This Article
Right arrow Full Text
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 Wang, E.
Right arrow Articles by Weaver, S. C.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Wang, E.
Right arrow Articles by Weaver, S. C.

 Previous Article  |  Next Article 

Journal of Clinical Microbiology, November 2006, p. 4000-4008, Vol. 44, No. 11
0095-1137/06/$08.00+0     doi:10.1128/JCM.00175-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Reverse Transcription-PCR-Enzyme-Linked Immunosorbent Assay for Rapid Detection and Differentiation of Alphavirus Infections{triangledown}

Eryu Wang,1,2 Slobodan Paessler,1,2 Patricia V. Aguilar,1,3,{dagger} Anne-Sophie Carrara,1,2 Haolin Ni,1,2 Ivorlyne P. Greene,1,2,{ddagger} and Scott C. Weaver1,2,3*

Center for Biodefense and Emerging Infectious Diseases,1 Department of Pathology,2 Department of Microbiology and Immunology, University of Texas Medical Branch, Galveston, Texas 77555-06093

Received 26 January 2006/ Returned for modification 6 May 2006/ Accepted 22 August 2006

Due to the lack of a rapid, simple, and inexpensive assay for detecting alphavirus infections, we combined a reverse transcription-PCR with an enzyme-linked immunosorbent assay (RT-PCR-ELISA) to identify human pathogenic alphaviruses that are endemic in the New World. By combining the sensitivity of PCR, the detection simplicity of ELISA, and the specificities of DNA probes, this method rapidly detected and differentiated closely related species and subtypes of several medically important alphaviruses. After an amplification using RT-PCR with primers targeting conserved sequences in the nonstructural protein 1 gene, sequence-specific, biotin-labeled probes targeted against Venezuelan, eastern, and western equine encephalitis or Mayaro virus genes were used for the detection of amplicons using ELISA. The assay is simple, fast, and easy to perform in an ordinary diagnostic laboratory or clinical setting. Nucleic acid derived from cell cultures infected with several alphaviruses, clinical specimens, and mosquito pools as well as frozen and paraffin-embedded animal tissues were detected and identified within 6 to 7 h in a sensitive and specific manner.

* Corresponding author. Mailing address: Department of Pathology, University of Texas Medical Branch, Galveston, TX 77555-0609. Phone: (409) 747-0758. Fax: (409) 747-2455. E-mail: sweaver{at}utmb.edu.

{triangledown} Published ahead of print on 6 September 2006.

{dagger} Present address: Department of Microbiology, Mount Sinai School of Medicine, 1 Gustave L. Levy Place, New York, NY 10029.

{ddagger} Present address: W. Harry Feinstone Department of Molecular Microbiology and Immunology, Johns Hopkins Bloomberg School of Public Health, 615 North Wolfe St., Baltimore, MD 21205.

Journal of Clinical Microbiology, November 2006, p. 4000-4008, Vol. 44, No. 11
0095-1137/06/$08.00+0     doi:10.1128/JCM.00175-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Montgomery, S. A., Johnston, R. E. (2007). Nuclear Import and Export of Venezuelan Equine Encephalitis Virus Nonstructural Protein 2. J. Virol. 81: 10268-10279 [Abstract] [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»