tetano
Editor, Senior Moderator
Appl Environ Microbiol. 2011 Apr 15. [Epub ahead of print]
Development and Validation of a Concentration Method for the Detection of Influenza A Viruses from Large Volumes of Surface Water.
Deboosere N, Horm SV, Pinon A, Gachet J, Coldefy C, Buchy P, Vialette M.
Unit? de S?curit? Microbiologique, Institut Pasteur de Lille, 1 rue du Prof. Calmette, BP 245, 59019 Lille, France.
Abstract
Contamination of lakes and ponds plays an essential role as reservoir of avian influenza A virus (AIV) in the environment. A method to concentrate waterborne AIV is a prerequisite for the detection of virus present in water at low levels. The aim of this study was to develop and validate a method for concentration and detection of infectious AIV from large volumes of surface water samples. Two filtration systems, glass wool and electropositive NanoCeram ? filter, were studied. The individual effects of filtration-elution and PEG-concentration parameters on the recovery efficiency of H1N1 strain from 10-liters surface water samples, were assessed. An ultimate 1% recovery rate of infectious viruses was achieved with the optimal protocol, corresponding to a filtration through glass wool, followed by a viral elution step and then a PEG-concentration. This method was validated for the detection of highly pathogenic H5N1 strains from artificially-contaminated larger water volumes, from 10 up to 50 liters, from different sources. The viral recovery efficiencies ranged from 0.01% to 7.89% and from 3.63% to 13.79% from lake and rain waters, respectively. A theoretical detection threshold of 2.25 ? 10(2) TCID(50) in the filtered volume could be obtained for seeded lake waters by M gene RT-PCR. Moreover, the method was used successfully in field studies for the detection of naturally occurring influenza A viruses in lake water in France.
PMID: 21498756 [PubMed - as supplied by publisher]
http://www.ncbi.nlm.nih.gov/pubmed/21498756
Development and Validation of a Concentration Method for the Detection of Influenza A Viruses from Large Volumes of Surface Water.
Deboosere N, Horm SV, Pinon A, Gachet J, Coldefy C, Buchy P, Vialette M.
Unit? de S?curit? Microbiologique, Institut Pasteur de Lille, 1 rue du Prof. Calmette, BP 245, 59019 Lille, France.
Abstract
Contamination of lakes and ponds plays an essential role as reservoir of avian influenza A virus (AIV) in the environment. A method to concentrate waterborne AIV is a prerequisite for the detection of virus present in water at low levels. The aim of this study was to develop and validate a method for concentration and detection of infectious AIV from large volumes of surface water samples. Two filtration systems, glass wool and electropositive NanoCeram ? filter, were studied. The individual effects of filtration-elution and PEG-concentration parameters on the recovery efficiency of H1N1 strain from 10-liters surface water samples, were assessed. An ultimate 1% recovery rate of infectious viruses was achieved with the optimal protocol, corresponding to a filtration through glass wool, followed by a viral elution step and then a PEG-concentration. This method was validated for the detection of highly pathogenic H5N1 strains from artificially-contaminated larger water volumes, from 10 up to 50 liters, from different sources. The viral recovery efficiencies ranged from 0.01% to 7.89% and from 3.63% to 13.79% from lake and rain waters, respectively. A theoretical detection threshold of 2.25 ? 10(2) TCID(50) in the filtered volume could be obtained for seeded lake waters by M gene RT-PCR. Moreover, the method was used successfully in field studies for the detection of naturally occurring influenza A viruses in lake water in France.
PMID: 21498756 [PubMed - as supplied by publisher]
http://www.ncbi.nlm.nih.gov/pubmed/21498756