Detection of Francisella piscicida in Atlantic cod (Gadus morhua L) by the loop-mediated isothermal amplification (LAMP) reaction

Vet J. 2010 Jun;184(3):357-61. doi: 10.1016/j.tvjl.2009.03.027. Epub 2009 Apr 23.

Abstract

The loop-mediated isothermal amplification (LAMP) reaction was evaluated for its speed and sensitivity in detecting the presence of Francisella piscicida, the causative agent of francisellosis in Atlantic cod (Gadus morhua). Four primer sets, consisting of two outer and two inner, were designed from the groEL gene of the pathogen. The LAMP reaction was optimised at 63 degrees C for 1h using bacterial genomic DNA as the template and the products were visualised under ultra-violet light and analysed by agarose gel electrophoresis. A ladder-like pattern of bands, specific for F. piscicida, was amplified from positive samples. The method was highly specific for the detection of F. piscicida and was 100 times more sensitive than conventional PCR. In addition, the LAMP assay was able to detect the pathogen in kidney and splenic samples of naturally-infected Atlantic cod.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • DNA, Bacterial / analysis
  • Electrophoresis, Agar Gel / veterinary*
  • Fish Diseases / diagnosis*
  • Fish Diseases / microbiology
  • Francisella / genetics
  • Francisella / isolation & purification*
  • Gadus morhua / microbiology*
  • Gram-Negative Bacterial Infections / diagnosis
  • Gram-Negative Bacterial Infections / microbiology
  • Gram-Negative Bacterial Infections / veterinary*
  • Nucleic Acid Amplification Techniques / veterinary
  • Polymerase Chain Reaction / veterinary
  • Sensitivity and Specificity

Substances

  • DNA, Bacterial