Sensitive detection of Renibacterium salmoninarum in whole fry, blood, and other tissues of pacific salmon by reverse transcription-polymerase chain reaction.
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.
Rhodes
L D
LD
Resource Enhancement and Utilization Technologies Division, Northwest Fisheries Science Center, Seattle, Washington, 98112, USA.
Nilsson
W B
WB
Strom
M S
MS
D000818:Animals; D005393:Fish Diseases / Q000175:diagnosis / Q000382:microbiology*; D006094:Gram-Positive Bacteria / Q000235:genetics / Q000302:isolation & purification*; D016908:Gram-Positive Bacterial Infections / Q000175:diagnosis / Q000382:microbiology / Q000662:veterinary*; D007668:Kidney / Q000382:microbiology; D007674:Kidney Diseases / Q000175:diagnosis / Q000382:microbiology / Q000662:veterinary; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012474:Salmon / Q000382:microbiology*; D012680:Sensitivity and Specificity
D012336:RNA, Ribosomal, 16S