Detection and differentiation of old world orthopoxviruses : Restriction fragment length polymorphism of the crmB gene region

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...

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Bibliographic Details
Published in:Journal of clinical microbiology 2001, Vol.39 (1), p.94-100
Main Authors: LOPAREV, Vladimir N, MASSUNG, Robert F, ESPOSITO, Joseph J, MEYER, Hermann
Format: Article
Language:English
Subjects:
Biological and medical sciences
Camelpox virus
Cowpox virus
crmB gene
Fundamental and applied biological sciences. Psychology
Humans
Microbiology
Monkeypox virus
Orthopoxvirus - classification
Orthopoxvirus - genetics
Orthopoxvirus - isolation & purification
Polymerase Chain Reaction - methods
Polymorphism, Restriction Fragment Length
Poxviridae Infections - diagnosis
Poxviridae Infections - virology
Receptors, Tumor Necrosis Factor - genetics
Techniques used in virology
Vaccinia virus
Variola virus
Viral Proteins - genetics
Virology
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Summary: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.
ISSN:0095-1137
1098-660X
DOI:10.1128/jcm.39.1.94-100.2001