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 Jean, J.
Right arrow Articles by Fliss, I.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Jean, J.
Right arrow Articles by Fliss, I.
Agricola
Right arrow Articles by Jean, J.
Right arrow Articles by Fliss, I.

 Previous Article  |  Next Article 

Applied and Environmental Microbiology, December 2001, p. 5593-5600, Vol. 67, No. 12
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.12.5593-5600.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Detection of Hepatitis A Virus by the Nucleic Acid Sequence-Based Amplification Technique and Comparison with Reverse Transcription-PCR

Julie Jean,1 Burton Blais,2 André Darveau,3 and Ismaïl Fliss1,*

Centre de Recherche STELA, Département de Sciences des Aliments et de Nutrition,1 Département de Biochimie et Microbiologie,3 Université Laval, Québec G1K 7P4, and Canadian Food Inspection Agency, Laboratory Services Division, Ottawa K1A 0C6,2 Canada

Received 18 May 2001/Accepted 26 September 2001

A nucleic acid sequence-based amplification (NASBA) technique for the detection of hepatitis A virus (HAV) in foods was developed and compared to the traditional reverse transcription (RT)-PCR technique. Oligonucleotide primers targeting the VP1 and VP2 genes encoding the major HAV capsid proteins were used for the amplification of viral RNA in an isothermal process resulting in the accumulation of RNA amplicons. Amplicons were detected by hybridization with a digoxigenin-labeled oligonucleotide probe in a dot blot assay format. Using the NASBA, as little as 0.4 ng of target RNA/ml was detected per comparison to 4 ng/ml for RT-PCR. When crude HAV viral lysate was used, a detection limit of 2 PFU (4 × 102 PFU/ml) was obtained with NASBA, compared to 50 PFU (1 × 104 PFU/ml) obtained with RT-PCR. No interference was encountered in the amplification of HAV RNA in the presence of excess nontarget RNA or DNA. The NASBA system successfully detected HAV recovered from experimentally inoculated samples of waste water, lettuce, and blueberries. Compared to RT-PCR and other amplification techniques, the NASBA system offers several advantages in terms of sensitivity, rapidity, and simplicity. This technique should be readily adaptable for detection of other RNA viruses in both foods and clinical samples.

* Corresponding author. Mailing address: Centre de Recherche STELA, Département de Sciences des Aliments et de Nutrition, Université Laval, Québec (QC) G1K 7P4, Canada. Phone: 418-656-2131, ext. 6825. Fax: 418-656-3353. E-mail: ismail.fliss{at}aln.ulaval.ca.

Applied and Environmental Microbiology, December 2001, p. 5593-5600, Vol. 67, No. 12
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.12.5593-5600.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Lamhoujeb, S., Fliss, I., Ngazoa, S. E., Jean, J. (2008). Evaluation of the Persistence of Infectious Human Noroviruses on Food Surfaces by Using Real-Time Nucleic Acid Sequence-Based Amplification. Appl. Environ. Microbiol. 74: 3349-3355 [Abstract] [Full Text]  
  • Nainan, O. V., Xia, G., Vaughan, G., Margolis, H. S. (2006). Diagnosis of Hepatitis A Virus Infection: a Molecular Approach. Clin. Microbiol. Rev. 19: 63-79 [Abstract] [Full Text]  
  • Abd El Galil, K. H., El Sokkary, M. A., Kheira, S. M., Salazar, A. M., Yates, M. V., Chen, W., Mulchandani, A. (2005). Real-Time Nucleic Acid Sequence-Based Amplification Assay for Detection of Hepatitis A Virus. Appl. Environ. Microbiol. 71: 7113-7116 [Abstract] [Full Text]  
  • Rutjes, S. A., Italiaander, R., van den Berg, H. H. J. L., Lodder, W. J., de Roda Husman, A. M. (2005). Isolation and Detection of Enterovirus RNA from Large-Volume Water Samples by Using the NucliSens miniMAG System and Real-Time Nucleic Acid Sequence-Based Amplification. Appl. Environ. Microbiol. 71: 3734-3740 [Abstract] [Full Text]  
  • Jean, J., D'Souza, D. H., Jaykus, L.-A. (2004). Multiplex Nucleic Acid Sequence-Based Amplification for Simultaneous Detection of Several Enteric Viruses in Model Ready-To-Eat Foods. Appl. Environ. Microbiol. 70: 6603-6610 [Abstract] [Full Text]  
  • Abd El Galil, K. H., El Sokkary, M. A., Kheira, S. M., Salazar, A. M., Yates, M. V., Chen, W., Mulchandani, A. (2004). Combined Immunomagnetic Separation-Molecular Beacon-Reverse Transcription-PCR Assay for Detection of Hepatitis A Virus from Environmental Samples. Appl. Environ. Microbiol. 70: 4371-4374 [Abstract] [Full Text]  
  • Sun, X., Gulyas, M., Hjerpe, A. (2004). Mesothelial Differentiation as Reflected by Differential Gene Expression. Am. J. Respir. Cell Mol. Bio. 30: 510-518 [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»