Enterovirus genomes in wastewater: concentration on glass wool and glass powder and detection by RT-PCR

Gantzer, Christophe; Senouci, Samira; Maul, A.; Lévi, Yves; Schwartzbrod, L.

doi:10.1016/s0166-0934(97)02193-9

Cited by 41 publications

(22 citation statements)

References 24 publications

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“…Glass wool filters have been used by several research teams 3,5,6 to concentrate human enteric viruses from a variety of water sources such as finished drinking water 7 , groundwater 8,9 , surface water 10 , sea water 11 , wastewater 12 , and agricultural runoff 13 . Here we report the filters are also effective in concentrating avian influenza virus as well as the bacterial and protozoan pathogens Salmonella enterica (serovar Typhimurium) and Cryptosporidium parvum, respectively.…”

Section: Discussionmentioning

confidence: 99%

Glass Wool Filters for Concentrating Waterborne Viruses and Agricultural Zoonotic Pathogens

Millen

Gonnering

Berg

et al. 2012

JoVE

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The key first step in evaluating pathogen levels in suspected contaminated water is concentration. Concentration methods tend to be specific for a particular pathogen group, for example US Environmental Protection Agency Method 1623 for Giardia and Cryptosporidium 1 , which means multiple methods are required if the sampling program is targeting more than one pathogen group. Another drawback of current methods is the equipment can be complicated and expensive, for example the VIRADEL method with the 1MDS cartridge filter for concentrating viruses 2 . In this article we describe how to construct glass wool filters for concentrating waterborne pathogens. After filter elution, the concentrate is amenable to a second concentration step, such as centrifugation, followed by pathogen detection and enumeration by cultural or molecular methods. The filters have several advantages. Construction is easy and the filters can be built to any size for meeting specific sampling requirements. The filter parts are inexpensive, making it possible to collect a large number of samples without severely impacting a project budget. Large sample volumes (100s to 1,000s L) can be concentrated depending on the rate of clogging from sample turbidity. The filters are highly portable and with minimal equipment, such as a pump and flow meter, they can be implemented in the field for sampling finished drinking water, surface water, groundwater, and agricultural runoff. Lastly, glass wool filtration is effective for concentrating a variety of pathogen types so only one method is necessary. Here we report on filter effectiveness in concentrating waterborne human enterovirus, Salmonella enterica, Cryptosporidium parvum, and avian influenza virus. Video LinkThe video component of this article can be found at

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Section: Discussionmentioning

confidence: 99%

Glass Wool Filters for Concentrating Waterborne Viruses and Agricultural Zoonotic Pathogens

Millen

Gonnering

Berg

et al. 2012

JoVE

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“…After being mixed, a small volume was immediately sampled for the subsequent virus infectivity assay. The seeded water was pumped by a peristaltic pump from a large plastic jerry can through the tested filter at an average flow rate of 10 or 30 liters/h, as previously used (8,9,28). Two elution procedures were evaluated for elution of viruses from the glass wool and the NanoCeram cartridge filter with 300 ml and 500 ml of an eluting solution, respectively.…”

Section: Methodsmentioning

confidence: 99%

“…Two elution procedures were evaluated for elution of viruses from the glass wool and the NanoCeram cartridge filter with 300 ml and 500 ml of an eluting solution, respectively. The latter consisted of 1.5 or 3% (wt/vol) beef extract (Becton, Dickinson and Company, Le Pont-de-Claix, France) solution (pH 9.5), containing 0 or 0.05 M glycine (Sigma-Aldrich, St. Louis, MO), as previously described (2,9,10,17,19,28). Each elution was performed at an average flow rate of 10 or 30 liters/h, as previously used (8,9,28).…”

Section: Methodsmentioning

confidence: 99%

Development and Validation of a Concentration Method for the Detection of Influenza A Viruses from Large Volumes of Surface Water

Deboosère

Horm

Pinon

et al. 2011

Appl Environ Microbiol

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Contamination of lakes and ponds plays an essential role as a 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 at low levels in water. The aim of this study was to develop and validate a method for the 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 polyethylene glycol (PEG) concentration parameters on the recovery efficiency of the H1N1 strain from 10-liter surface water samples were assessed. An ultimate 1% recovery rate of infectious viruses was achieved with the optimal protocol, corresponding to 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 to 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% with lake water and rainwater, respectively. A theoretical detection threshold of 2.25 ؋ 10 2 TCID 50 (50% tissue culture infectious dose) in the filtered volume was obtained for seeded lake waters by M gene reverse transcriptase PCR (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.

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“…The fibers are inexpensive and require no water conditioning outside of pH adjustment in some circumstances (30,48). Glass wool has been used in virus monitoring studies involving wastewater (10), drinking water (14,41,46), groundwater (6,30,31,43), river water (18,41), and reservoirs (6,43). However, only a handful of studies have attempted to quantify how effective glass wool is for concentrating viruses (7,44,45), and these examined only enteroviruses and rotavirus.…”

mentioning

confidence: 99%

Concentration of Enteroviruses, Adenoviruses, and Noroviruses from Drinking Water by Use of Glass Wool Filters

Lambertini

Spencer

Bertz

et al. 2008

Appl Environ Microbiol

158

153

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Available filtration methods to concentrate waterborne viruses are either too costly for studies requiring large numbers of samples, limited to small sample volumes, or not very portable for routine field applications. Sodocalcic glass wool filtration is a cost-effective and easy-to-use method to retain viruses, but its efficiency and reliability are not adequately understood. This study evaluated glass wool filter performance to concentrate the four viruses on the U.S. Environmental Protection Agency contaminant candidate list, i.e., coxsackievirus, echovirus, norovirus, and adenovirus, as well as poliovirus. Total virus numbers recovered were measured by quantitative reverse transcription-PCR (qRT-PCR); infectious polioviruses were quantified by integrated cell culture (ICC)-qRT-PCR. Recovery efficiencies averaged 70% for poliovirus, 14% for coxsackievirus B5, 19% for echovirus 18, 21% for adenovirus 41, and 29% for norovirus. Virus strain and water matrix affected recovery, with significant interaction between the two variables. Optimal recovery was obtained at pH 6.5. No evidence was found that water volume, filtration rate, and number of viruses seeded influenced recovery. The method was successful in detecting indigenous viruses in municipal wells in Wisconsin. Long-term continuous filtration retained viruses sufficiently for their detection for up to 16 days after seeding for qRT-PCR and up to 30 days for ICC-qRT-PCR. Glass wool filtration is suitable for large-volume samples (1,000 liters) collected at high filtration rates (4 liters min ؊1 ), and its low cost makes it advantageous for studies requiring large numbers of samples.Waterborne viruses are an important cause of disease, being responsible for 14% of outbreaks (9 of 64 cases) and 38% of illnesses (1,153 of 3,008 cases) associated with drinking water in the United States from 1999 to 2002 (21, 49). During the same period, noroviruses were responsible for 6% (8 of 66 cases) of outbreaks and 17% (348 of 2,093 cases) of illnesses associated with recreational water. If waterborne illnesses of unknown etiology during the period 1999-2002 are included in the above statistics, as these are believed to be of viral origin, up to 56% and 28% of illness cases associated with drinking water and recreational water, respectively, may be attributed to viruses.To detect and quantify waterborne viruses from environmental samples, the first step in the protocol usually requires concentration from a large water volume. Several concentration methods have been developed and applied successfully in the past two decades (see reviews by Wyn-Jones and Sellwood [48] and Grabow [13]). These include adsorption onto (and subsequent elution from) electropositive cartridges and membranes (3,25,27,29,33,35), gauze pads and glass powder (2, 9, 34), electronegative membranes, and microporous materials (1,8,12,16,20,27) and concentration by ultrafiltration (15, 17, 36, 37) and ultracentrifugation (26). Adsorption onto electropositive cartridges, for example, the CUNO 1-MDS Viroso...

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Enterovirus genomes in wastewater: concentration on glass wool and glass powder and detection by RT-PCR

Cited by 41 publications

References 24 publications

Glass Wool Filters for Concentrating Waterborne Viruses and Agricultural Zoonotic Pathogens

Glass Wool Filters for Concentrating Waterborne Viruses and Agricultural Zoonotic Pathogens

Development and Validation of a Concentration Method for the Detection of Influenza A Viruses from Large Volumes of Surface Water

Concentration of Enteroviruses, Adenoviruses, and Noroviruses from Drinking Water by Use of Glass Wool Filters

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