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 Osiowy, C.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Osiowy, C.

 Previous Article  |  Next Article 

Journal of Clinical Microbiology, November 1998, p. 3149-3154, Vol. 36, No. 11
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Direct Detection of Respiratory Syncytial Virus, Parainfluenza Virus, and Adenovirus in Clinical Respiratory Specimens by a Multiplex Reverse Transcription-PCR Assay

Carla Osiowy*

Influenza and Respiratory Viruses, Bureau of Microbiology, Laboratory Center for Disease Control, Federal Laboratories, Winnipeg, Manitoba R3M 3R2, Canada

Received 28 April 1998/Returned for modification 26 June 1998/Accepted 18 August 1998

Diagnosis of respiratory virus infections currently involves detection by isolation or antigen detection, which usually identifies only a single suspected agent. To permit identification of more than one respiratory virus in clinical specimens, a rapid detection method involving a single-step, multiplex reverse transcription-PCR (RT-PCR) assay was developed. The assay included five primer sets that amplified the RNA of respiratory syncytial virus subtypes A and B, parainfluenza virus types 1, 2, and 3, and adenovirus types 1 to 7. Initially the assay was tested on tissue culture-grown virus and was found to be specific for all 12 prototype viruses tested, with no interassay cross amplification or amplification of other respiratory viruses. Assay sensitivity allowed a detection range of 0.2 50% tissue culture infectious dose (TCID50) for adenovirus to 250 TCID50 for parainfluenza virus type 1. The multiplex RT-PCR assay was also able to directly detect viruses in respiratory specimens, with virus being detected in 41 of 112 samples as compared to 34 of 112 samples detected by direct immunofluorescence or antigen detection following specimen culture. This suggests that the multiplex RT-PCR assay can be used as a rapid and sensitive diagnostic method for major respiratory viruses.

* Present address: Bloodborne Pathogens and Hepatitis, Bureau of Microbiology, Laboratory Center for Disease Control, Federal Laboratories, 1015 Arlington St., Winnipeg, Manitoba R3M 3R2, Canada. Phone: (204) 789-6061. Fax: (204) 789-2082. E-mail: carla_osiowy{at}hc-sc.gc.ca.

Journal of Clinical Microbiology, November 1998, p. 3149-3154, Vol. 36, No. 11
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Hasman, H., Pachucki, C. T., Unal, A., Nguyen, D., Devlin, T., Peeples, M. E., Kwilas, S. A. (2009). Aetiology of influenza-like illness in adults includes parainfluenzavirus type 4. J Med Microbiol 58: 408-413 [Abstract] [Full Text]  
  • Mahony, J. B. (2008). Detection of Respiratory Viruses by Molecular Methods. Clin. Microbiol. Rev. 21: 716-747 [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]  
  • Lam, W. Y., Yeung, A. C. M., Tang, J. W., Ip, M., Chan, E. W. C., Hui, M., Chan, P. K. S. (2007). Rapid Multiplex Nested PCR for Detection of Respiratory Viruses. J. Clin. Microbiol. 45: 3631-3640 [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]  
  • Endo, R., Ishiguro, N., Kikuta, H., Teramoto, S., Shirkoohi, R., Ma, X., Ebihara, T., Ishiko, H., Ariga, T. (2007). Seroepidemiology of Human Bocavirus in Hokkaido Prefecture, Japan. J. Clin. Microbiol. 45: 3218-3223 [Abstract] [Full Text]  
  • Mahony, J., Chong, S., Merante, F., Yaghoubian, S., Sinha, T., Lisle, C., Janeczko, R. (2007). Development of a Respiratory Virus Panel Test for Detection of Twenty Human Respiratory Viruses by Use of Multiplex PCR and a Fluid Microbead-Based Assay. J. Clin. Microbiol. 45: 2965-2970 [Abstract] [Full Text]  
  • Li, H., McCormac, M. A., Estes, R. W., Sefers, S. E., Dare, R. K., Chappell, J. D., Erdman, D. D., Wright, P. F., Tang, Y.-W. (2007). Simultaneous Detection and High-Throughput Identification of a Panel of RNA Viruses Causing Respiratory Tract Infections. J. Clin. Microbiol. 45: 2105-2109 [Abstract] [Full Text]  
  • Lin, B., Blaney, K. M., Malanoski, A. P., Ligler, A. G., Schnur, J. M., Metzgar, D., Russell, K. L., Stenger, D. A. (2007). Using a Resequencing Microarray as a Multiple Respiratory Pathogen Detection Assay. J. Clin. Microbiol. 45: 443-452 [Abstract] [Full Text]  
  • Harju, T H, Leinonen, M, Nokso-Koivisto, J, Korhonen, T, Raty, R, He, Q, Hovi, T, Mertsola, J, Bloigu, A, Rytila, P, Saikku, P (2006). Pathogenic bacteria and viruses in induced sputum or pharyngeal secretions of adults with stable asthma. Thorax 61: 579-584 [Abstract] [Full Text]  
  • Ebihara, T., Endo, R., Ma, X., Ishiguro, N., Kikuta, H. (2005). Detection of Human Metapneumovirus Antigens in Nasopharyngeal Secretions by an Immunofluorescent-Antibody Test. J. Clin. Microbiol. 43: 1138-1141 [Abstract] [Full Text]  
  • Gruteke, P., Glas, A. S., Dierdorp, M., Vreede, W. B., Pilon, J.-W., Bruisten, S. M. (2004). Practical Implementation of a Multiplex PCR for Acute Respiratory Tract Infections in Children. J. Clin. Microbiol. 42: 5596-5603 [Abstract] [Full Text]  
  • Syrmis, M. W., Whiley, D. M., Thomas, M., Mackay, I. M., Williamson, J., Siebert, D. J., Nissen, M. D., Sloots, T. P. (2004). A Sensitive, Specific, and Cost-Effective Multiplex Reverse Transcriptase-PCR Assay for the Detection of Seven Common Respiratory Viruses in Respiratory Samples. J. Mol. Diagn. 6: 125-131 [Abstract] [Full Text]  
  • Boivin, G., Cote, S., Dery, P., De Serres, G., Bergeron, M. G. (2004). Multiplex Real-Time PCR Assay for Detection of Influenza and Human Respiratory Syncytial Viruses. J. Clin. Microbiol. 42: 45-51 [Abstract] [Full Text]  
  • Rocholl, C., Gerber, K., Daly, J., Pavia, A. T., Byington, C. L. (2004). Adenoviral Infections in Children: The Impact of Rapid Diagnosis. Pediatrics 113: e51-56 [Abstract] [Full Text]  
  • Erdman, D. D., Weinberg, G. A., Edwards, K. M., Walker, F. J., Anderson, B. C., Winter, J., Gonzalez, M., Anderson, L. J. (2003). GeneScan Reverse Transcription-PCR Assay for Detection of Six Common Respiratory Viruses in Young Children Hospitalized with Acute Respiratory Illness. J. Clin. Microbiol. 41: 4298-4303 [Abstract] [Full Text]  
  • Cote, S., Abed, Y., Boivin, G. (2003). Comparative Evaluation of Real-Time PCR Assays for Detection of the Human Metapneumovirus. J. Clin. Microbiol. 41: 3631-3635 [Abstract] [Full Text]  
  • Whiley, D. M., Syrmis, M. W., Mackay, I. M., Sloots, T. P. (2002). Detection of Human Respiratory Syncytial Virus in Respiratory Samples by LightCycler Reverse Transcriptase PCR. J. Clin. Microbiol. 40: 4418-4422 [Abstract] [Full Text]  
  • Boivin, G., Osterhaus, A. D., Gaudreau, A., Jackson, H. C., Groen, J., Ward, P. (2002). Role of Picornaviruses in Flu-Like Illnesses of Adults Enrolled in an Oseltamivir Treatment Study Who Had No Evidence of Influenza Virus Infection. J. Clin. Microbiol. 40: 330-334 [Abstract] [Full Text]  
  • Liolios, L., Jenney, A., Spelman, D., Kotsimbos, T., Catton, M., Wesselingh, S. (2001). Comparison of a Multiplex Reverse Transcription-PCR-Enzyme Hybridization Assay with Conventional Viral Culture and Immunofluorescence Techniques for the Detection of Seven Viral Respiratory Pathogens. J. Clin. Microbiol. 39: 2779-2783 [Abstract] [Full Text]  
  • Kehl, S. C., Henrickson, K. J., Hua, W., Fan, J. (2001). Evaluation of the Hexaplex Assay for Detection of Respiratory Viruses in Children. J. Clin. Microbiol. 39: 1696-1701 [Abstract] [Full Text]  
  • AARON, S. D., ANGEL, J. B., LUNAU, M., WRIGHT, K., FEX, C., LE SAUX, N., DALES, R. E. (2001). Granulocyte Inflammatory Markers and Airway Infection during Acute Exacerbation of Chronic Obstructive Pulmonary Disease. Am. J. Respir. Crit. Care Med. 163: 349-355 [Abstract] [Full Text]  
  • Xu, W., McDonough, M. C., Erdman, D. D. (2000). Species-Specific Identification of Human Adenoviruses by a Multiplex PCR Assay. J. Clin. Microbiol. 38: 4114-4120 [Abstract] [Full Text]  
  • Elnifro, E. M., Ashshi, A. M., Cooper, R. J., Klapper, P. E. (2000). Multiplex PCR: Optimization and Application in Diagnostic Virology. Clin. Microbiol. Rev. 13: 559-570 [Abstract] [Full Text]  
  • Louie, M., Louie, L., Simor, A. E. (2000). The role of DNA amplification technology in the diagnosis of infectious diseases. CMAJ 163: 301-309 [Abstract] [Full Text]  
  • Aguilar, J. C., Pérez-Breña, M. P., García, M. L., Cruz, N., Erdman, D. D., Echevarría, J. E. (2000). Detection and Identification of Human Parainfluenza Viruses 1, 2, 3, and 4 in Clinical Samples of Pediatric Patients by Multiplex Reverse Transcription-PCR. J. Clin. Microbiol. 38: 1191-1195 [Abstract] [Full Text]  
  • Sullender, W. M. (2000). Respiratory Syncytial Virus Genetic and Antigenic Diversity. Clin. Microbiol. Rev. 13: 1-15 [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»