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Detection of monkeypox virus with real-time PCR assays.
J Clin Virol. 2006 Jul; 36(3):194-203.JC

Abstract

BACKGROUND

Human monkeypox, a zoonotic disease, was first reported outside of Africa during the 2003 US outbreak.

OBJECTIVES

We present two real-time PCR assays critical for laboratory diagnosis of monkeypox during the 2003 US outbreak.

STUDY DESIGN

A TaqMan-based assay (E9L-NVAR) targets the orthopoxvirus DNA polymerase gene and detects Eurasian orthopoxviruses other than Variola. A hybridization assay, utilizing a MGB Eclipsetrade mark (Epoch Biosciences) probe, targets an envelope protein gene (B6R) and specifically detects monkeypox virus (MPXV). Assays were validated using coded orthopoxvirus DNA samples and used to evaluate lesion samples from five confirmed US monkeypox cases.

RESULTS

E9L-NVAR did not detect variola (48 strains), North American orthopoxviruses (2), or DNA derived from non-poxviral rash illnesses. The assay reproducibly identified various concentrations of 13 Eurasian orthopoxvirus strains and was sensitive to 12.5 vaccinia genomes. The B6R assay recognized 15 different MPXV strains, while other orthopoxvirus (9) and bacteria (15) strains did not cross-react. Of the 13 human samples tested from confirmed cases, both assays identified 100% as containing MPXV DNA.

CONCLUSIONS

E9L-NVAR and B6R assays demonstrate 100% specificity for non-variola Eurasian orthopoxvirus and MPXV, respectively. Using two discrete viral gene targets, these assays together provide a reliable and sensitive method for quickly confirming monkeypox infections.

Authors+Show Affiliations

Poxvirus Program, Division of Viral and Rickettsial Diseases, National Centers of Infectious Diseases, Centers for Disease Control and Prevention, Mail Stop G-43, 1600 Clifton Road, NE, Atlanta, GA 30333, USA.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Evaluation Study
Journal Article

Language

eng

PubMed ID

16731033

Citation

Li, Yu, et al. "Detection of Monkeypox Virus With Real-time PCR Assays." Journal of Clinical Virology : the Official Publication of the Pan American Society for Clinical Virology, vol. 36, no. 3, 2006, pp. 194-203.
Li Y, Olson VA, Laue T, et al. Detection of monkeypox virus with real-time PCR assays. J Clin Virol. 2006;36(3):194-203.
Li, Y., Olson, V. A., Laue, T., Laker, M. T., & Damon, I. K. (2006). Detection of monkeypox virus with real-time PCR assays. Journal of Clinical Virology : the Official Publication of the Pan American Society for Clinical Virology, 36(3), 194-203.
Li Y, et al. Detection of Monkeypox Virus With Real-time PCR Assays. J Clin Virol. 2006;36(3):194-203. PubMed PMID: 16731033.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Detection of monkeypox virus with real-time PCR assays. AU - Li,Yu, AU - Olson,Victoria A, AU - Laue,Thomas, AU - Laker,Miriam T, AU - Damon,Inger K, Y1 - 2006/05/30/ PY - 2005/09/07/received PY - 2006/03/13/revised PY - 2006/03/17/accepted PY - 2006/5/30/pubmed PY - 2006/8/17/medline PY - 2006/5/30/entrez SP - 194 EP - 203 JF - Journal of clinical virology : the official publication of the Pan American Society for Clinical Virology JO - J Clin Virol VL - 36 IS - 3 N2 - BACKGROUND: Human monkeypox, a zoonotic disease, was first reported outside of Africa during the 2003 US outbreak. OBJECTIVES: We present two real-time PCR assays critical for laboratory diagnosis of monkeypox during the 2003 US outbreak. STUDY DESIGN: A TaqMan-based assay (E9L-NVAR) targets the orthopoxvirus DNA polymerase gene and detects Eurasian orthopoxviruses other than Variola. A hybridization assay, utilizing a MGB Eclipsetrade mark (Epoch Biosciences) probe, targets an envelope protein gene (B6R) and specifically detects monkeypox virus (MPXV). Assays were validated using coded orthopoxvirus DNA samples and used to evaluate lesion samples from five confirmed US monkeypox cases. RESULTS: E9L-NVAR did not detect variola (48 strains), North American orthopoxviruses (2), or DNA derived from non-poxviral rash illnesses. The assay reproducibly identified various concentrations of 13 Eurasian orthopoxvirus strains and was sensitive to 12.5 vaccinia genomes. The B6R assay recognized 15 different MPXV strains, while other orthopoxvirus (9) and bacteria (15) strains did not cross-react. Of the 13 human samples tested from confirmed cases, both assays identified 100% as containing MPXV DNA. CONCLUSIONS: E9L-NVAR and B6R assays demonstrate 100% specificity for non-variola Eurasian orthopoxvirus and MPXV, respectively. Using two discrete viral gene targets, these assays together provide a reliable and sensitive method for quickly confirming monkeypox infections. SN - 1386-6532 UR - https://www.unboundmedicine.com/medline/citation/16731033/Detection_of_monkeypox_virus_with_real_time_PCR_assays_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S1386-6532(06)00122-3 DB - PRIME DP - Unbound Medicine ER -