Improved Detection of Norovirus and Hepatitis A Virus in Surface Water by Applying Pre-PCR Processing

2017 ◽  
Vol 9 (4) ◽  
pp. 395-405 ◽  
Author(s):  
Emmy Borgmästars ◽  
Mehrdad Mousavi Jazi ◽  
Sofia Persson ◽  
Linda Jansson ◽  
Peter Rådström ◽  
...  
Keyword(s):  
Surface Water ◽  
Hepatitis A ◽  
Hepatitis A Virus

Detection of Hepatitis a Virus and other Enteroviruses in Wastewater and Surface Water Samples by Gene Probe Assay

1991 ◽  
Vol 24 (2) ◽  
pp. 267-272 ◽  
Author(s):  
S. Dubrou ◽  
H. Kopecka ◽  
J. M. Lopez Pila ◽  
J. Maréchal ◽  
J. Prévot
Keyword(s):  
Surface Water ◽  
Cell Cultures ◽  
Hepatitis A ◽  
Hepatitis A Virus ◽  
Limit Of Detection ◽  
Blot Hybridization ◽  
Noncoding Region ◽  
Gene Probe ◽  
Dot Blot ◽  
Lower Efficiency

Enteroviruses were specifically detected by dot blot hybridization when using poliovirus type 1 (PV1) derived subgenomic radiolabeled cRNA probes (riboprobes) in environmental water specimens and in the cell cultures in which the viruses were amplificated. The riboprobe corresponding to the 5' noncoding sequence detected the majority of enteroviruses. Hepatitis A virus (HAV) was specifically detected by an HAV cRNA probe corresponding to the 5' noncoding region of its genome. By this test, the limit of detection of coxsackievirus B5 and echovirus 7 seeded in mineral water was 103 to 104 PFU/spot. In cell cultures, positive signals were observed in the lysates of cells infected by one PFU. Higher positive signals were obtained with a short PV1 probe (nt 221-670) corresponding to the 5' noncoding region, which is a well preserved sequence among the enteroviruses, than with PV1 genomic probe. Hybridization allowed a good detection of enteroviral RNAs in wastewater specimens, but with a lower efficiency in surface water. In this case, amplification of viruses in the cell cultures gave significant hybridization results.

Use of RT seminested PCR to assess viral contamination in Caribbean rivers (Martinique)

1995 ◽  
Vol 31 (5-6) ◽  
pp. 391-394 ◽  
Author(s):  
F. Le Guyader ◽  
D. Menard ◽  
M. Pommepuy ◽  
H. Kopecka
Keyword(s):  
Surface Water ◽  
Hepatitis A ◽  
Bacterial Contamination ◽  
Hepatitis A Virus ◽  
Caribbean Island ◽  
Good Tool ◽  
Surface Water Pollution ◽  
Viral Contamination ◽  
Virus Rna ◽  
Seminested Pcr

RT seminested PCR was used to detect enterovirus and hepatitis A virus RNA in polluted surface water in a Caribbean island. Similar results were obtained from samples with or without PEG concentrations: viral RNA being detected in about 60% of river samples. No correlation was found with bacterial contamination, but some inhibitors may have been present. PCR would seem to be a good tool for monitoring surface water pollution.

Surveillance of human viral contamination and physicochemical profiles in a surface water lagoon

2012 ◽  
Vol 66 (12) ◽  
pp. 2682-2687 ◽  
Author(s):  
G. Fongaro ◽  
M. A Nascimento ◽  
A. Viancelli ◽  
D. Tonetta ◽  
M. M. Petrucio ◽  
...  
Keyword(s):  
Surface Water ◽  
Hepatitis A ◽  
Hepatitis A Virus ◽  
Quantitative Polymerase Chain Reaction ◽  
Human Adenovirus ◽  
Human Consumption ◽  
Viral Contamination ◽  
Human Waste ◽  
Viral Particles ◽  
Polymerase Chain

The present study evaluated the contamination of a surface water lagoon (Peri Lagoon) in Florianópolis, Santa Catarina, Brazil, by human adenovirus (HAdV), polyomavirus JC (JCPyV), hepatitis A virus (HAV) and rotavirus species A (RVA). Efforts were driven to determine the correlation between viral presence and the physicochemical parameters of the lagoon and measure the distribution of these viruses throughout the year (June 2010 to May 2011). A total of 48 samples were collected, concentrated and analyzed by qPCR (quantitative polymerase chain reaction). Approximately 96% of the samples were positive for HAdV (46/48), 65% were positive for RVA (31/48), 21% were positive for JCPyV (10/48) and 12% were positive for HAV (6/48). The presence of JCPyV was positively correlated with that of NO2−N, and also there was a positive correlation between the presence of each one of the viruses (HAdV, HAV and RVA) in winter. Samples from water dedicated for human consumption and recreation tested positive for HAdV by qPCR. These samples were also subjected to viral integrity and viability assays: 83% (10/12) contained intact viral particles and 66% (8/12) contained infectious particles. Our results demonstrate the release of human waste into water sources, justifying the urgent need to add viral parameters to water quality surveillance.

scholarly journals Quantitative RT-PCR Detection of Human Noroviruses and Hepatitis A Virus in Surface Water and Fresh Produce

2021 ◽  
Author(s):  
Mohamed N.F. Shaheen ◽  
Elmahdy M. Elmahdy ◽  
Lamiaa H. I. Mahmoud ◽  
Ibtisam A. Hammad ◽  
Elham R.S. Soliman
Keyword(s):  
Surface Water ◽  
Irrigation Water ◽  
Water Samples ◽  
Hepatitis A ◽  
Hepatitis A Virus ◽  
Fresh Produce ◽  
Surface Irrigation ◽  
Human Noroviruses ◽  
Human Enteric Viruses ◽  
Mean Concentration

Abstract Vegetables can be considered as a vehicle for the transmission of human enteric viruses such as noroviruses (genogroups I and II) and hepatitis A virus (HAV) when irrigated with contaminated irrigation or when prepared by infected food handlers. In the current study, we investigated the presence of hepatitis A virus (HAV) and human noroviruses (genogroups I and II) in fresh produce and surface water used in cultivation of this produce using real-time PCR. Samples were collected from six different points in the Mansoura and Giza regions, Egypt. Our analysis showed that at least one virus was found in 41.6% (30/72) of surface water samples and 27% (13/48) of fresh produce samples. HAV (23/72) with a mean viral concentration = 4 × 106 genome copies/litter (GC/L) was the most frequently identified virus in surface irrigation water samples, followed by human norovirus genogroup II (HNoV GII) (15/72, with a mean concentration = 1.2 × 106 GC/L, and human noroviruses genogroup I (HNoV GI) (12/72, with a mean concentration = 1.4 × 104 GC/L). Additionally, HAV (10/48) with a mean concentration = 5.2 × 105 genome copies/gram (GC/g) was also the most frequently detected virus in the fresh produce samples, followed by HNoV GII (8/48, with a mean concentration = 1.7 × 104 GC/g), meanwhile HNoV GI (6/48) was less detected virus with a mean concentration = 3 × 103 GC/g. This work suggests a wide prevalence of human enteric viruses in surface irrigation waters and fresh produce, which is of concern when the fresh produce is eaten raw. Thus, Additional monitoring for viral pathogens in irrigation water and food is needed to increase produce safely.

scholarly journals Molecular characterisation of hepatitis A virus strains from water sources in South Africa

2013 ◽  
Vol 69 (5) ◽  
pp. 923-933 ◽  
Author(s):  
R. Saïd ◽  
M. Wolfaardt ◽  
M. B. Taylor
Keyword(s):  
Surface Water ◽  
South African ◽  
Water Samples ◽  
Hepatitis A ◽  
Hepatitis A Virus ◽  
Water Sources ◽  
Nucleotide Sequence Analysis ◽  
Acid Extraction ◽  
Elution Method ◽  
Surface Water Samples

Hepatitis A virus (HAV) strains found in selected South African (SA) surface waters were characterised to establish what HAV types are circulating in the environment, thus reflecting circulation in the surrounding communities. Surface water samples used for irrigation or domestic purposes, and water samples from the outflow of wastewater plants were collected from six provinces. Viruses were recovered from the samples using a glass wool adsorption-elution method and then further concentrated using polyethylene glycol/sodium chloride precipitation. After automated nucleic acid extraction, samples were analysed for HAV by real-time reverse-transcriptase polymerase chain reaction. HAV strains were genotyped by nucleotide sequence analysis of the capsid gene VP1 and the VP1/P2B junction. HAVs were detected in 76% (16/21) of the surface water samples and in 37% (19/51) of the samples from the wastewater plants. Strains were characterised from 32 of the 35 samples and classified within genotype IB. The presence of genotype IB in the water sources confirms human faecal contamination. Hence, these faecally-contaminated water sources may be a potential transmission route of HAV infection and a potential source of contamination of irrigated fresh produce in SA.

scholarly journals Hepatitis A virus in surface water in South Africa: what are the risks?

2007 ◽  
Vol 5 (2) ◽  
pp. 229-240 ◽  
Author(s):  
J. M. E. Venter ◽  
J. van Heerden ◽  
J. C. Vivier ◽  
W. O. K. Grabow ◽  
M. B. Taylor
Keyword(s):  
South Africa ◽  
Risk Assessment ◽  
Surface Water ◽  
River Water ◽  
Potential Risk ◽  
Hepatitis A ◽  
Hepatitis A Virus ◽  
Recreational Water ◽  
Risk Of Infection ◽  
Us Epa

The aim of this study was to assess the potential risk of infection constituted by HAV to persons using surface dam and river water for domestic and recreational purposes. It estimates the potential risk using a deterministic exponential risk assessment model with mean values and conservative assumptions. Hepatitis A virus was detected in 17.5% of river and 14.9% of dam water samples tested. The number of indicator organisms in these sources exceeded drinking and recreational water quality guidelines set by the United States Environmental Protection Agency (US EPA), indicating possible health risks to recreational water users. Based on the available data and taking all the assumptions into consideration, the probability of infection (Pinf) to the higher socio-economic population using the river water for recreational purposes was 1.1 × 10−3 per day and 3.3 × 10−1 per annum if 100 ml was ingested per day. For recreation in the dam water the Pinf value was 1.2 × 10−4 per day and 4.2 × 10−2 per annum. For the lower socio-economic population, risk values for drinking purposes (2 L day−1) were ten-fold greater. These surface waters therefore did not conform to the US EPA guidelines of 1 infection per 10,000 consumers per year for drinking water or eight gastrointestinal illnesses per 1,000 bathers per day in environmental waters used for recreational purposes. This is the first risk assessment study addressing the risk of infection by HAV in surface water to different socio-economic populations in South Africa.

Application of solid-phase immune electron microscopy to study physical and stability characteristics of enterically transmitted non-a, non-b hepatitis virus

1988 ◽  
Vol 46 ◽  
pp. 80-81
Author(s):  
Charles D. Humphrey ◽  
E. H. Cook ◽  
Karen A. McCaustland ◽  
Daniel W. Bradley
Keyword(s):  
Electron Microscopy ◽  
Hepatitis A ◽  
Hepatitis A Virus ◽  
Solid Phase ◽  
Etiologic Agent ◽  
World Health ◽  
Health Concern ◽  
Morphological Identification ◽  
Immune Electron Microscopy

Enterically transmitted non-A, non-B hepatitis (ET-NANBH) is a type of hepatitis which is increasingly becoming a significant world health concern. As with hepatitis A virus (HAV), spread is by the fecal-oral mode of transmission. Until recently, the etiologic agent had not been isolated and identified. We have succeeded in the isolation and preliminary characterization of this virus and demonstrating that this agent can cause hepatic disease and seroconversion in experimental primates. Our characterization of this virus was facilitated by immune (IEM) and solid phase immune electron microscopic (SPIEM) methodologies.Many immune electron microscopy methodologies have been used for morphological identification and characterization of viruses. We have previously reported a highly effective solid phase immune electron microscopy procedure which facilitated identification of hepatitis A virus (HAV) in crude cell culture extracts. More recently we have reported utilization of the method for identification of an etiologic agent responsible for (ET-NANBH).

Hepatitis A Antigen in Liver, Bile and Stool

1975 ◽  
Vol 33 ◽  
pp. 340-341
Author(s):  
D.R. Jackson ◽  
J.H. Hoofnagle ◽  
A.N. Schulman ◽  
J.L. Dienstag ◽  
R.H. Purcell ◽  
...  
Keyword(s):  
Hepatitis A ◽  
Liver Enzymes ◽  
Hepatitis A Virus ◽  
Type A ◽  
Initial Study ◽  
Virus Like Particle ◽  
Elevated Liver Enzymes ◽  
Ag Particles ◽  
Ag Particle ◽  
Human Stool

Using immune electron microscopy Feinstone et. al. demonstrated the presence of a 27 nm virus-like particle in acute-phase stools of patients with viral hepatitis, type A, These hepatitis A antigen (HA Ag) particles were aggregated by convalescent serum from patients with type A hepatitis but not by pre-infection serum. Subsequently Dienstag et. al. and Maynard et. al. produced acute hepatitis in chimpanzees by inoculation with human stool containing HA Ag. During the early acute disease, virus like particles antigenically, morphologically and biophysically identical to the human HA Ag particle were found in chimpanzee stool. Recently Hilleman et. al. have described similar particles in liver and serum of marmosets infected with hepatitis A virus (HAV). We have investigated liver, bile and stool from chimpanzees and marmosets experimentally infected with HAV. In an initial study, a chimpanzee (no.785) inoculated with HA Ag-containing stool developed elevated liver enzymes 21 days after exposure.

Identification of hepatitis a virus in cell cultures by solid phase immune electron microscopy

1986 ◽  
Vol 44 ◽  
pp. 238-239
Author(s):  
C.D. Humphrey ◽  
T.L. Cromeans ◽  
E.H. Cook ◽  
D.W. Bradley
Keyword(s):  
Electron Microscopy ◽  
Liquid Phase ◽  
Cell Cultures ◽  
Rapid Detection ◽  
Hepatitis A ◽  
Hepatitis A Virus ◽  
Solid Phase ◽  
Immune Electron Microscopy ◽  
Detection And Identification

There is a variety of methods available for the rapid detection and identification of viruses by electron microscopy as described in several reviews. The predominant techniques are classified as direct electron microscopy (DEM), immune electron microscopy (IEM), liquid phase immune electron microscopy (LPIEM) and solid phase immune electron microscopy (SPIEM). Each technique has inherent strengths and weaknesses. However, in recent years, the most progress for identifying viruses has been realized by the utilization of SPIEM.

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