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Comparative Study
. 2012 Jan;32(1):50-6.
doi: 10.3343/alm.2012.32.1.50. Epub 2011 Dec 20.

Comparison of modified multiple-locus variable-number tandem-repeat fingerprinting with pulsed-field gel electrophoresis for typing clinical isolates of Staphylococcus aureus

Soie Chung  1 , Jongyoun Yi, Mi Hee Jang, Sei-Ick Joo, Eun Kyung Ra, So Yeon Kim, Chulhun L Chang, Sung Sup Park, Eui-Chong Kim
Affiliations
Comparative Study

Comparison of modified multiple-locus variable-number tandem-repeat fingerprinting with pulsed-field gel electrophoresis for typing clinical isolates of Staphylococcus aureus

Soie Chung et al. Ann Lab Med. 2012 Jan.

Abstract

Background: Multiple-locus variable-number tandem-repeat fingerprinting (MLVF) is based on multiplex PCR, utilizing variable number tandem repeat. Our goal was to compare the performance of MLVF in distinguishing clinical Staphylococcus aureus isolates with that of pulsed-field gel electrophoresis (PFGE), which has traditionally been the gold standard.

Methods: Sixty-three clinically significant S. aureus isolates were tested using both PFGE and MLVF. Multiplex PCR for MLVF was performed using PCR primers for clfA, clfB, sdrCDE, sspA, and spa. PFGE was performed with genomic DNA fragments generated by SmaI endonuclease digestion. Banding patterns of MLVF or PFGE were analyzed using InfoQuestFP software.

Results: The hands-on time of our modified method was about 3 h, on average, for each of 18 isolates. PFGE (80% cutoff) or MLVF (75% cutoff) separated all of the 63 isolates into 13 and 12 types, respectively. Three types generated by PFGE were identical to those generated by MLVF. PFGE and MLVF yielded similar Simpson's diversity indices, indicating similar discriminatory power. The overall concordance between PFGE and MLVF was low, as represented by adjusted Rand indices (0.266-0.278). PFGE predicted MLVF type better than MLVF predicted PFGE type, as reflected by Wallace coefficients (PFGE cutoff 80% vs. MLVF cutoff 75%, 0.389 vs. 0.233). Analysis of the relationship between a pair of isolates showed 91.0% concordance between the PFGE (80% cutoff) and MLVF (75% cutoff).

Conclusions: Our simple, low-cost, modified MLVF protocol can effectively discriminate between S. aureus clinical isolates. MLVF can replace PFGE for the hospital infection control of S. aureus.

Keywords: Multiple-locus variable-number tandem-repeat fingerprinting; Pulsed-field gel electrophoresis; Staphylococcus aureus.

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Conflict of interest statement

No potential conflict of interest relevant to this article was reported.

Figures

Fig. 1
Fig. 1
Pulsed-field gel electrophoresis (PFGE) types of the study isolates generated by the unweighted pair-group method with arithmetic mean (UPGMA) algorithm, using 80% and 75% similarity cutoff values. The isolates that were not included in the above types were considered to be unique patterns. Isolate numbers presented in boldface were not included in any types generated by the 80% cutoff.
Fig. 2
Fig. 2
Multiple-locus variable-number tandem-repeat fingerprinting (MLVF) types of the study isolates generated by the unweighted pairgroup method with arithmetic mean (UPGMA) algorithm using 75% and 70% similarity cutoff values. The isolates that were not included in the above types were considered to be unique patterns. Isolate numbers presented in boldface were not included in any types generated by the 75% cutoff.
See this image and copyright information in PMC

References

    1. Wertheim HF, Vos MC, Ott A, van Belkum A, Voss A, Kluytmans JA, et al. Risk and outcome of nosocomial Staphylococcus aureus bacteraemia in nasal carriers versus non-carriers. Lancet. 2004;364:703–705. - PubMed
    1. Wertheim HF, van Leeuwen WB, Snijders S, Vos MC, Voss A, Vandenbroucke-Grauls CM, et al. Associations between Staphylococcus aureus genotype, infection, and in-hospital mortality: a nested case-control study. J Infect Dis. 2005;192:1196–1200. - PubMed
    1. Cosgrove SE. The relationship between antimicrobial resistance and patient outcomes: mortality, length of hospital stay, and health care costs. Clin Infect Dis. 2006;42(Suppl 2):S82–S89. - PubMed
    1. Struelens MJ, Deplano A, Godard C, Maes N, Serruys E. Epidemiologic typing and delineation of genetic relatedness of methicillin-resistant Staphylococcus aureus by macrorestriction analysis of genomic DNA by using pulsed-field gel electrophoresis. J Clin Microbiol. 1992;30:2599–2605. - PMC - PubMed
    1. Tenover FC, Vaughn RR, McDougal LK, Fosheim GE, McGowan JE., Jr Multiple-locus variable-number tandem-repeat assay analysis of methicillin-resistant Staphylococcus aureus strains. J Clin Microbiol. 2007;45:2215–2219. - PMC - PubMed

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