Sammanfattning

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 re-ions 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 DNA was concentrated in the capture step. Furthermore, the use of the mimic molecule in the system assured that false negative results could be identified. (C) 2004 Elsevier B.V. All rights reserved

Nyckelord

Mimic; PCR; Renibacterium salmoniarum; sequence capture; 16S rRNA

Publicerad i

Veterinary Microbiology
2005, Volym: 105, nummer: 3-4, sidor: 235-243
Utgivare: ELSEVIER SCIENCE BV

SLU författare

• Johansson, Karl-Erik

  • Institutionen för husdjurens biovetenskaper, Sveriges lantbruksuniversitet
    

UKÄ forskningsämne

Husdjursvetenskap
Veterinärmedicin


Publikationens identifierare

DOI: https://doi.org/10.1016/j.vetmic.2004.11.007

Permanent länk till denna sida (URI)

https://res.slu.se/id/publ/7576