Comparison of a Reporter Assay and Immunomagnetic Separation Real-Time Reverse Transcription-PCR for the Detection of Enteroviruses in Seeded Environmental Water Samples

Hwang, Yu-Chen; Leong, Oymon M.; Chen, Wilfred; Yates, Marylynn V.

doi:10.1128/aem.01758-06

Cited by 32 publications

(13 citation statements)

References 11 publications

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“…Bead-virus complexes can be magnetically separated from the sample and washed to yield concentrated and purified viral particles. Immunomagnetic separation has been used to achieve sensitive detection of EV, RV, HAV and NoV from water, lettuce, green onions, strawberries, shellfish and sewage (Abd el- Galil et al 2005;Arnal et al 1999;Bidawid et al 2000;Casas and Sunen 2002;Gilpatrick et al 2000;Grinde et al 1995;Hwang et al 2007;Jothikumar et al 1998;Kobayashi et al 2004;Lopez-Sabater et al 1997;Monceyron and Grinde 1994;Myrmel et al 2000;Park et al 2008;Shan et al 2005;Sunen and Sobsey 1999). One pitfall of immunomagnetic separation for enteric viruses is that the antibody reagents may be too specific, failing to capture all possible strains of a target virus.…”

Section: Sample Preparationmentioning

confidence: 98%

Analytical Methods for Food and Environmental Viruses

Mattison

Bidawid

2009

Food Environ Virol

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There are many challenges for developing and selecting methods to detect enteric viruses from food and environmental samples. Growth methods are rarely available and the viruses have a low infectious dose, so methods must be very sensitive as well as specific. This review discusses methods for sample preparation, detection and typing, outlining strengths and weaknesses for different protocols. Enteric viruses are very stable in the environment and the development of effective detection methods is an important step towards reducing contamination of foods and the environment.

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Section: Sample Preparationmentioning

confidence: 98%

Analytical Methods for Food and Environmental Viruses

Mattison

Bidawid

2009

Food Environ Virol

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“…Since a "gold standard" for the detection of viruses in environmental samples is not yet defined, the prevalence of viral contamination might be underestimated by our methods (15,16,26). Furthermore, it was shown that the number of samples taken at one site influenced the probability of virus detection, also suggesting more drinking water sources in danger of viral contamination.…”

Section: Discussionmentioning

confidence: 99%

“…In numerous studies, virus concentration from water was achieved by filtration using electropositive filters (1MDS) (12,15,25,26,33). Further methods using ultrafiltration, glass wool filters (7,22), or immunomagnetic separation (16,28) were used to detect small amounts of viruses independently of matrix effects. However, the U.S. Environmental Protection Agency listed adenovirus as one of nine microorganisms on the contamination candidate list for drinking water as a potential indicator virus due to an outstanding resistance to UV disinfection.…”

mentioning

confidence: 99%

Detection of Adenoviruses and Rotaviruses in Drinking Water Sources Used In Rural Areas of Benin, West Africa

Verheyen

Timmen-Wego

Laudien

et al. 2009

Appl Environ Microbiol

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Diseases associated with viruses also found in environmental samples cause major health problems in developing countries. Little is known about the frequency and pattern of viral contamination of drinking water sources in these resource-poor settings. We established a method to analyze 10 liters of water from drinking water sources in a rural area of Benin for the presence of adenoviruses and rotaviruses. Overall, 541 samples from 287 drinking water sources were tested. A total of 12.9% of the sources were positive for adenoviruses and 2.1% of the sources were positive for rotaviruses at least once. Due to the temporary nature of viral contamination in drinking water sources, the probability of virus detection increased with the number of samples taken at one test site over time. No seasonal pattern for viral contaminations was found after samples obtained during the dry and wet seasons were compared. Overall, 3 of 15 surface water samples (20%) and 35 of 247 wells (14.2%) but also 2 of 25 pumps (8%) tested positive for adenoviruses or rotaviruses. The presence of latrines within a radius of 50 m in the vicinity of pumps or wells was identified as being a risk factor for virus detection. In summary, viral contamination was correlated with the presence of latrines in the vicinity of drinking water sources, indicating the importance of appropriate decision support systems in these socioeconomic prospering regions.

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“…An effective concentration of samples is necessary to evaluate virological quality of wastewaters, since matrices are highly contaminated with inhibitory substances that will interfere with viral detection [11]. Numerous methods have been described for concentrating and extracting enteric viruses from sewage [14][15][16]. All of them studied on spiked viral particles in decontaminated samples.…”

Section: Introductionmentioning

confidence: 99%

Comparison of Two Concentration Methods for the Molecular Detection of Enteroviruses in Raw and Treated Sewage

et al. 2015

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Human enteric viruses are a major causative agent of emerging waterborne diseases and constitute a serious public health concern. Environmental contamination occurs through discharge of waste materials from infected persons. Methods for viral detection should be developed to detect low infective dose of enteric viruses in environment. In this study, we aimed at comparing two concentration methods for the detection of naturally occurring enteroviruses in raw and treated sewage. In the first method, polyethylene glycol is used to concentrate viral particles from the collected samples. The second method is based on ultracentrifugation of viral particles at high speed (110,0009g). Genomes of enteroviruses were quantified by the quantitative real-time PCR method in raw and treated sewage samples. PEG-based method yielded higher genomic copies of enteric viruses (with an average of 5.9 log 10 genomic copies/100 mL) when applied to raw sewage samples. While the ultracentrifugation assay in the second method decreases genomic copies number (with an average of 5.4 log 10 genomic copies/100 mL). The recovery differences between the two methods were not significant when applied to clean samples (treated sewage). This could be explained by the presence of inhibitors, which interfere with qRT-PCR, in less quantity comparatively to raw sewage. PEG-based method would be more accurate for samples with highorganic matter load. This report emphasizes the importance of matrices nature on the recovery of enteroviruses from sewage samples. This should be taken into consideration for establishing standardized virological assays to ensure the virological quality control of discharged water in environment.

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Comparison of a Reporter Assay and Immunomagnetic Separation Real-Time Reverse Transcription-PCR for the Detection of Enteroviruses in Seeded Environmental Water Samples

Cited by 32 publications

References 11 publications

Analytical Methods for Food and Environmental Viruses

Analytical Methods for Food and Environmental Viruses

Detection of Adenoviruses and Rotaviruses in Drinking Water Sources Used In Rural Areas of Benin, West Africa

Comparison of Two Concentration Methods for the Molecular Detection of Enteroviruses in Raw and Treated Sewage

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