Journal of Clinical Microbiology, April 2002, p. 1164-1173, Vol. 40, No. 4
0095-1137/02/$04.00+0 DOI: 10.1128/JCM.40.4.1164-1173.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.
Genotyping of a Homogeneous Group of Yersinia pestis Strains Isolated in the United States
Xiao-Zhe Huang,1 May C. Chu,2 Dave M. Engelthaler,2 and Luther E. Lindler1*
Department of Bacterial Diseases, Division of Communicable Diseases and Immunology, Walter Reed Army Institute of Research, Silver Spring, Maryland,1
Division of Vector-Borne Infectious Diseases, National Center for Infectious Diseases, Centers for Diseases Control and Prevention, Ft. Collins, Colorado2
Received 14 December 2001/
Accepted 6 January 2002
Yersinia pestis, the causative agent of deadly plague, is considered a reemerging infectious disease and a significant biological terrorism threat. The present project focused on epidemiological investigation of the genetic variability of well-documented strains of Y. pestis from the United States by pulsed-field gel electrophoresis (PFGE) and restriction fragment length polymorphism (RFLP) analysis with insertion sequences IS100 and IS285 as probes. We examined 37 U.S. Y. pestis strains and isolates of a single ribotype, ribotype B, recovered between 1939 and 1998 from patients, animals, and fleas. Our results showed that all isolates had similar PFGE patterns, but minor differences such as missing, additional, and shifted bands were found among almost all strains if they came from different parent strains. The 37 strains and isolates were divided into 26 PFGE types. RFLP analysis with IS100 as a probe divided these strains and isolates into 16 types, with 43% belonging to IS100 type 1. Typing with IS285 as a probe was less specific and led to only four RFLP types, with 81% belonging to type 1. Similarity analysis with BioNumerics software showed that all strains shared
80, 86, and 91% similarities on dendrograms prepared from digitized PFGE, IS100 RFLP analysis, and IS285 RFLP analysis images, respectively. Our results demonstrate that PFGE offers an increased ability to discriminate between strains (Simpson's index of diversity, 0.98) and therefore can significantly improve epidemiological studies related to the origin of new plague isolates.
* Corresponding author. Mailing address: Department of Bacterial Diseases, Division of Communicable Diseases and Immunology, WRAIR, 503 Robert Grant Ave., Silver Spring, MD 20910. Phone: (301) 319-9388. Fax: (301) 319-9123. E-mail: luther.lindler{at}NA.AMEDD.ARMY.MIL.
Journal of Clinical Microbiology, April 2002, p. 1164-1173, Vol. 40, No. 4
0095-1137/02/$04.00+0 DOI: 10.1128/JCM.40.4.1164-1173.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.
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Copyright © 2002 by the American Society for Microbiology. All rights reserved.