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

Ghanem M. Al-Ghamdi, Vivek Kapur, Trevor R. Ames, John F. Timoney, Daria N. Love, Martha A. Mellencamp

Research output: Contribution to journalArticle

16 Citations (Scopus)

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 repPCR 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.

Original languageEnglish (US)
Pages (from-to)699-705
Number of pages7
JournalAmerican Journal of Veterinary Research
Volume61
Issue number6
DOIs
StatePublished - Jan 1 2000

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Streptococcus equi
Equus
repetitive sequences
Nucleic Acid Repetitive Sequences
polymerase chain reaction
Polymerase Chain Reaction
Disease Outbreaks
Horses
Clone Cells
Gels
DNA
clones
horses
Sepharose
Canada
Electrophoresis
genomics
agarose
gel electrophoresis
Population

All Science Journal Classification (ASJC) codes

  • veterinary(all)

Cite this

Al-Ghamdi, Ghanem M. ; Kapur, Vivek ; Ames, Trevor R. ; Timoney, John F. ; Love, Daria N. ; Mellencamp, Martha A. / Use of repetitive sequence-based polymerase chain reaction for molecular epidemiologic analysis of Streptococcus equi subspecies equi. In: American Journal of Veterinary Research. 2000 ; Vol. 61, No. 6. pp. 699-705.
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Use of repetitive sequence-based polymerase chain reaction for molecular epidemiologic analysis of Streptococcus equi subspecies equi. / Al-Ghamdi, Ghanem M.; Kapur, Vivek; Ames, Trevor R.; Timoney, John F.; Love, Daria N.; Mellencamp, Martha A.

In: American Journal of Veterinary Research, Vol. 61, No. 6, 01.01.2000, p. 699-705.

Research output: Contribution to journalArticle

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AU - Al-Ghamdi, Ghanem M.

AU - Kapur, Vivek

AU - Ames, Trevor R.

AU - Timoney, John F.

AU - Love, Daria N.

AU - Mellencamp, Martha A.

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AB - 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 repPCR 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.

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