Loop-mediated amplification of the Clavibacter michiganensis subsp. michiganensis micA gene is highly specific

Phytopathology. 2013 Dec;103(12):1220-6. doi: 10.1094/PHYTO-03-13-0078-R.

Abstract

Loop-mediated amplification (LAMP) was used to specifically identify Clavibacter michiganensis subsp. michiganensis, causal agent of bacterial canker of tomato. LAMP primers were developed to detect micA, a chromosomally stable gene that encodes a type II lantibiotic, michiganin A, which inhibits growth of other C. michiganensis subspecies. In all, 409 bacterial strains (351 C. michiganensis subsp. michiganensis and 58 non-C. michiganensis subsp. michiganensis) from a worldwide collection were tested with LAMP to determine its specificity. LAMP results were compared with genetic profiles established using polymerase chain reaction (PCR) amplification of seven genes (dnaA, ppaJ, pat-1, chpC, tomA, ppaA, and ppaC). C. michiganensis subsp. michiganensis strains produced eight distinct profiles. The LAMP reaction identified all C. michiganensis subsp. michiganensis strains and discriminated them from other C. michiganensis subspecies and non-Clavibacter bacteria. LAMP has advantages over immunodiagnostic and other molecular detection methods because of its specificity and isothermal nature, which allows for easy field application. The LAMP reaction is also not affected by as many inhibitors as PCR. This diagnostic tool has potential to provide an easy, one-step test for rapid identification of C. michiganensis subsp. michiganensis.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Base Sequence
  • DNA Primers / genetics
  • DNA, Bacterial / genetics
  • Genes, Bacterial / genetics*
  • Micrococcaceae / classification
  • Micrococcaceae / genetics
  • Micrococcaceae / isolation & purification*
  • Micrococcaceae / pathogenicity
  • Molecular Sequence Data
  • Phylogeny
  • Plant Diseases / microbiology*
  • Polymerase Chain Reaction / methods*
  • Sensitivity and Specificity
  • Sequence Analysis, DNA
  • Solanum lycopersicum / microbiology*
  • Species Specificity

Substances

  • DNA Primers
  • DNA, Bacterial

Associated data

  • GENBANK/KC416011
  • GENBANK/KC416012
  • GENBANK/KC416013
  • GENBANK/KC416014
  • GENBANK/KC416015
  • GENBANK/KC416016
  • GENBANK/KC416017
  • GENBANK/KC416018
  • GENBANK/KC416019
  • GENBANK/KC416020
  • GENBANK/KC416021
  • GENBANK/KC416022
  • GENBANK/KC416023
  • GENBANK/KC416024
  • GENBANK/KC416025