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.

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.

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.

Optimization of the selection of analysis methods for the determination of naturally occurring radionuclides

Kerntechnik ◽  
2008 ◽  
Vol 73 (3) ◽  
pp. 118-121
Author(s):  
T. Heinrich ◽  
L. Funke ◽  
M. Köhler ◽  
U.-K. Schkade ◽  
F. Ullrich ◽  
...  
Keyword(s):  
Naturally Occurring ◽  
Analysis Methods ◽  
Naturally Occurring Radionuclides ◽  
Selection Of

Determination of Specific Activity of Cholera Chemical Vaccine Components using Cell Culture

2020 ◽  
Vol 36 (3) ◽  
pp. 82-89
Author(s):  
O.V. Gromova ◽  
O.S. Durakova ◽  
S.V. Generalov ◽  
L.F. Livanova ◽  
O.A. Volokh
Keyword(s):  
Cell Culture ◽  
Vibrio Cholerae ◽  
Production Control ◽  
Specific Activity ◽  
Methodological Approach ◽  
Toxin Production ◽  
Submerged Cultivation ◽  
Vaccine Production ◽  
Antigenic Activity

Том 36(2020) №3 стр. 82-89; DOI 10.21519/0234-2758-2020-36-3-82-89А.В. Гаева1*, О.В. Громова1, О.С. Дуракова1, С.В. Генералов1, Л.Ф. Ливанова1, О.А. Волох1 Определение специфической активности компонентов холерной химической вакцины с использованием культуры клеток 1ФКУЗ «Российский научно-исследовательский противочумный институт «Микроб»» Федеральной службы по надзору в сфере защиты прав потребителей и благополучия человека, Саратов 410005 *[email protected] Поступила - 2019-11-26; После доработки - 2020-03-16; Принята к публикации - 2020-05-15 Список литературы Описаны методы определения динамики продукции токсинов штаммом Vibrio cholerae 569B при глубинном культивировании в биореакторе и антигенной активности специфической фракции холерогена-анатоксина по анатоксинсвязыванию с использованием клеточных культур. Показана высокая степень соответствия результатов, полученных методами, применяемыми для контроля этапов производства холерной химической вакцины и рассмотренными в данной работе. Отмечено, что применение клеточной линии СНО-К1 наиболее перспективно для замены биомоделей на промежуточных этапах контроля активных компонентов холерной химической вакцины. Разработанный методический подход впервые предлагается использовать на этапах производства холерной бивалентной химической вакцины. культура клеток, Vibrio cholerae, холерная химическая вакцина, контроль производства, холера. Vol 36(2020) N 3 p. 82-89; DOI 10.21519/0234-2758-2020-36-3-82-89A.V. Gaeva1*, O.V. Gromova1, O.S. Durakova1, S.V. Generalov1, L.F. Livanova1, O.A. Volokh1 Determination of Specific Activity of Cholera Chemical Vaccine Components using Cell Culture 1Russian Research Anti-Plague Institute «Microbe» of the Federal Service for Surveillance on Consumer Rights Protection and Human Wellbeing, Saratov, 410005 *[email protected] Received - 26.11.2019; Accepted - 15.05.2020 References The methods has been described to determine the dynamics of toxin production by the Vibrio cholerae 569B strain during submerged cultivation in bioreactor and of the antigenic activity of specific choleragen anatoxin fraction by anatoxin binding levels using cell cultures. High degree of consistency was observed between the results obtained via the method under consideration and those obtained via control methods at different stages of cholera chemical vaccine production. It was shown that the CHO-K1 cell line is the most promising substitute for biomodels at the intermediate stages of control of active cholera chemical vaccine components. The developed methodological approach was first proposed for use at the stages of cholera chemical bivalent vaccine manufacturing. cell culture, Vibrio cholerae, cholera chemical vaccine, production control, cholera.

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