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Detection of Renibacterium salmoninarum in tissue samples by sequence capture and fluorescent PCR based on the 16S rRNA gene
Uppsala University, Teknisk-naturvetenskapliga vetenskapsområdet, Chemistry, Surface Biotechnology. Uppsala University, Teknisk-naturvetenskapliga vetenskapsområdet, Chemistry, Department of Physical and Analytical Chemistry.
2005 (English)In: Veterinary Microbiology, Vol. 105, no 3-4, 235-243 p.Article in journal (Refereed) Published
Abstract [en]

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.
Keyword [en]
mimic, PCR, Renibacterium salmoninarum, sequence capture, 16S rRNA
National Category
URN: urn:nbn:se:uu:diva-70214OAI: oai:DiVA.org:uu-70214DiVA: diva2:98125
Available from: 2007-02-14 Created: 2007-02-14 Last updated: 2011-01-12

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Surface BiotechnologyDepartment of Physical and Analytical Chemistry

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