Optimisation of the PEG reconcentration procedure for virus detection by cell culture or genomic amplification

Vilagines, P.; Suarez, A.; Sarrette, B.; Vilagines, R.

doi:10.2166/wst.1997.0777

Cited by 13 publications

(7 citation statements)

References 0 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Water samples were kept between 4 and 10°C during transport to the laboratory and stored at −80°C upon arrival. For AIV detection, each water sample was processed for concentration and precipitation of any potential viral material using PEG6000 as previously described [21] . Briefly, 2 litres of water were mixed with 200 g of PEG (Polyethylene glycol, 6000 BioUltra, Sigma-Adrich, Madrid, Spain) and adjusted to 0.3 M NaCl concentration.…”

Section: Methodsmentioning

confidence: 99%

Ecological Factors Driving Avian Influenza Virus Dynamics in Spanish Wetland Ecosystems

et al. 2012

View full text Add to dashboard Cite

Studies exploring the ecological interactions between avian influenza viruses (AIV), natural hosts and the environment are scarce. Most work has focused on viral survival and transmission under laboratory conditions and through mathematical modelling. However, more integrated studies performed under field conditions are required to validate these results. In this study, we combined information on bird community, environmental factors and viral epidemiology to assess the contribution of biotic and abiotic factors in the occurrence of low pathogenic AIV in Spanish wetlands. For that purpose, seven locations in five different wetlands were studied during two years (2007–2009), including seven sampling visits by location. In each survey, fresh faeces (n = 4578) of wild birds and water samples were collected for viral detection. Also, the vegetation structure, water physical properties of wetlands, climatic conditions and wild bird community composition were determined. An overall AIV prevalence of 1.7%±0.4 was detected in faecal samples with important fluctuations among seasons and locations. Twenty-six AIV were isolated from the 78 RRT-PCR positive samples and eight different haemagglutinines and five neuraminidases were identified, being the combination H3N8 the most frequent. Variation partitioning procedures identified the combination of space and time variables as the most important pure factor – independently to other factors – explaining the variation in AIV prevalence (36.8%), followed by meteorological factor (21.5%) and wild bird community composition/vegetation structure (21.1%). These results contribute to the understanding of AIV ecological drivers in Spanish ecosystems and provide useful guidelines for AIV risk assessment identifying potential hotspots of AIV activity.

show abstract

Section: Methodsmentioning

confidence: 99%

Ecological Factors Driving Avian Influenza Virus Dynamics in Spanish Wetland Ecosystems

et al. 2012

View full text Add to dashboard Cite

show abstract

“…The pH of the eluate was adjusted to neutral with 1 M HCl. In the secondary concentration step, the 100 mL eluate was concentrated to a final volume of 20 mL in sterile phosphate buffered saline (pH 7.4, Sigma-Aldrich Co., USA) by polyethylene glycol (PEG)/sodium chloride (NaCl) precipitation, as described by Minor [28] and Vilagines et al [29]. The recovered concentrate was stored at −20 • C until further processing.…”

Section: Pathogenic Viral Detectionmentioning

confidence: 99%

Human Enteric Pathogens in Eight Rivers Used as Rural Household Drinking Water Sources in the Northern Region of South Africa

Potgieter

Karambwe

Mudau

et al. 2020

IJERPH

View full text Add to dashboard Cite

People living in rural areas still rely on the use of environmental water that is contaminated by human and animal activities. This study assessed the occurrence of human enteric pathogens in rivers that are used by rural communities Vhembe District of South Africa as a source of drinking water covering two seasons (winter and summer) over a one-year period. Water quality was assessed using physico characteristics and indicator organisms (total coliforms, E. coli, Clostridium perfringens). Pathogens tested included bacteria (Pathogenic E. coli, Salmonella-, Shigella- and Vibrio spp.), protozoa (Cryptosporidium- and Giardia spp.), and enteric viruses (Rota-, Noro-, Entero-, and Adenoviruses) while using published molecular protocols. The results showed that the indicator bacteria counts exceeded South African drinking water quality guideline limits and pathogenic E. coli was detected in the samples. No Shigella spp. were isolated, while Vibrio spp. and Salmonella spp. were present; parasites were detected in four rivers and Enteric viruses were predominantly detected in the winter season. The results indicated the poor condition of water and the potential health risks to consumers highlighting the need for implementing river catchment management strategies for continued sustainability in these rivers.

show abstract

“…Unlike cell culture, PCR enables simultaneous detection of a number of viruses, and also has the advantage of being able to distinguish between virus genotypes. However, PCR may not distinguish between infectious and non‐infectious virus particles, so may not provide a useful measure of public health risk ( Alexander and Morris 1991; Vilagrines et al . 1997 ).…”

Section: Introductionmentioning

confidence: 99%

Influence of environmental factors on virus detection by RT-PCR and cell culture

et al. 2000

View full text Add to dashboard Cite

The effects of clay, humic acid, u.v. light and shellfish tissue residues on the detection of poliovirus type 2 from environmental samples by culture and RT‐PCR were investigated. RT‐PCR showed 10–100 times greater sensitivity for PV2 detection in the absence of sample contaminants than did culture by plaque assay in BGM cell monolayers. Bentonite clay (100–1000 mg l−1) and shellfish tissue residues reduced virus detection by plaque assay, but the effect of bentonite was mitigated by simple elution procedures. Bentonite clay, humic acid (5–150 mg l−1) and mussel tissue reduced virus detection by RT‐PCR by between 1 and 8 logs, although this was mitigated in part by elution and Sephadex filtration of extracts. Sephadex filtration of samples reduced culturable PV2 by 32–50%. Exposure of PV2 in water to u.v. light reduced culturability of PV2 but not detection by RT‐PCR. This study demonstrates that virus detection in environmental samples is strongly influenced by naturally occurring substances and disinfection approaches. The accuracy of results of viral analyses of this nature should be carefully scrutinized with respect to sample constituents.

show abstract

Optimisation of the PEG reconcentration procedure for virus detection by cell culture or genomic amplification

Cited by 13 publications

References 0 publications

Ecological Factors Driving Avian Influenza Virus Dynamics in Spanish Wetland Ecosystems

Ecological Factors Driving Avian Influenza Virus Dynamics in Spanish Wetland Ecosystems

Human Enteric Pathogens in Eight Rivers Used as Rural Household Drinking Water Sources in the Northern Region of South Africa

Influence of environmental factors on virus detection by RT-PCR and cell culture

Contact Info

Product

Resources

About