PCR-based fingerprinting of Mycobacterium bovis isolates.
We investigated a number of PCR-based strategies for sub-typing isolates of Mycobacterium bovis. A panel of 15 PCR primers, designed from random sequences, insertion sequences and repetitive elements, were analyzed for their ability to allow differentiation between eight M. bovis isolates. PCR products were analyzed using agarose gel electrophoresis and polyacrylamide gel electrophoresis. Random primer based PCR, conducted with a variety of primers did not differentiate between isolates of M. bovis. Successful differentiation between isolates was achieved by the amplification of fragments located between DNA repetitive elements and insertion sequences of the M. bovis genome. PCR primers designed from the major polymorphic tandem repeat (MPTR) region and from insertion sequences IS6110 and IS986 allowed differentiation between isolates of M. bovis. This study is presented as a first step towards the development of PCR-based methods to differentiate between isolates.
Glennon
M
M
National Diagnostics Centre, BioResearch Ireland, University College Galway, Ireland. maura.glennon@ucg.ic
Jäger
B
B
Dowdall
D
D
Maher
M
M
Dawson
M
M
Quigley
F
F
Costello
E
E
Smith
T
T
D000818:Animals; D001483:Base Sequence; D002417:Cattle; D016172:DNA Fingerprinting / Q000379:methods*; D017931:DNA Primers; D004269:DNA, Bacterial / Q000302:isolation & purification; D003670:Deer; D004587:Electrophoresis, Agar Gel; D004591:Electrophoresis, Polyacrylamide Gel; D005819:Genetic Markers; D006801:Humans; D009163:Mycobacterium bovis / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods; D011110:Polymorphism, Genetic
D017931:DNA Primers; D004269:DNA, Bacterial; D005819:Genetic Markers