Specific detection of Mycobacterium paratuberculosis by DNA hybridisation with a fragment of the insertion element IS900.
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.
Moss
M T
MT
Department of Surgery, St George's Hospital Medical School, London.
Green
E P
EP
Tizard
M L
ML
Malik
Z P
ZP
Hermon-Taylor
J
J
D001483:Base Sequence; D003424:Crohn Disease / Q000235:genetics / Q000382:microbiology; D015342:DNA Probes / Q000235:genetics; D004251:DNA Transposable Elements; D006801:Humans; D007231:Infant, Newborn; D015336:Molecular Probe Techniques; D008969:Molecular Sequence Data; D009161:Mycobacterium / Q000235:genetics / Q000302:isolation & purification*; D009693:Nucleic Acid Hybridization; D010283:Paratuberculosis / Q000382:microbiology; D016133:Polymerase Chain Reaction
D015342:DNA Probes; D004251:DNA Transposable Elements