Duplex RT-PCR for simultaneous detection of hepatitis A and hepatitis E virus isolated from drinking water samples

2000 ◽  
Vol 2 (6) ◽  
pp. 587-590 ◽  
Author(s):  
N. Jothikumar ◽  
R. Paulmurugan ◽  
P. Padmanabhan ◽  
R. Balathiripura Sundari ◽  
S. Kamatchiammal ◽  
...  
Keyword(s):  
Drinking Water ◽  
Water Samples ◽  
Hepatitis E Virus ◽  
Hepatitis A ◽  
Simultaneous Detection ◽  
Hepatitis E ◽  
Rt Pcr ◽  
Drinking Water Samples

Study of a hepatitis E virus outbreak involving drinking water and sewage contamination in Shimla, India, 2015–2016

2019 ◽  
Vol 113 (12) ◽  
pp. 789-796 ◽  
Author(s):  
Anuradha S Tripathy ◽  
Meenal Sharma ◽  
Avinash R Deoshatwar ◽  
Prasad Babar ◽  
Rakesh Bharadwaj ◽  
...  
Keyword(s):  
Phylogenetic Analysis ◽  
Water Samples ◽  
Hepatitis E Virus ◽  
Hepatitis A ◽  
Hepatitis A Virus ◽  
Hepatitis E ◽  
Rt Pcr ◽  
Oral Transmission ◽  
Sewage Contamination ◽  
Current Outbreak

Abstract Background Hepatitis E, caused by hepatitis E virus (HEV), accounts for 50% of acute hepatitis cases in India. We report an outbreak of hepatitis E in Shimla, India, in 2015–2016. Methods ICMR-National Institute of Virology (NIV), Pune, received two batches of water samples from Shimla in January 2016 to test for the presence of enterically transmitted hepatitis viruses. Subsequently, 57 icterus patients were tested for various markers of hepatotropic viruses, i.e. anti-HEV IgM/IgG, anti-hepatitis A virus (anti-HAV) IgM/IgG antibodies and HEV RNA. Water samples were screened for HEV and HAV RNA followed by phylogenetic analysis. Results Overall, 48/57 patients availing municipal water had evidence of HEV infection, detected by serology and RT-PCR. All the water samples tested positive for HEV and HAV RNA, while the patients were negative for anti-HAV IgM antibody, indicating no recent HAV infection. Phylogenetic analysis confirmed the aetiological agent of the current outbreak to be HEV genotype 1. Conclusions Serology and RT-PCR confirmed HEV as the aetiology of the outbreak. The absence of new cases of hepatitis A, despite the presence of HAV in the water supply, could be due to previously acquired immunity. Sewage contamination of water leading to faecal-oral transmission of HEV still remains a concern, thus emphasising the need for a vaccination/control strategy.

scholarly journals Occurrence of water-borne enteric viruses in two settlements based in Eastern Chad: analysis of hepatitis E virus, hepatitis A virus and human adenovirus in water sources

2011 ◽  
Vol 9 (3) ◽  
pp. 515-524 ◽  
Author(s):  
Laura Guerrero-Latorre ◽  
Anna Carratala ◽  
Jesus Rodriguez-Manzano ◽  
Byron Calgua ◽  
Ayalkibet Hundesa ◽  
...  
Keyword(s):  
Drinking Water ◽  
Water Samples ◽  
Hepatitis E Virus ◽  
Hepatitis A ◽  
Hepatitis A Virus ◽  
Human Adenovirus ◽  
Hepatitis E ◽  
Water Sources ◽  
Human Adenoviruses ◽  
Water Borne

Hepatitis E virus (HEV) is a common cause of water-borne acute hepatitis in areas with poor sanitation. In 2004 an outbreak of HEV infection affected around 2,000 people in Eastern Chad (Dar Sila). This paper describes the decrease in the incidence of acute jaundice syndrome (AJS) from 2004 until 2009 when a mean incidence of 0.48 cases/1,000 people/year was recorded in the region. Outbreaks of AJS were identified in some of the camps in 2007 and 2008. Moreover, water samples from drinking water sources were screened for human adenoviruses considered as viral indicators and for hepatitis A virus and HEV. Screening of faecal samples from donkeys for HEV gave negative results. Some of the samples were also analysed for faecal coliforms showing values before disinfection treatment between 3 and >50 colony forming units per 100 mL. All water samples tested were negative for HEV and HAV; however, the presence of low levels of human adenoviruses in 4 out of 16 samples analysed indicates possible human faecal contamination of groundwater. Consequently, breakdowns in the treatment of drinking water and/or increased excretion of hepatitis viruses, which could be related to the arrival of a new population, could spread future outbreaks through drinking water.

Development of duplex RT-PCR-ELISA for the simultaneous detection of hepatitis A virus and hepatitis E virus

2011 ◽  
Vol 175 (1) ◽  
pp. 137-140 ◽  
Author(s):  
Hongmin Tahk ◽  
Min Hwa Lee ◽  
Kang Bum Lee ◽  
Doo-Sung Cheon ◽  
Changsun Choi
Keyword(s):  
Hepatitis E Virus ◽  
Hepatitis A ◽  
Hepatitis A Virus ◽  
Simultaneous Detection ◽  
Hepatitis E ◽  
Rt Pcr

scholarly journals Efficiency of Traditional Water Treatment Plant and Compact Units in Removing Viruses

2015 ◽  
Vol 3 (3) ◽  
pp. 528-536
Author(s):  
Yehia A. Osman ◽  
Waled M. El-Senousy ◽  
Adel A. El-Morsi ◽  
Mohammed K. Rashed
Keyword(s):  
Drinking Water ◽  
Water Treatment ◽  
Water Samples ◽  
Treatment Plant ◽  
Water Treatment Plant ◽  
Fecal Bacteria ◽  
Rt Pcr ◽  
Drinking Waters ◽  
Pollution Indicator ◽  
Drinking Water Samples

The fecal bacteria have been taken as the gold standard for water industry. However, the spread of viral gastroenteritis due to drinking water have given a momentum to a recent push by microbiologists to consider viruses as important pollution indicator as fecal bacteria. Therefore, we designed a study to evaluate the efficiency of two types of water purification systems: the traditional water treatment plant and two types compact units. Both systems produced drinking waters free of bacteria, chemical contaminants and mostly viruses free.  However, recent advances in molecular biology techniques, such as RT-PCR have detected Rotaviruses in chlorinated drinking waters resulted from all systems. The frequency of Rotaviruses since October 2010 till September 2012 in Shark El-Mansoura WTP in drinking water samples was 12.5% similar to raw water. While the compact unit at Depo Awam (American design) the frequency of Rotavirus was 16.6% in both raw and drinking water samples.  On the other hand the virus frequency in the raw and drinking water sample in El-Danabik unit (Egyptian design) were 12.5% and 4.16% respectively. Signifying failure of the chlorination process in removing viruses completely.  However, detection of Rotavirus genome in the drinking water samples does not means the presence of its infectivity. The infectious ability of the rotaviruses was confirmed by CC-RT-PCR in all positive samples, where viral RNA was not detected in the collected drinking water samples.  In conclusion RT-PCR and CC-RT-PCR techniques high lightened the need to include viruses as mandatory pollution indicator in water treatment plants. Int J Appl Sci Biotechnol, Vol 3(3): 528-536

scholarly journals Assessment of the Presence of Hepatitis E Virus in Surface Water and Drinking Water in Portugal

2020 ◽  
Vol 8 (5) ◽  
pp. 761 ◽  
Author(s):  
Daniel Salvador ◽  
Célia Neto ◽  
Maria João Benoliel ◽  
Maria Filomena Caeiro
Keyword(s):  
Drinking Water ◽  
Surface Water ◽  
Hepatitis E Virus ◽  
Rna Virus ◽  
Hepatitis E ◽  
Human Consumption ◽  
Contaminated Water ◽  
Cytopathic Effects ◽  
Reverse Transcription Quantitative Pcr ◽  
Drinking Water Samples

Hepatitis E virus (HEV) is a non-enveloped single-stranded positive-sense RNA virus, belonging to the Hepeviridae family, resistant to environmental conditions, and transmitted by the consumption of contaminated water. This virus is responsible for both sporadic and epidemic outbreaks, leading to thousands of infections per year in several countries, and is thus considered an emerging disease in Europe and Asia. This study refers to a survey in Portugal during 2019, targeting the detection and eventual quantification of enteric viruses in samples from surface and drinking water. Samples positive for HEV RNA were recurrently found by reverse transcription quantitative PCR (RT-qPCR), in both types of matrix. The infectivity of these samples was evaluated in cultured Vero E6 cells and RNA from putative viruses produced in cultures evidencing cytopathic effects and was subjected to RT-qPCR targeting HEV genomic RNA. Our results evidenced the existence of samples positive either for HEV RNA (77.8% in surface water and 66.7% in drinking water) or for infectious HEV (23.0% in surface water and 27.7% in drinking water). These results highlight the need for effective virological control of water for human consumption and activities.

A sensitive, semi-quantitative direct PCR-RFLP assay for simultaneous detection of five Cryptosporidium species in treated drinking waters and mineral waters

2002 ◽  
Vol 2 (3) ◽  
pp. 9-15 ◽  
Author(s):  
R.A.B. Nichols ◽  
C.A. Paton ◽  
B.M. Campbell ◽  
J. Wastling ◽  
H.V. Smith
Keyword(s):  
Drinking Water ◽  
Water Samples ◽  
Membrane Filtration ◽  
Simultaneous Detection ◽  
Mineral Waters ◽  
Epifluorescence Microscopy ◽  
Direct Pcr ◽  
Species Identity ◽  
Pcr Rflp ◽  
Drinking Water Samples

We describe a semi-quantitative PCR-RFLP method for detecting low densities of Cryptosporidium spp. oocysts present in final drinking water samples and natural mineral waters. UK Standard Operating Protocols were used to concentrate oocysts from drinking water samples. Oocysts were concentrated from mineral waters by membrane filtration. Cryptosporidium oocysts identified by epifluorescence microscopy on slides or filters were subjected to DNA extraction and PCR-RFLP analysis. Oocysts were disrupted by freeze-thawing in lysis buffer. Amplicons were readily detected from 2 to 5 intact oocysts on ethidium bromide stained gels following 1 round of PCR. DNA extracted from C. parvum, C. muris, C. baileyi, human-derived C. meleagridis, and C. felis were used to confirm species identity by PCR-RFLP following simultaneous digestion with DraI and VspI.

scholarly journals Frequency of HEV contamination in sewerage waters in Pakistan

2010 ◽  
Vol 4 (12) ◽  
pp. 842-845 ◽  
Author(s):  
Tahir Ahmad ◽  
Yasir Waheed ◽  
Sadia Tahir ◽  
Sher Zaman Safi ◽  
Kaneez Fatima ◽  
...  
Keyword(s):  
Water Samples ◽  
Hepatitis E Virus ◽  
Rna Extraction ◽  
Hepatitis E ◽  
Twin Cities ◽  
Rt Pcr ◽  
High Detection Rate ◽  
Drinking Water Resources ◽  
Polymerase Chain ◽  
Routine Procedures

Introduction: Enteric viruses, including Hepatitis E virus (HEV), are able to persist under environmental conditions and may cause public health problems by contaminating natural and drinking water resources. Routine procedures for monitoring viruses in water samples have not been established for the water microbiology screening panel. Methodology: Eighty-six raw sewerage samples were collected from the different regions of Islamabad and Rawalpindi, the twin cities of Pakistan. Samples were concentrated for HEV, using a polyethylene glycol-based method followed by viral RNA extraction using a commercial kit-based method. Reverse transcription polymerase chain reaction (RT-PCR) with HEV specific primers was used for the detection of HEV. Results: The present investigation focused on 86 raw sewerage water samples taken from different locations of drainage outlets of Islamabad and Rawalpindi. After careful experimentation, 35 samples were found to be RT-PCR positive.  Nineteen (44.7%) out of 47 samples from Rawalpindi city were HEV positive while 16 (41.02%) out of 39 samples from Islamabad were HEV positive. All positive samples were found in the highly congested areas. Conclusions: The high detection rate of HEV in this study shows that HEV circulates at a relatively high frequency in the sewerage waters in Pakistan. This study is the first report on detection of HEV from sewerage waste water from Pakistan and suggests that HEV might be a potent indicator of viral pollution in environmental specimens.

Temporal variability of disinfection by-products concentration in urban public water system

2013 ◽  
Vol 14 (4) ◽  
pp. 393-398
Keyword(s):  
Drinking Water ◽  
Water Samples ◽  
Temporal Variability ◽  
Water System ◽  
The Other ◽  
Major Constituent ◽  
System P ◽  
Public Water System ◽  
Drinking Water Samples ◽  
By Products

The occurrence of trihalomethanes (THMs) was studied in the drinking water samples from urban water supply network of Karachi city that served more than 18 million people. Drinking water samples were collected from 58 locations in summer (May-August) and winter (November-February) seasons. The major constituent of THMs detected was chloroform in winter (92.34%) and summer (93.07%), while the other THMs determined at lower concentrations. Summer and winter concentrations of total THMs at places exceed the levels regulated by UEPA (80 μg l-1) and WHO (100 μg l-1). GIS linked temporal variability in two seasons showed significantly higher median concentration (2.5%-23.06%) of THMs compared to winter.

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