Use of Internally Controlled Real-Time Genome Amplification for Detection of Variola Virus and Other Orthopoxviruses Infecting Humans

Smallpox, once a devastating disease caused by Variola virus, a member of the Orthopoxvirus genus, was eradicated in 1980. However, the importance of variola virus infections has been stressed widely in the last few years, particularly following recent social events in the world. Today, variola viru...

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Bibliographic Details
Published in:Journal of Clinical Microbiology 2006-12, Vol.44 (12), p.4464-4470
Main Authors: Fedele, C.G, Negredo, A, Molero, F, Sánchez-Seco, M.P, Tenorio, A
Format: Article
Language:English
Subjects:
Animals
Biological and medical sciences
Cell Line
DNA Primers
DNA, Viral - analysis
DNA, Viral - genetics
Fundamental and applied biological sciences. Psychology
Genome, Viral
Humans
Infectious diseases
Medical sciences
Microbiology
Miscellaneous
Orthopoxvirus
Orthopoxvirus - classification
Orthopoxvirus - genetics
Orthopoxvirus - isolation & purification
Polymerase Chain Reaction - methods
Polymerase Chain Reaction - standards
Quality Control
Receptors, Tumor Necrosis Factor - genetics
Reference Standards
Reproducibility of Results
Sensitivity and Specificity
Variola virus
Variola virus - classification
Variola virus - genetics
Variola virus - isolation & purification
Viral Proteins - genetics
Virology
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Summary:Smallpox, once a devastating disease caused by Variola virus, a member of the Orthopoxvirus genus, was eradicated in 1980. However, the importance of variola virus infections has been stressed widely in the last few years, particularly following recent social events in the world. Today, variola virus is considered to be one of the most significant agents with potential use as a biological weapon. In this study we developed an internally controlled real-time PCR assay for rapid detection and simultaneous differentiation of variola virus from other orthopoxviruses. The assay is based on TaqMan 3'-minor groove binder (MGB) chemistry and uses generic primers, designed in highly conserved genomic regions of the crmB gene, and three TaqMan MGB probes designed to identify orthopoxviruses, variola virus, and an internal control. The results obtained suggest that the assay is rapid, sensitive, specific, and suitable for the generic detection of orthopoxviruses and the identification of variola virus and avoids false-negative results in a single reaction tube.
ISSN:0095-1137
1098-660X
1098-5530
DOI:10.1128/JCM.00276-06