2010 Feb;50(2):153-7. doi: 10.1111/j.1472-765X.2009.02770.x. Epub 2009 Nov 6.

Abstract

Aims: To verify the specificity of a PCR assay for the identification and diagnosis of Edwardsiella ictaluri.

Methods and results: An Edwardsiella ictaluri-specific PCR assay was developed utilizing two features of the ribosomal DNA gene clusters. The first feature is the presence of two ribosomal gene clusters located in tandem to one another (the inter-ribosomal spacer, IRS). This characteristic is present in the Edwardsiella genus but absent in the other sequenced members of the Enterobacteriaceae. The second feature is the presence of an intervening sequence (IVS) in the 23S rRNA gene of Edw. ictaluri. To verify the specificity of this assay, we tested genomic DNA from a variety of bacterial species. The IVS/IRS PCR assay results in an c. 2000-bp product from all Edw. ictaluri isolates tested, but not from any other species including Edwardsiella tarda.

Conclusions: The IVS/IRS PCR assay is highly specific for Edw. ictaluri and useful as a tool for identifying this pathogen.

Significance and impact of the study: This research verifies the specificity of PCR-based assay for Edw. Ictaluri, and we describe this assay as a highly versatile diagnostic tool for its identification.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Catfishes / microbiology
  • DNA, Bacterial / genetics*
  • DNA, Ribosomal Spacer / genetics*
  • Edwardsiella ictaluri / genetics*
  • Edwardsiella ictaluri / isolation & purification
  • Edwardsiella tarda / genetics
  • Fish Diseases / diagnosis
  • Fish Diseases / genetics
  • Fish Diseases / microbiology
  • Multigene Family / genetics*
  • Polymerase Chain Reaction / methods*
  • RNA, Bacterial / genetics
  • RNA, Ribosomal, 23S / genetics
  • Sensitivity and Specificity
  • Species Specificity

Substances

  • DNA, Bacterial
  • DNA, Ribosomal Spacer
  • RNA, Bacterial
  • RNA, Ribosomal, 23S