Sensitive detection of Renibacterium salmoninarum in whole fry, blood, and other tissues of pacific salmon by reverse transcription-polymerase chain reaction

Mol Mar Biol Biotechnol. 1998 Dec;7(4):270-9.

Abstract

A sensitive, reproducible assay for detecting Renibacterium salmoninarum in a variety of tissues, including blood, has been developed. This assay, based on reverse transcription-polymerase chain reaction (RT-PCR) of 16S ribosomal RNA, exhibited sensitivity to </=10 cells per milligram of tissue by ethidium bromide detection, and this sensitivity was increased 10-fold by Southern blotting. There was a strong association (p <.001) between blood and ovarian fluid for the presence of bacteria in spawning salmon, and in 17.9% of the infected fish, bacteria were detected in blood but not in ovarian fluid. The ability to analyze multiple tissues, the reproducibility, and sensitivity of 16S RT-PCR make it a useful tool for both research and husbandry applications.

Publication types

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

MeSH terms

  • Animals
  • Fish Diseases / diagnosis
  • Fish Diseases / microbiology*
  • Gram-Positive Bacteria / genetics
  • Gram-Positive Bacteria / isolation & purification*
  • Gram-Positive Bacterial Infections / diagnosis
  • Gram-Positive Bacterial Infections / microbiology
  • Gram-Positive Bacterial Infections / veterinary*
  • Kidney / microbiology
  • Kidney Diseases / diagnosis
  • Kidney Diseases / microbiology
  • Kidney Diseases / veterinary
  • RNA, Ribosomal, 16S / genetics
  • Reverse Transcriptase Polymerase Chain Reaction
  • Salmon / microbiology*
  • Sensitivity and Specificity

Substances

  • RNA, Ribosomal, 16S