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Journal of Clinical Microbiology, July 2009, p. 2226-2231, Vol. 47, No. 7
0095-1137/09/$08.00+0     doi:10.1128/JCM.02362-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Comparison of Two Multiple-Locus Variable-Number Tandem-Repeat Analysis Methods for Molecular Strain Typing of Human Brucella melitensis Isolates from the Middle East{triangledown}

Rebekah V. Tiller,1* Barun K. De,1 Marie Boshra,2 Lynn Y. Huynh,3 Matthew N. Van Ert,3 David M. Wagner,3 John Klena,2 T. S. Mohsen,4 S. S. El-Shafie,4 Paul Keim,3 Alex R. Hoffmaster,1 Patricia P. Wilkins,1 and Guillermo Pimentel2

U.S. Centers for Disease Control and Prevention, 1600 Clifton Road NE, MS-G34, Atlanta, Georgia 30333,1 U.S. Naval Medical Research Unit No. 3 (NAMRU-3), PSC 452, Box 5000, FPO AE 09835-0007, Cairo, Egypt,2 Northern Arizona University, Center for Microbial Genetics and Genomics, Flagstaff, Arizona 86011-5640,3 Hamad Medical Corporation, Doha, Qatar4

Received 9 December 2008/ Returned for modification 26 April 2009/ Accepted 6 May 2009

Brucella species are highly monomorphic, with minimal genetic variation among species, hindering the development of reliable subtyping tools for epidemiologic and phylogenetic analyses. Our objective was to compare two distinct multiple-locus variable-number tandem-repeat analysis (MLVA) subtyping methods on a collection of 101 Brucella melitensis isolates from sporadic human cases of brucellosis in Egypt (n = 83), Qatar (n = 17), and Libya (n = 1). A gel-based MLVA technique, MLVA-15IGM, was compared to an automated capillary electrophoresis-based method, MLVA-15NAU, with each MLVA scheme examining a unique set of variable-number tandem repeats. Both the MLVAIGM and MLVANAU methods were highly discriminatory, resolving 99 and 101 distinct genotypes, respectively, and were able to largely separate genotypes from Egypt and Qatar. The MLVA-15NAU scheme presented higher strain-to-strain diversity in our test population than that observed with the MLVA-15IGM assay. Both schemes were able to genetically correlate some strains originating from the same hospital or region within a country. In addition to comparing the genotyping abilities of these two schemes, we also compared the usability, limitations, and advantages of the two MLVA systems and their applications in the epidemiological genotyping of human B. melitensis strains.

* Corresponding author. Mailing address: U.S. Centers for Disease Control and Prevention, 1600 Clifton Road NE, MS-G34, Atlanta, GA 30333. Phone: (404) 639-4507. Fax: (404)-639-3023. E-mail: RVaughnTiller{at}cdc.gov

{triangledown} Published ahead of print on 13 May 2009.

Journal of Clinical Microbiology, July 2009, p. 2226-2231, Vol. 47, No. 7
0095-1137/09/$08.00+0     doi:10.1128/JCM.02362-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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