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Journal of Clinical Microbiology, October 1999, p. 3265-3270, Vol. 37, No. 10
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Diphtheria in the Republic of Georgia: Use of Molecular Typing Techniques for Characterization of Corynebacterium diphtheriae Strains

Alexander Sulakvelidze,1,* Merab Kekelidze,2 Tsaro Gomelauri,2 Yingkang Deng,1 Nino Khetsuriani,3,4 Ketino Kobaidze,5 Aruni De Zoysa,6 Androulla Efstratiou,6 J. Glenn Morris Jr.,1 and Paata Imnadze2

Division of Hospital Epidemiology, University of Maryland School of Medicine, Baltimore, Maryland 212011; National Center for Diseases Control of the Georgian Ministry of Health, Tbilisi 380077, Republic of Georgia2; National Immunization Program,3 Epidemic Intelligence Service of the Epidemiology Program Office,4 and National Center for Infectious Diseases,5 Centers for Disease Control and Prevention, Atlanta, Georgia 30333; and PHLS Streptococcus and Diphtheria Reference Unit of the Respiratory and Systemic Infection Laboratory, Central Public Health Laboratory, London NW9 5HT, England6

Received 8 April 1999/Returned for modification 17 May 1999/Accepted 24 June 1999

Sixty-six Corynebacterium diphtheriae strains (62 of the gravis biotype and 4 of the mitis biotype) isolated during the Georgian diphtheria epidemic of 1993 to 1998 and 13 non-Georgian C. diphtheriae strains (10 Russian and 3 reference isolates) were characterized by (i) biotyping, (ii) toxigenicity testing with the Elek assay and PCR, (iii) the randomly amplified polymorphic DNA (RAPD) technique, and (iv) pulsed-field gel electrophoresis (PFGE). Fifteen selected strains were ribotyped. Six RAPD types and 15 PFGE patterns were identified among all strains examined, and 12 ribotypes were found among the 15 strains that were ribotyped. The Georgian epidemic apparently was caused by one major clonal group of C. diphtheriae (PFGE type A, ribotype R1), which was identical to the predominant epidemic strain(s) isolated during the concurrent diphtheria epidemic in Russia. A dendrogram based on the PFGE patterns revealed profound differences between the minor (nonpredominant) epidemic strains found in Georgia and Russia. The methodologies for RAPD typing, ribotyping, and PFGE typing of C. diphtheriae strains were improved to enable rapid and convenient molecular typing of the strains. The RAPD technique was adequate for biotype differentiation; however, PFGE and ribotyping were better (and equal to each other) at discriminating between epidemiologically related and unrelated isolates.

* Corresponding author. Mailing address: Division of Hospital Epidemiology, University of Maryland School of Medicine, MSTF Bldg., Room 9-34, 10 S. Pine St., Baltimore, MD 21201. Phone: (410) 706-4587. Fax: (410) 706-4581. E-mail: asulakve{at}umppa1.ab.umd.edu.

Journal of Clinical Microbiology, October 1999, p. 3265-3270, Vol. 37, No. 10
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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