Detection of Renibacterium salmoninarum in tissue samples by sequence capture and fluorescent PCR based on the 16S rRNA gene
2005 (English)In: Veterinary Microbiology, Vol. 105, no 3-4, 235-243 p.Article in journal (Refereed) Published
The 16S rRNA genes from eight isolates of Renibacterium salmoninarum with different origins and dates of isolation were
sequenced to evaluate the possibility to construct a diagnostic PCR system with target sites within this gene. The sequences were
found to be identical but for one single position in one of the isolates, and two regions with an adequate number of nucleotide
differences as compared to closely related species were identified. Species-specific fluorescent PCR primers complementary to
these regions were constructed as well as oligonucleotides for DNA preparation by sequence capture. A mimic molecule was
constructed to be used as an internal control. The PCR was specific and allowed the detection of DNA equivalent to 1–10 R.
salmoninarum genomes per reaction. The DNA preparation with sequence capture and analysis by PCR with a mimic was found
to be a reliable method for analysis of kidneys from fish with BKD. The amount of PCR inhibiting substances present in the
tissue was reduced, and the relevant DNAwas concentrated in the capture step. Furthermore, the use of the mimic molecule in
the system assured that false negative results could be identified.
Place, publisher, year, edition, pages
2005. Vol. 105, no 3-4, 235-243 p.
mimic, PCR, Renibacterium salmoninarum, sequence capture, 16S rRNA
IdentifiersURN: urn:nbn:se:uu:diva-70214OAI: oai:DiVA.org:uu-70214DiVA: diva2:98125