This Article
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Ropp, S. L.
Right arrow Articles by Esposito, J. J.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Ropp, S. L.
Right arrow Articles by Esposito, J. J.

 Previous Article  |  Next Article 

Journal of Clinical Microbiology, Aug 1995, 2069-2076, Vol 33, No. 8
Copyright © 1995 by the American Society for Microbiology. All rights reserved.

PCR strategy for identification and differentiation of small pox and other orthopoxviruses

SL Ropp, Q Jin, JC Knight, RF Massung and JJ Esposito
Division of Viral and Rickettsial Disease, Centers for Disease Control and Prevention, Atlanta, Georgia 30333, USA.

Rapid identification and differentiation of orthopoxviruses by PCR were achieved with primers based on genome sequences encoding the hemagglutinin (HA) protein, an infected-cell membrane antigen that distinguishes orthopoxviruses from other poxvirus genera. The initial identification step used a primer pair of consensus sequences for amplifying an HA DNA fragment from the three known North American orthopoxviruses (raccoonpox, skunkpox, and volepox viruses), and a second pair for amplifying virtually the entire HA open reading frame of the Eurasian-African orthopoxviruses (variola, vaccinia, cowpox, monkeypox, camelpox, ectromelia, and gerbilpox viruses). RsaI digest electropherograms of the amplified DNAs of the former subgroup provided species differentiation, and TaqI digests differentiated the Eurasian- African orthopoxviruses, including vaccinia virus from the vaccinia virus subspecies buffalopox virus. Endonuclease HhaI digest patterns distinguished smallpox variola major viruses from alastrim variola minor viruses. For the Eurasian-African orthopoxviruses, a confirmatory step that used a set of higher-sequence-homology primers was developed to provide sensitivity to discern individual virus HA DNAs from cross- contaminated orthopoxvirus DNA samples; TaqI and HhaI digestions of the individual amplified HA DNAs confirmed virus identity. Finally, a set of primers and modified PCR conditions were developed on the basis of base sequence differences within the HA genes of the 10 species, which enabled production of a single DNA fragment of a particular size that indicated the specific species.


This article has been cited by other articles:

  • Balamurugan, V., Bhanuprakash, V., Hosamani, M., Jayappa, K. D., Venkatesan, G., Chauhan, B., Singh, R. K. (2009). A polymerase chain reaction strategy for the diagnosis of camelpox. jvdi 21: 231-237 [Abstract] [Full Text]  
  • Bielinska, A. U., Chepurnov, A. A., Landers, J. J., Janczak, K. W., Chepurnova, T. S., Luker, G. D., Baker, J. R. Jr. (2008). A Novel, Killed-Virus Nasal Vaccinia Virus Vaccine. CVI 15: 348-358 [Abstract] [Full Text]  
  • Kurth, A., Achenbach, J., Miller, L., Mackay, I. M., Pauli, G., Nitsche, A. (2008). Orthopoxvirus Detection in Environmental Specimens during Suspected Bioterror Attacks: Inhibitory Influences of Common Household Products. Appl. Environ. Microbiol. 74: 32-37 [Abstract] [Full Text]  
  • HUTSON, C. L., LEE, K. N., ABEL, J., CARROLL, D. S., MONTGOMERY, J. M., OLSON, V. A., LI, Y., DAVIDSON, W., HUGHES, C., DILLON, M., SPURLOCK, P., KAZMIERCZAK, J. J., AUSTIN, C., MISER, L., SORHAGE, F. E., HOWELL, J., DAVIS, J. P., REYNOLDS, M. G., BRADEN, Z., KAREM, K. L., DAMON, I. K., REGNERY, R. L. (2007). MONKEYPOX ZOONOTIC ASSOCIATIONS: INSIGHTS FROM LABORATORY EVALUATION OF ANIMALS ASSOCIATED WITH THE MULTI-STATE US OUTBREAK. Am J Trop Med Hyg 76: 757-768 [Abstract] [Full Text]  
  • Scaramozzino, N., Ferrier-Rembert, A., Favier, A.-l., Rothlisberger, C., Richard, S., Crance, J.-M., Meyer, H., Garin, D. (2007). Real-Time PCR to Identify Variola Virus or Other Human Pathogenic Orthopox Viruses. Clin. Chem. 53: 606-613 [Abstract] [Full Text]  
  • Fedele, C. G., Negredo, A., Molero, F., Sanchez-Seco, M. P., Tenorio, A. (2006). Use of Internally Controlled Real-Time Genome Amplification for Detection of Variola Virus and Other Orthopoxviruses Infecting Humans. J. Clin. Microbiol. 44: 4464-4470 [Abstract] [Full Text]  
  • TRINDADE, G. S., LOBATO, Z. I. P., DRUMOND, B. P., LEITE, J. A., TRIGUEIRO, R. C., GUEDES, M. I. M. C., DA FONSECA, F. G., DOS SANTOS, J. R., BONJARDIM, C. A., FERREIRA, P. C. P., KROON, E. G. (2006). ISOLATION OF TWO VACCINIA VIRUS STRAINS FROM A SINGLE BOVINE VACCINIA OUTBREAK IN RURAL AREA FROM BRAZIL: IMPLICATIONS ON THE EMERGENCE OF ZOONOTIC ORTHOPOXVIRUSES.. Am J Trop Med Hyg 75: 486-490 [Abstract] [Full Text]  
  • Trindade, G. S., da Fonseca, F. G., Marques, J. T., Diniz, S., Leite, J. A., De Bodt, S., Van der Peer, Y., Bonjardim, C. A., Ferreira, P. C. P., Kroon, E. G. (2004). Belo Horizonte virus: a vaccinia-like virus lacking the A-type inclusion body gene isolated from infected mice. J. Gen. Virol. 85: 2015-2021 [Abstract] [Full Text]  
  • Olson, V. A., Laue, T., Laker, M. T., Babkin, I. V., Drosten, C., Shchelkunov, S. N., Niedrig, M., Damon, I. K., Meyer, H. (2004). Real-Time PCR System for Detection of Orthopoxviruses and Simultaneous Identification of Smallpox Virus. J. Clin. Microbiol. 42: 1940-1946 [Abstract] [Full Text]  
  • Panning, M., Asper, M., Kramme, S., Schmitz, H., Drosten, C. (2004). Rapid Detection and Differentiation of Human Pathogenic Orthopox Viruses by a Fluorescence Resonance Energy Transfer Real-Time PCR Assay. Clin. Chem. 50: 702-708 [Abstract] [Full Text]  
  • Guharoy, R., Panzik, R., Noviasky, J. A, Krenzelok, E. P, Blair, D. C (2004). Smallpox: Clinical Features, Prevention, and Management. The Annals of Pharmacotherapy 38: 440-447 [Abstract] [Full Text]  
  • Kulesh, D. A., Baker, R. O., Loveless, B. M., Norwood, D., Zwiers, S. H., Mucker, E., Hartmann, C., Herrera, R., Miller, D., Christensen, D., Wasieloski, L. P. Jr., Huggins, J., Jahrling, P. B. (2004). Smallpox and pan-Orthopox Virus Detection by Real-Time 3'-Minor Groove Binder TaqMan Assays on the Roche LightCycler and the Cepheid Smart Cycler Platforms. J. Clin. Microbiol. 42: 601-609 [Abstract] [Full Text]  
  • Reed, K. D., Melski, J. W., Graham, M. B., Regnery, R. L., Sotir, M. J., Wegner, M. V., Kazmierczak, J. J., Stratman, E. J., Li, Y., Fairley, J. A., Swain, G. R., Olson, V. A., Sargent, E. K., Kehl, S. C., Frace, M. A., Kline, R., Foldy, S. L., Davis, J. P., Damon, I. K. (2004). The Detection of Monkeypox in Humans in the Western Hemisphere. NEJM 350: 342-350 [Abstract] [Full Text]  
  • Sofi Ibrahim, M., Kulesh, D. A., Saleh, S. S., Damon, I. K., Esposito, J. J., Schmaljohn, A. L., Jahrling, P. B. (2003). Real-Time PCR Assay To Detect Smallpox Virus. J. Clin. Microbiol. 41: 3835-3839 [Abstract] [Full Text]  
  • Kak, V. (2003). Cowpox--Not Cows but Cats. Arch Intern Med 163: 249-249 [Full Text]  
  • Marik, P. E., Bowles, S. (2002). Analytic Reviews: Management of Patients Exposed to Biological and Chemical Warfare Agents. J Intensive Care Med 17: 147-161 [Abstract]  
  • Espy, M. J., Cockerill III, F. R., Meyer, R. F., Bowen, M. D., Poland, G. A., Hadfield, T. L., Smith, T. F. (2002). Detection of Smallpox Virus DNA by LightCycler PCR. J. Clin. Microbiol. 40: 1985-1988 [Abstract] [Full Text]  
  • Breman, J. G., Henderson, D.A. (2002). Diagnosis and Management of Smallpox. NEJM 346: 1300-1308 [Full Text]  
  • Lapa, S., Mikheev, M., Shchelkunov, S., Mikhailovich, V., Sobolev, A., Blinov, V., Babkin, I., Guskov, A., Sokunova, E., Zasedatelev, A., Sandakhchiev, L., Mirzabekov, A. (2002). Species-Level Identification of Orthopoxviruses with an Oligonucleotide Microchip. J. Clin. Microbiol. 40: 753-757 [Abstract] [Full Text]  
  • Loparev, V. N., Massung, R. F., Esposito, J. J., Meyer, H. (2001). Detection and Differentiation of Old World Orthopoxviruses: Restriction Fragment Length Polymorphism of the crmB Gene Region. J. Clin. Microbiol. 39: 94-100 [Abstract] [Full Text]  
  • Henderson, D. A., Inglesby, T. V., Bartlett, J. G., Ascher, M. S., Eitzen, E., Jahrling, P. B., Hauer, J., Layton, M., McDade, J., Osterholm, M. T., O'Toole, T., Parker, G., Perl, T., Russell, P. K., Tonat, K., for the Working Group on Civilian Biodefense, (1999). Smallpox as a Biological Weapon: Medical and Public Health Management. JAMA 281: 2127-2137 [Abstract] [Full Text]  
  • Breman, J. G., Henderson, D.A. (2002). Diagnosis and Management of Smallpox. NEJM 346: 1300-1308 [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»