Use of repetitive sequence-based polymerase chain reaction for molecular epidemiologic analysis of Streptococcus equi subspecies equi

Ghanem M. Al-Ghamdi Departments of Clinical and Population Sciences, College of Veterinary Medicine, University of Minnesota, St Paul, MN 55108.

Search for other papers by Ghanem M. Al-Ghamdi in
Current site
Google Scholar
PubMed
Close
 DVM, MS
,
Vivek Kapur Department of Veterinary PathoBiology, College of Veterinary Medicine, University of Minnesota, St Paul, MN 55108.

Search for other papers by Vivek Kapur in
Current site
Google Scholar
PubMed
Close
 BVSc, PhD
,
Trevor R. Ames Departments of Clinical and Population Sciences, College of Veterinary Medicine, University of Minnesota, St Paul, MN 55108.

Search for other papers by Trevor R. Ames in
Current site
Google Scholar
PubMed
Close
 DVM, MS
,
John F. Timoney Department of Veterinary Science, College of Agriculture, University of Kentucky, Lexington, KY 40546.

Search for other papers by John F. Timoney in
Current site
Google Scholar
PubMed
Close
 PhD
,
Daria N. Love Department of Veterinary Anatomy and Pathology, Faculty of Veterinary Science, University of Sydney, Sydney, NSW 2006 Australia.

Search for other papers by Daria N. Love in
Current site
Google Scholar
PubMed
Close
 PhD
, and
Martha A. Mellencamp Departments of Clinical and Population Sciences, College of Veterinary Medicine, University of Minnesota, St Paul, MN 55108.

Search for other papers by Martha A. Mellencamp in
Current site
Google Scholar
PubMed
Close
 PhD

Abstract

Objective—To determine whether repetitive sequence-based polymerase chain reaction (rep-PCR) could be used to differentiate Streptococcus equi isolates, to examine S equi isolates from throughout the world, and to determine whether a horse had > 1 subtype of S equi during an outbreak of disease.

Sample Population—An initial group of 32 S equi isolates, 63 S equi isolates from various geographic areas, and 17 S equi isolates obtained during outbreaks of disease.

Procedure—An aliquot of S equi genomic DNA was amplified, using enterobacterial repetitive intergenic consensus primers. Gel electrophoresis was performed on 1.5% agarose gels, and a computed-assisted program was used to compare rep-PCR results.

Results—Use of these primers to analyze 100 ng of S equi genomic DNA resulted in patterns of 6 to 14 bands. The 32 initial isolates were separated into 7 rep- PCR subtypes. There were 30 rep-PCR subtypes found among 29 S equi isolates obtained from Minnesota, Michigan, Canada, and Australia and 34 S equi isolates obtained from Kentucky and other sources. Furthermore, the same clone was identified in several horses during an outbreak of disease. Infected horses on the same farm all had a single clone of S equi.

Conclusion and Clinical Relevance—Analysis of these results suggests that rep-PCR is useful for delineating S equi into rep-PCR subtypes. Results revealed that isolates with the same geographic source or similar date of collection did not always have the same rep-PCR subtype. A single clone of S equi usually predominated during an outbreak of disease. (Am J Vet Res 2000;61:699–705)

Abstract

Objective—To determine whether repetitive sequence-based polymerase chain reaction (rep-PCR) could be used to differentiate Streptococcus equi isolates, to examine S equi isolates from throughout the world, and to determine whether a horse had > 1 subtype of S equi during an outbreak of disease.

Sample Population—An initial group of 32 S equi isolates, 63 S equi isolates from various geographic areas, and 17 S equi isolates obtained during outbreaks of disease.

Procedure—An aliquot of S equi genomic DNA was amplified, using enterobacterial repetitive intergenic consensus primers. Gel electrophoresis was performed on 1.5% agarose gels, and a computed-assisted program was used to compare rep-PCR results.

Results—Use of these primers to analyze 100 ng of S equi genomic DNA resulted in patterns of 6 to 14 bands. The 32 initial isolates were separated into 7 rep- PCR subtypes. There were 30 rep-PCR subtypes found among 29 S equi isolates obtained from Minnesota, Michigan, Canada, and Australia and 34 S equi isolates obtained from Kentucky and other sources. Furthermore, the same clone was identified in several horses during an outbreak of disease. Infected horses on the same farm all had a single clone of S equi.

Conclusion and Clinical Relevance—Analysis of these results suggests that rep-PCR is useful for delineating S equi into rep-PCR subtypes. Results revealed that isolates with the same geographic source or similar date of collection did not always have the same rep-PCR subtype. A single clone of S equi usually predominated during an outbreak of disease. (Am J Vet Res 2000;61:699–705)

All Time Past Year Past 30 Days
Abstract Views 47 0 0
Full Text Views 1488 1242 23
PDF Downloads 90 42 9
Advertisement