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 Pehler-Harrington, K.
Right arrow Articles by Henrickson, K. J.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Pehler-Harrington, K.
Right arrow Articles by Henrickson, K. J.

 Previous Article  |  Next Article 

Journal of Clinical Microbiology, September 2004, p. 4072-4076, Vol. 42, No. 9
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.9.4072-4076.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Rapid Detection and Identification of Human Adenovirus Species by Adenoplex, a Multiplex PCR-Enzyme Hybridization Assay

Karen Pehler-Harrington,1 Marilyn Khanna,1 Chris R. Waters,1 and Kelly J. Henrickson1,2*

Prodesse, Inc., Waukesha,1 Department of Pediatrics, Medical College of Wisconsin, Milwaukee, Wisconsin2

Received 19 November 2003/ Returned for modification 29 December 2003/ Accepted 8 May 2004

Human adenoviruses (AdV) have been implicated in a wide variety of diseases and are ubiquitous in populations worldwide. These agents are of concern particularly in immunocompromised patients, children, and military recruits, resulting in severe disease or death. Clinical diagnosis of AdV is usually achieved through routine viral cell culture, which can take weeks for results. Immunofluorescence and enzyme-linked immunosorbent assay-based techniques are more timely but lack sensitivity. The ability to distinguish between the six different AdV species (A to F) is diagnostically relevant, as infections with specific AdV species are often associated with unique clinical outcomes and epidemiological features. Therefore, we developed a multiplex PCR-enzyme hybridization assay, the Adenoplex, using primers to the fiber gene that can simultaneously detect all six AdV species A through F in a single test. The limit of detection (LOD) based on the viral 50% tissue culture infective dose/ml for AdV A, B, C, D, E, and F was 10–2, 10–1, 10–1, 10–2, 10–1, and 10–2, respectively. Similarly, the LOD for the six DNA controls ranged from 102 to 103 copies/ml. Twelve common respiratory pathogens were tested with the Adenoplex, and no cross-reactivity was observed. We also validated our assay using clinical specimens spiked with different concentrations of AdV strains of each species type and tested by multiplex PCR and culture. The results demonstrated an overall sensitivity and specificity of Adenoplex of 100%. This assay can be completed in as few as 5 h and provides a rapid, specific, and sensitive method to detect and subtype AdV species A through F.

* Corresponding author. Mailing address: Department of Pediatrics, MACC Fund Research Center, Medical College of Wisconsin, 8701 Watertown Plank Rd., Milwaukee, WI 53226. Phone: (414) 456-4122. Fax: (414) 456-6539. E-mail: kellyj{at}mcw.edu.

Journal of Clinical Microbiology, September 2004, p. 4072-4076, Vol. 42, No. 9
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.9.4072-4076.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Russell, W. C. (2009). Adenoviruses: update on structure and function. J. Gen. Virol. 90: 1-20 [Abstract] [Full Text]  
  • Damen, M., Minnaar, R., Glasius, P., van der Ham, A., Koen, G., Wertheim, P., Beld, M. (2008). Real-Time PCR with an Internal Control for Detection of All Known Human Adenovirus Serotypes. J. Clin. Microbiol. 46: 3997-4003 [Abstract] [Full Text]  
  • Mahony, J. B. (2008). Detection of Respiratory Viruses by Molecular Methods. Clin. Microbiol. Rev. 21: 716-747 [Abstract] [Full Text]  
  • Kumar, S., Wang, L., Fan, J., Kraft, A., Bose, M. E., Tiwari, S., Van Dyke, M., Haigis, R., Luo, T., Ghosh, M., Tang, H., Haghnia, M., Mather, E. L., Weisburg, W. G., Henrickson, K. J. (2008). Detection of 11 Common Viral and Bacterial Pathogens Causing Community-Acquired Pneumonia or Sepsis in Asymptomatic Patients by Using a Multiplex Reverse Transcription-PCR Assay with Manual (Enzyme Hybridization) or Automated (Electronic Microarray) Detection. J. Clin. Microbiol. 46: 3063-3072 [Abstract] [Full Text]  
  • Reijans, M., Dingemans, G., Klaassen, C. H., Meis, J. F., Keijdener, J., Mulders, B., Eadie, K., van Leeuwen, W., van Belkum, A., Horrevorts, A. M., Simons, G. (2008). RespiFinder: a New Multiparameter Test To Differentially Identify Fifteen Respiratory Viruses. J. Clin. Microbiol. 46: 1232-1240 [Abstract] [Full Text]  
  • Letant, S. E., Ortiz, J. I., Bentley Tammero, L. F., Birch, J. M., Derlet, R. W., Cohen, S., Manning, D., McBride, M. T. (2007). Multiplexed Reverse Transcriptase PCR Assay for Identification of Viral Respiratory Pathogens at the Point of Care. J. Clin. Microbiol. 45: 3498-3505 [Abstract] [Full Text]  
  • Lee, W.-M., Grindle, K., Pappas, T., Marshall, D. J., Moser, M. J., Beaty, E. L., Shult, P. A., Prudent, J. R., Gern, J. E. (2007). High-Throughput, Sensitive, and Accurate Multiplex PCR-Microsphere Flow Cytometry System for Large-Scale Comprehensive Detection of Respiratory Viruses. J. Clin. Microbiol. 45: 2626-2634 [Abstract] [Full Text]  
  • Madisch, I., Harste, G., Pommer, H., Heim, A. (2005). Phylogenetic Analysis of the Main Neutralization and Hemagglutination Determinants of All Human Adenovirus Prototypes as a Basis for Molecular Classification and Taxonomy. J. Virol. 79: 15265-15276 [Abstract] [Full Text]  
  • Lee, J. A, Kim, N. H., Kim, S. J., Choi, E. H., Lee, H. J. (2005). Rapid Identification of Human Adenovirus Types 3 and 7 from Respiratory Specimens via Multiplex Type-Specific PCR. J. Clin. Microbiol. 43: 5509-5514 [Abstract] [Full Text]  
  • Garcia, E. P., Dowding, L. A., Stanton, L. W., Slepnev, V. I. (2005). Scalable Transcriptional Analysis Routine--Multiplexed Quantitative Real-Time Polymerase Chain Reaction Platform for Gene Expression Analysis and Molecular Diagnostics. J. Mol. Diagn. 7: 444-454 [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»