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Journal of Clinical Microbiology, October 1999, p. 3217-3222, Vol. 37, No. 10
Central Veterinary Research Laboratory,
Abbotstown, Castleknock, Dublin 15,1 and
National Agricultural and Veterinary Biotechnology
Centre2 and Veterinary Epidemiology and
Tuberculosis Investigation Unit,3 University
College Dublin, Dublin 4, Ireland
Received 7 December 1998/Returned for modification 2 April
1999/Accepted 7 April 1999
Restriction fragment length polymorphism (RFLP) analysis with
probes derived from the insertion element IS6110, the
direct repeat sequence, and the polymorphic GC-rich sequence (PGRS) and a PCR-based typing method called spacer oligonucleotide typing (spoligotyping) were used to strain type Mycobacterium
bovis isolates from the Republic of Ireland. Results were
assessed for 452 isolates which were obtained from 233 cattle, 173 badgers, 33 deer, 7 pigs, 5 sheep, and 1 goat. Eighty-five strains were
identified by RFLP analysis, and 20 strains were identified by
spoligotyping. Twenty percent of the isolates were the most prevalent
RFLP type, while 52% of the isolates were the most prevalent
spoligotype. Both the prevalent RFLP type and the prevalent spoligotype
were identified in isolates from all animal species tested and had a
wide geographic distribution. Isolates of some RFLP types and some
spoligotypes were clustered in regions consisting of groups of
adjoining counties. The PGRS probe gave better differentiation of
strains than the IS6110 or DR probes. The majority of
isolates from all species carried a single IS6110 copy. In
four RFLP types IS6110 polymorphism was associated with
deletion of fragments equivalent in size to one or two direct variable
repeat sequences. The same range and geographic distribution of strains
were found for the majority of isolates from cattle, badgers, and deer.
This suggests that transmission of infection between these species is a
factor in the epidemiology of M. bovis infection in Ireland.
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Study of Restriction Fragment Length Polymorphism
Analysis and Spoligotyping for Epidemiological Investigation of
Mycobacterium bovis Infection
*
Corresponding author. Mailing address: Eamon Costello,
Central Veterinary Research Laboratory, Abbotstown, Castleknock, Dublin 15, Ireland. Phone: (353) 1 6072789. Fax: (353) 1 8213010. E-mail: eamoncos{at}indigo.ie.
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