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Journal of Clinical Microbiology, January 2001, p. 94-100, Vol. 39, No. 1
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.1.94-100.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Detection and Differentiation of Old World Orthopoxviruses: Restriction Fragment Length Polymorphism of the crmB Gene Region

Vladimir N. Loparev,1,* Robert F. Massung,1 Joseph J. Esposito,1 and Hermann Meyer2

Division of Viral and Rickettsial Diseases, National Center for Infectious Diseases, Centers for Disease Control and Prevention, U.S. Department of Health and Human Services, Atlanta, Georgia 30333,1 and Federal Armed Forces Medical Academy, Institute of Microbiology, D-80937 Munich, Germany2

Received 12 May 2000/Returned for modification 21 August 2000/Accepted 23 October 2000

A restriction fragment length polymorphism (RFLP) assay was developed to identify and differentiate Old World, African-Eurasian orthopoxviruses (OPV): variola, vaccinia, cowpox, monkeypox, camelpox, ectromelia, and taterapox viruses. The test uses amplicons produced from virus genome DNA by PCR with a consensus primer pair designed from sequences determined for the cytokine response modifier B (crmB) gene of 43 different OPV strains of known taxonomic origin. The primer pair amplified a single specific product from each of the 115 OPV samples tested. Size-specific amplicons identified and differentiated ectromelia and vaccinia virus strains, which contain a truncated crmB gene, and enabled their differentiation from other OPV species. Restriction digests of amplified products allowed the identification and differentiation of variola, monkeypox, camelpox, vaccinia, and cowpox virus species and strains.

* Corresponding author. Mailing address: Division of Viral and Rickettsial Diseases, National Center for Infectious Diseases, CDC, MS D-10, 1600 Clifton Rd., Atlanta, GA 30333. Phone: (404) 639-4040. Fax: (404) 639-4056. E-mail: vnl0{at}cdc.gov.

Journal of Clinical Microbiology, January 2001, p. 94-100, Vol. 39, No. 1
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.1.94-100.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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Antimicrob. Agents Chemother. Clin. Microbiol. Rev.
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Copyright © 2001 by the American Society for Microbiology. All rights reserved.