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 Traore, O.
Right arrow Articles by Schwartzbrod, L.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Traore, O.
Right arrow Articles by Schwartzbrod, L.
Agricola
Right arrow Articles by Traore, O.
Right arrow Articles by Schwartzbrod, L.

 Previous Article  |  Next Article 

Applied and Environmental Microbiology, August 1998, p. 3118-3122, Vol. 64, No. 8
0099-2240/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Reverse Transcriptase PCR Detection of Astrovirus, Hepatitis A Virus, and Poliovirus in Experimentally Contaminated Mussels: Comparison of Several Extraction and Concentration Methods

Ousmane Traore,1,* Charlotte Arnal,2 Berengere Mignotte,3 Armand Maul,4 Henri Laveran,1 Sylviane Billaudel,2 and Louis Schwartzbrod3

Service d'Hygiène, Faculté de Médecine, 63000 Clermont Fd Cedex,1 Laboratoire de Virologie, Institut de Biologie, Centre Hospitalier Universitaire, 44035 Nantes Cedex 01,2 Laboratoire de Virologie, UMR 7564 CNRS, Faculté de Pharmacie, Université Henri Poincaré Nancy, 54001 Nancy Cedex,3 and Département Statistique et Traitement des Données, Université de Metz, 57045 Metz Cedex,4 France

Received 26 February 1998/Accepted 20 May 1998

Four methods of extraction and three methods of concentration of three enteric viruses from mussels were comparatively evaluated by reverse transcriptase PCR (RT-PCR). Shellfish were experimentally contaminated by immersion in seawater seeded with astrovirus, hepatitis A virus, or poliovirus. Sixty-gram samples of mussel tissues were processed by using borate buffer, glycine solution, saline beef, and saline beef-Freon extraction methods. The viruses were concentrated by precipitation with polyethylene glycol 6000 (PEG 6000) or PEG 8000 or by organic flocculation. RT-PCR was performed with RNA extracts from crude shellfish extracts and concentrates with and without Sephadex LH20 filtration. The glycine solution and borate buffer extraction methods resulted in significantly more RT-PCR-positive samples than the saline beef extraction method. We assessed the efficiency of 20 combinations of extraction and concentration methods. The borate buffer-organic flocculation, borate buffer-PEG 6000, and glycine solution-PEG 6000 combinations gave RT-PCR-positive results for all 27 samples analyzed for the three viruses. Detoxification of the samples by Sephadex LH20 filtration significantly decreased the efficiency of RT-PCR virus detection.

* Corresponding author. Mailing address: Service d'Hygiène, Faculté de Médecine, 28 place Henri Dunant, 63000 Clermont Fd Cedex, France. Phone: (33)4 73 60 80 07. Fax: (33) 4 73 26 54 32. E-mail: Ousmane.Traore{at}u-clermont1.fr.

Applied and Environmental Microbiology, August 1998, p. 3118-3122, Vol. 64, No. 8
0099-2240/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Jones, T. H., Johns, M. W. (2009). Improved Detection of F-Specific RNA Coliphages in Fecal Material by Extraction and Polyethylene Glycol Precipitation. Appl. Environ. Microbiol. 75: 6142-6146 [Abstract] [Full Text]  
  • Myrmel, M., Berg, E. M. M., Rimstad, E., Grinde, B. (2004). Detection of Enteric Viruses in Shellfish from the Norwegian Coast. Appl. Environ. Microbiol. 70: 2678-2684 [Abstract] [Full Text]  
  • Griffin, D. W., Donaldson, K. A., Paul, J. H., Rose, J. B. (2003). Pathogenic Human Viruses in Coastal Waters. Clin. Microbiol. Rev. 16: 129-143 [Abstract] [Full Text]  
  • Shieh, Y.-S. C., Calci, K. R., Baric, R. S. (1999). A Method To Detect Low Levels of Enteric Viruses in Contaminated Oysters. Appl. Environ. Microbiol. 65: 4709-4714 [Abstract] [Full Text]  
  • Arnal, C., Ferre-Aubineau, V., Mignotte, B., Imbert-Marcille, B. M., Billaudel, S. (1999). Quantification of Hepatitis A Virus in Shellfish by Competitive Reverse Transcription-PCR with Coextraction of Standard RNA. Appl. Environ. Microbiol. 65: 322-326 [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»