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Specific detection of Mycobacterium paratuberculosis by DNA hybridisation with a fragment of the insertion element IS900.
  1. M T Moss,
  2. E P Green,
  3. M L Tizard,
  4. Z P Malik,
  5. J Hermon-Taylor
  1. Department of Surgery, St George's Hospital Medical School, London.

    Abstract

    This paper describes the evaluation of a newly developed DNA probe for Mycobacterium paratuberculosis. DNA probe PCR278 is a 278 bp fragment obtained by polymerase chain reaction (PCR) amplification of the 5'-region of IS900, an insertion element contained in the genome of M paratuberculosis. This DNA probe can specifically distinguish M paratuberculosis from a wide range of other organisms, including members of the M avium-M intracellulare complex. When used in conjunction with the PCR amplification technique DNA probe PCR278 could detect as little as 10 fg (equivalent to two genomes) starting material of M paratuberculosis genomic DNA. Use of PCR amplification assays based on IS900, for the detection of M paratuberculosis, and homologous IS elements found in disease isolates of M avium should greatly help our understanding of the role of these organisms in Crohn's disease and other chronic inflammatory disorders.

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