Limits...
Molecular epidemiology of clinical isolates of Pseudomonas aeruginosa isolated from horses in Ireland.

Tazumi A, Maeda Y, Buckley T, Millar B, Goldsmith C, Dooley J, Elborn J, Matsuda M, Moore J - Ir Vet J (2009)

Bottom Line: Resulting banding patterns (n = 24 genotypes) showed a high degree of genetic heterogeneity amongst all isolates examined, indicating a relative non-clonal relationship between isolates from these patients, employing this genotyping technique.This study characterised 63 clinical isolates into 24 distinct genotypes, with the largest cluster (genotype E) accounting for 10/63 (15.9%) of the isolates.With the adoption of several controls to aid reproducibility, this technique may be useful as an alternative to PFGE, particularly in epidemiological investigations of outbreaks where speed may be a significant parameter.

View Article: PubMed Central - HTML - PubMed

Affiliation: Northern Ireland Public Health Laboratory, Department of Bacteriology, Belfast City Hospital, Belfast BT9 7AD, Northern Ireland. jemoore@niphl.dnet.co.uk.

ABSTRACT
Clinical isolates (n = 63) of Pseudomonas aeruginosa obtained from various sites in 63 horses were compared using ERIC2 RAPD PCR to determine their genetic relatedness. Resulting banding patterns (n = 24 genotypes) showed a high degree of genetic heterogeneity amongst all isolates examined, indicating a relative non-clonal relationship between isolates from these patients, employing this genotyping technique. This study characterised 63 clinical isolates into 24 distinct genotypes, with the largest cluster (genotype E) accounting for 10/63 (15.9%) of the isolates. ERIC2 RAPD PCR proved to be a highly discriminatory molecular typing tool of P. aeruginosa in isolates recovered from horses. With the adoption of several controls to aid reproducibility, this technique may be useful as an alternative to PFGE, particularly in epidemiological investigations of outbreaks where speed may be a significant parameter. This is the first report of clonal heterogeneity amongst P. aeruginosa from horses and demonstrated that ERIC RAPD PCR is a rapid method for the examination of this species in horses, which may be useful in outbreak analysis.

No MeSH data available.


Representative banding profiles associated with ERIC2 RAPD PCR in P. aeruginosa isolated from horses. Lane M: Molecular weight marker (100 bp); lane 1: genotype A; lanes 2 and 3, genotype B; lanes 4, 5 and 9, genotype C; lane 6, genotype D; lanes 7 and 13, genotype E; lane 8, genotype F; lane 10, genotype G; lane 11, genotype H; lane 12, genotype H; lane 14, genotype J; lane 15, genotype K; lane 16, genotype L; lane 17, genotype M; lane 18, genotype N.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC3113754&req=5

Figure 1: Representative banding profiles associated with ERIC2 RAPD PCR in P. aeruginosa isolated from horses. Lane M: Molecular weight marker (100 bp); lane 1: genotype A; lanes 2 and 3, genotype B; lanes 4, 5 and 9, genotype C; lane 6, genotype D; lanes 7 and 13, genotype E; lane 8, genotype F; lane 10, genotype G; lane 11, genotype H; lane 12, genotype H; lane 14, genotype J; lane 15, genotype K; lane 16, genotype L; lane 17, genotype M; lane 18, genotype N.

Mentions: All P. aeruginosa isolates examined in this study generated an ERIC2 RAPD banding pattern ranging in size, from approximately 200 bp to >1,000 bp, with between three and eight bands per isolate, with a mean band number of 5.3 bands per isolate examined (Figure 1). Overall, there was a high degree of clonal heterogeneity between all isolates examined, as 24 distinct RAPD genotypes were recorded (Genotype A-Genotype X) from the 63 isolates examined. Clustering of isolates was observed, where Genotype E was the most frequent cluster with 10 (10/63; 15.9% of total isolates) members, followed by Genotype R (8/63; 12.7%), Genotype C (6/63; 9.5%), Genotype B (5/63; 7.9%) and Genotype V (5/63; 7.0%). These five largest clusters accounted for 34/63 isolates (69.8%) examined in the study. Of the remaining genotypes observed, there were three genotypes each containing three isolates, five genotypes each containing two members and 11 genotypes represented by a single isolate. Certain genotypes with larger numbers of isolates had isolates from several years, i.e., for genotype E with 10 members, these isolates were representative from 2005-2007, whilst for genotype R, isolates here represented 2003-2006. Thus, this study demonstrated the persistence and recurrence of certain genotypes over a four year period within the equine population.


Molecular epidemiology of clinical isolates of Pseudomonas aeruginosa isolated from horses in Ireland.

Tazumi A, Maeda Y, Buckley T, Millar B, Goldsmith C, Dooley J, Elborn J, Matsuda M, Moore J - Ir Vet J (2009)

Representative banding profiles associated with ERIC2 RAPD PCR in P. aeruginosa isolated from horses. Lane M: Molecular weight marker (100 bp); lane 1: genotype A; lanes 2 and 3, genotype B; lanes 4, 5 and 9, genotype C; lane 6, genotype D; lanes 7 and 13, genotype E; lane 8, genotype F; lane 10, genotype G; lane 11, genotype H; lane 12, genotype H; lane 14, genotype J; lane 15, genotype K; lane 16, genotype L; lane 17, genotype M; lane 18, genotype N.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3113754&req=5

Figure 1: Representative banding profiles associated with ERIC2 RAPD PCR in P. aeruginosa isolated from horses. Lane M: Molecular weight marker (100 bp); lane 1: genotype A; lanes 2 and 3, genotype B; lanes 4, 5 and 9, genotype C; lane 6, genotype D; lanes 7 and 13, genotype E; lane 8, genotype F; lane 10, genotype G; lane 11, genotype H; lane 12, genotype H; lane 14, genotype J; lane 15, genotype K; lane 16, genotype L; lane 17, genotype M; lane 18, genotype N.
Mentions: All P. aeruginosa isolates examined in this study generated an ERIC2 RAPD banding pattern ranging in size, from approximately 200 bp to >1,000 bp, with between three and eight bands per isolate, with a mean band number of 5.3 bands per isolate examined (Figure 1). Overall, there was a high degree of clonal heterogeneity between all isolates examined, as 24 distinct RAPD genotypes were recorded (Genotype A-Genotype X) from the 63 isolates examined. Clustering of isolates was observed, where Genotype E was the most frequent cluster with 10 (10/63; 15.9% of total isolates) members, followed by Genotype R (8/63; 12.7%), Genotype C (6/63; 9.5%), Genotype B (5/63; 7.9%) and Genotype V (5/63; 7.0%). These five largest clusters accounted for 34/63 isolates (69.8%) examined in the study. Of the remaining genotypes observed, there were three genotypes each containing three isolates, five genotypes each containing two members and 11 genotypes represented by a single isolate. Certain genotypes with larger numbers of isolates had isolates from several years, i.e., for genotype E with 10 members, these isolates were representative from 2005-2007, whilst for genotype R, isolates here represented 2003-2006. Thus, this study demonstrated the persistence and recurrence of certain genotypes over a four year period within the equine population.

Bottom Line: Resulting banding patterns (n = 24 genotypes) showed a high degree of genetic heterogeneity amongst all isolates examined, indicating a relative non-clonal relationship between isolates from these patients, employing this genotyping technique.This study characterised 63 clinical isolates into 24 distinct genotypes, with the largest cluster (genotype E) accounting for 10/63 (15.9%) of the isolates.With the adoption of several controls to aid reproducibility, this technique may be useful as an alternative to PFGE, particularly in epidemiological investigations of outbreaks where speed may be a significant parameter.

View Article: PubMed Central - HTML - PubMed

Affiliation: Northern Ireland Public Health Laboratory, Department of Bacteriology, Belfast City Hospital, Belfast BT9 7AD, Northern Ireland. jemoore@niphl.dnet.co.uk.

ABSTRACT
Clinical isolates (n = 63) of Pseudomonas aeruginosa obtained from various sites in 63 horses were compared using ERIC2 RAPD PCR to determine their genetic relatedness. Resulting banding patterns (n = 24 genotypes) showed a high degree of genetic heterogeneity amongst all isolates examined, indicating a relative non-clonal relationship between isolates from these patients, employing this genotyping technique. This study characterised 63 clinical isolates into 24 distinct genotypes, with the largest cluster (genotype E) accounting for 10/63 (15.9%) of the isolates. ERIC2 RAPD PCR proved to be a highly discriminatory molecular typing tool of P. aeruginosa in isolates recovered from horses. With the adoption of several controls to aid reproducibility, this technique may be useful as an alternative to PFGE, particularly in epidemiological investigations of outbreaks where speed may be a significant parameter. This is the first report of clonal heterogeneity amongst P. aeruginosa from horses and demonstrated that ERIC RAPD PCR is a rapid method for the examination of this species in horses, which may be useful in outbreak analysis.

No MeSH data available.