scholarly journals Development of Virus Concentration Method Using Negatively Charged Membrane by Alkaline Elution after Acid Rinse.

2002 ◽  
Vol 25 (8) ◽  
pp. 469-475 ◽  
Author(s):  
Hiroyuki KATAYAMA ◽  
Akihiro SHIMASAKI ◽  
Shinichiro OHGAKI
Keyword(s):  
Virus Concentration ◽  
Alkaline Elution ◽  
Concentration Method ◽  
Charged Membrane ◽  
Virus Concentration Method

Determination of naturally occurring noroviruses in coastal seawater by alkaline elution after acid rinse using negatively charged membrane

2004 ◽  
Vol 4 (2) ◽  
pp. 73-77 ◽  
Author(s):  
H. Katayama ◽  
A. Tanaka ◽  
M. Otaki ◽  
S. Ohgaki
Keyword(s):  
Cell Culture ◽  
Fecal Coliforms ◽  
Tokyo Bay ◽  
Virus Concentration ◽  
Alkaline Elution ◽  
Concentration Method ◽  
Naturally Occurring ◽  
Charged Membrane ◽  
Virus Concentration Method

A new procedure for concentrating viruses from seawater using a negatively charged membrane eluting with alkaline solution (NaOH, pH 10.5) after acid rinse (H2SO4, pH 3.0) was applied to determine naturally occurring enteric viruses in seawater in Tokyo bay. The levels of total coliforms and fecal coliforms ranged from 40 to 68000 (cfu/100ml) and from 2 to 32000 (cfu/100ml), respectively. The F-specific phages were not detected from 5 ml of 53 samples out of 61 tested. The levels of indicator microbes were not found to be related to the tide in Tokyo bay. Enteroviruses were not detected by cell culture RTÐPCR, but detected by direct RTÐPCR from approximately 10% of the samples. Noroviruses were found positive from approx. 31% of the winter samples (n=29), whereas only 3% from the summer samples (n=32). These results of direct RTÐPCR were equivalent to determination of Norwalk viruses occuring in 50 ml of seawater. These results suggested that the levels of noroviruses in Tokyo bay were higher in winter than those of enteroviruses The virus concentration method used was proved to be useful for determination of naturally occurring viruses in seawater, especially when applied prior to PCR detection of nonculturable viruses.

An efficient virus concentration method and RT-nested PCR for detection of rotaviruses in environmental water samples

2005 ◽  
Vol 124 (1-2) ◽  
pp. 117-122 ◽  
Author(s):  
Leera Kittigul ◽  
Som Ekchaloemkiet ◽  
Fuangfa Utrarachkij ◽  
Kanokrat Siripanichgon ◽  
Dusit Sujirarat ◽  
...  
Keyword(s):  
Water Samples ◽  
Nested Pcr ◽  
Environmental Water ◽  
Environmental Water Samples ◽  
Virus Concentration ◽  
Concentration Method ◽  
Virus Concentration Method

scholarly journals Presence of human noro- and adenoviruses in river and treated wastewater, a longitudinal study and method comparison

2011 ◽  
Vol 10 (1) ◽  
pp. 87-99 ◽  
Author(s):  
Leena Maunula ◽  
Kirsi Söderberg ◽  
Heli Vahtera ◽  
Veli-Pekka Vuorilehto ◽  
Carl-Henrik von Bonsdorff ◽  
...  
Keyword(s):  
River Water ◽  
Method Comparison ◽  
Average Concentration ◽  
Treated Wastewater ◽  
Virus Concentration ◽  
Reverse Transcription Pcr ◽  
Concentration Method ◽  
Charged Membrane ◽  
Causative Agents ◽  
One Year

Norovirus (NoV) is one of the most common causative agents of waterborne gastroenteritis outbreaks. The main objective of the study was to determine the presence of human NoVs in river water and in treated wastewater (TW) released into the river. During a one-year survey in 2007/2008, NoVs were detected in 30.8% of river samples (20/65), and 40.5% of TW samples (17/45) with a real-time reverse transcription-PCR assay. NoVs were present in the river water in the winter and spring, coinciding with the NoV epidemiological peak in the community and the presence of NoVs in TW. Later in 2009, the concentration method used, pre-filtration with a Waterra filter combined with filtration through a negatively charged membrane, was evaluated against glass wool filtration and freeze-drying for the detection of adenoviruses in river water. The virus amounts measured varied greatly depending on the virus concentration method. The continued monitoring in the spring of 2009 also revealed that the average concentration of noro- and adenoviruses in TW was 2.64 × 103 and 1.29 × 104 pcr units per mL, respectively. No correlation between the presence of viruses and Escherichia coli was found. These results may be useful for risk assessment studies.

scholarly journals Development of a Virus Concentration Method and Its Application to Detection of Enterovirus and Norwalk Virus from Coastal Seawater

2002 ◽  
Vol 68 (3) ◽  
pp. 1033-1039 ◽  
Author(s):  
Hiroyuki Katayama ◽  
Akihiro Shimasaki ◽  
Shinichiro Ohgaki
Keyword(s):  
Pure Water ◽  
Inhibitory Effect ◽  
Tokyo Bay ◽  
Virus Concentration ◽  
Natural Seawater ◽  
Norwalk Virus ◽  
Coastal Seawater ◽  
Reverse Transcription Pcr ◽  
Concentration Method ◽  
Charged Membrane

ABSTRACT We developed a new procedure for concentration of enteric viruses from water using a negatively charged membrane. Rinsing the membrane with 0.5 mM H2SO4 (pH 3.0) in order to elute cations prior to viral elution with 1 mM NaOH (pH 10.5) promoted poliovirus recovery yields from 33 to 95% when applied to pure water and 38 to 89% when applied to natural seawater from Tokyo Bay, Japan, respectively. This method showed average recovery yields of spiked poliovirus of 62% (n = 8) from 1 liter of artificial seawater. This method showed higher recovery yields (>61%) than that of the conventional method using positively charged membrane (6%) when applied to seawater. This method is also free from beef extract elution, which has an inhibitory effect in the subsequent viral genome detection by reverse transcription-PCR. Naturally occurring Norwalk viruses from 2 liters of Tokyo Bay water in winter and infectious enteroviruses from 2 liters of recreational coastal seawater in summer were detected by using this viral concentration method.

scholarly journals Comparison of Concentration Methods for Quantitative Detection of Sewage-Associated Viral Markers in Environmental Waters

2015 ◽  
Vol 81 (6) ◽  
pp. 2042-2049 ◽  
Author(s):  
W. Ahmed ◽  
V. J. Harwood ◽  
P. Gyawali ◽  
J. P. S. Sidhu ◽  
S. Toze
Keyword(s):  
Nucleic Acid ◽  
Tap Water ◽  
Environmental Water ◽  
Quality Analysis ◽  
Source Tracking ◽  
Quantitative Detection ◽  
Virus Concentration ◽  
Acid Extraction ◽  
Environmental Waters ◽  
Concentration Method

ABSTRACTPathogenic human viruses cause over half of gastroenteritis cases associated with recreational water use worldwide. They are relatively difficult to concentrate from environmental waters due to typically low concentrations and their small size. Although rapid enumeration of viruses by quantitative PCR (qPCR) has the potential to greatly improve water quality analysis and risk assessment, the upstream steps of capturing and recovering viruses from environmental water sources along with removing PCR inhibitors from extracted nucleic acids remain formidable barriers to routine use. Here, we compared the efficiency of virus recovery for three rapid methods of concentrating two microbial source tracking (MST) viral markers human adenoviruses (HAdVs) and polyomaviruses (HPyVs) from one liter tap water and river water samples on HA membranes (90 mm in diameter). Samples were spiked with raw sewage, and viral adsorption to membranes was promoted by acidification (method A) or addition of MgCl2(methods B and C). Viral nucleic acid was extracted directly from membranes (method A), or viruses were eluted with NaOH and concentrated by centrifugal ultrafiltration (methods B and C). No inhibition of qPCR was observed for samples processed by method A, but inhibition occurred in river samples processed by B and C. Recovery efficiencies of HAdVs and HPyVs were ∼10-fold greater for method A (31 to 78%) than for methods B and C (2.4 to 12%). Further analysis of membranes from method B revealed that the majority of viruses were not eluted from the membrane, resulting in poor recovery. The modification of the originally published method A to include a larger diameter membrane and a nucleic acid extraction kit that could accommodate the membrane resulted in a rapid virus concentration method with good recovery and lack of inhibitory compounds. The frequently used strategy of viral absorption with added cations (Mg2+) and elution with acid were inefficient and more prone to inhibition, and will result in underestimation of the prevalence and concentrations of HAdVs and HPyVs markers in environmental waters.

scholarly journals Performance evaluation of virus concentration methods for implementing SARS-CoV-2 Wastewater-based epidemiology emphasizing quick data turnaround.

2021 ◽  
Author(s):  
Md Ariful Islam Juel ◽  
Nicholas Stark ◽  
Bridgette Nicolosi ◽  
Jordan Lontai ◽  
Kevin Lambirth ◽  
...  
Keyword(s):  
Membrane Filtration ◽  
Municipal Wastewater ◽  
Turnaround Time ◽  
Optimal Choice ◽  
Virus Concentration ◽  
Emf Method ◽  
Municipal Wastewater Treatment ◽  
Filtration Time ◽  
Concentration Method ◽  
Virus Recovery

Wastewater based epidemiology (WBE) has drawn significant attention as an early warning tool to detect and predict the trajectory of COVID-19 cases in a community, in conjunction with public health data. This means of monitoring for outbreaks has been used at municipal wastewater treatment centers to analyze COVID-19 trends in entire communities, as well as by universities and other community living environments to monitor COVID-19 spread in buildings. Sample concentration is crucial, especially when viral abundance in raw wastewater is below the threshold of detection by RT-qPCR analysis. We evaluated the performance of a rapid ultrafiltration-based virus concentration method using InnovaPrep Cp Select pipette tips and compared this to the established electronegative membrane filtration (EMF) method. We evaluated the sensitivity of SARS-CoV-2 quantification, surrogate virus recovery rate, and sample processing time. Results suggest that the Cp Select concentrator is more efficient at concentrating SARS-CoV-2 from wastewater compared to the EMF method. 30% of samples that tested negative when concentrated with the EMF method produced a positive signal with the Cp Select protocol. Increased recovery of the surrogate virus control using the Cp Select confirms this observation. We optimized the Cp Select protocol by adding AVL lysis buffer and sonication, to increase the recovery of virus. Sonication increased BCoV recovery by 19%, which seems to compensate for viral loss during centrifugation. Filtration time decreases by approximately 30% when using the Cp Select protocol, making this an optimal choice for building surveillance applications where a quick turnaround time is necessary.

scholarly journals First Data-Set on SARS-CoV-2 Detection for Istanbul Wastewaters in Turkey

2020 ◽  
Author(s):  
Bilge Alpaslan Kocamemi ◽  
Halil Kurt ◽  
Sabri Hacıoglu ◽  
Cevdet Yaralı ◽  
Ahmet Mete Saatci ◽  
...  
Keyword(s):  
Wastewater Treatment ◽  
Wastewater Treatment Plants ◽  
Treatment Plant ◽  
Warning Signs ◽  
Virus Concentration ◽  
Serum Samples ◽  
Data Set ◽  
Concentration Method ◽  
Diagnostic Panel ◽  
Wastewater Samples

AbstractSevere acute respiratory syndrome coronavirus 2 (SARS-CoV-2) started in Wuhan, China, in December 2019 and became a global pandemic [1]. By 26 April 2020, more than 2.9 million people were infected by SARS-CoV-2 and over 203 thousand people lost their life globally. By 26 April 2020, 107773 confirmed cases were reported in Turkey with 2706 deaths. Majority of the cases in Turkey has been observed in Istanbul. In the world, the duration of availability of SARS-CoV-2 was found to be significantly longer in stool samples than in respiratory and serum samples [2]. SARS-CoV-2 was detected in wastewaters in Australia [3], Netherlands [4], USA [5], France [6], Spain [7] and USA [8] by using different virus concentration techniques. In this work, Istanbul metropole with 65 % of Covid-19 cases was chosen as the pilot city. On the 21st of April 2020, 24-hr composite samples were collected from the Ambarli, Pasakoy and Kadikoy wastewater treatment plants (WWTP). On the 25th of April 2020, more wastewater samples were taken from Terkos, Buyukcekmece, Baltalimani and Tuzla WWTPs. These wastewater treatment plants were selected among 81 plants in Istanbul in order to take representative samples from 4 different districts of Istanbul according to the severity of Covid-19 cases, like very serious, serious, moderate and mild. Grab samples were also collected from Bagcilar and Kartal manholes located nearby the pandemic hospitals on April 21st, 2020. Polyethylene glycol 8000 (PEG 8000) adsorption [5] SARS-Cov-2 concentration method was used for SARS-CoV-2 concentration after optimization. Real time RT-PCR diagnostic panel validated by US was used to quantify SARS-CoV-2 RNA in raw sewage taken from the inlets of treatment plants and manholes. Five samples out of seven from wastewater and all samples from manholes were tested positive. SARS-CoV-2 in raw sewage from Ambarli, Pasakoy, Kadikoy, Terkos, Buyukcekmece, Baltalimani and Tuzla WWTPs were found as 8.26×103, 1.80×104, ND, ND, 3.73×103, 4.95×103, 2.89×103, respectively. The Bagcilar and Kartal manholes nearby pandemic hospitals exhibited 4.49×104 and 9.33×104, respectively. SARS-CoV-2 virus titers of manhole were higher than those of inlet of WWTPs. The observed copy numbers were presented against the number of Covid-19 cases coming to the WWTP per treatment plant capacity. Quantitative measurements of SARS-CoV-2 in wastewater can be used as a tool in wastewater-based epidemiology (WBE) and it can provide information about SARS-CoV-2 distribution in wastewater of various districts of Istanbul which exhibit different scores of Covid-19 cases. The distribution of epidemy was followed not only with blood test but with wastewater monitoring. This may allow us to identify the districts not exhibiting many Covid-19 cases, but under high risk. Continuous monitoring of wastewater for SARS-Cov-2 may provide an early warning signs before an epidemy starts in case of infection resurge.

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