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. 2012 Jan;61(Pt 1):137-141.
doi: 10.1099/jmm.0.036046-0. Epub 2011 Sep 8.

Interlaboratory reproducibility of DiversiLab rep-PCR typing and clustering of Acinetobacter baumannii isolates

Paul G Higgins  1 , Andrea M Hujer  2   3 , Kristine M Hujer  2   3 , Robert A Bonomo  2   3   4   5 , Harald Seifert  1
Affiliations

Interlaboratory reproducibility of DiversiLab rep-PCR typing and clustering of Acinetobacter baumannii isolates

Paul G Higgins et al. J Med Microbiol. 2012 Jan.

Abstract

We have investigated the reproducibility of DiversiLab rep-PCR fingerprints between two laboratories with the aim of determining if the fingerprints and clustering are laboratory-specific or portable. One-hundred non-duplicate Acinetobacter baumannii isolates were used in this study. DNA isolation and rep-PCR were each performed separately in two laboratories and rep-PCR patterns generated in laboratory A were compared with those from laboratory B. Twelve A. baumannii isolates processed in laboratory A showed ≥98 % pattern similarity with the corresponding 12 isolates tested in laboratory B and were considered identical. Sixty-four isolates showed 95-97.9 % similarity with their corresponding isolates. Twenty-three isolates showed 90-94 % similarity with the corresponding isolates, while one isolate showed only 87.4 % similarity. However, intra-laboratory clustering was conserved: isolates that clustered in laboratory A also clustered in laboratory B. While clustering was conserved and reproducible at two different laboratories, demonstrating the robustness of rep-PCR, interlaboratory comparison of individual isolate fingerprints showed more variability. This comparison allows conclusions regarding clonality to be reached independent of the laboratory where the analysis is performed.

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Figures

Fig. 1.
Fig. 1.
Comparison of fingerprints using the PC and KL statistical methods. (a) Comparison between CL-17 (red line) and KO-17 (grey line). By the PC method the isolates show 96.6 % similarity and by the KL method 94.9 % similarity. (b) Comparison between CL-27 (red line) and KO-27 (grey line). By the PC method the isolates show 95 % similarity and by the KL method 90.9 % similarity.
Fig. 2.
Fig. 2.
Rep-PCR analysis. Dendrogram and computer-generated image of rep-PCR banding patterns showing clustering between fingerprints of corresponding isolate pairs generated in Cleveland (CL) and Cologne (KO) using the PC statistical method.
See this image and copyright information in PMC

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