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İçme-Kullanma Sularında Rotavirüs Varlığının EPA 1615 Yöntemi İle Tespitinin Uygulanabilirliği

Year 2020, Volume: 77 Issue: EK-4, 65 - 70, 01.05.2020

Abstract

Amaç: Gastroenteritler, dünya çapında özellikle beş yaş altı çocukları etkileyen ve ölümlere sebep olan önemli bir hastalık olup salgın durumunda salgın kaynağının kısa sürede ve doğru bir şekilde tespitinin yapılması halk sağlığının korunması açısından büyük önem arz etmektedir. Bu çalışmada, uluslararası standart bir yöntem olan EPA 1615 yönteminin uygulanabilirliğinin gösterilmesi amaçlanmıştır.Yöntem: Çalışmada ilk olarak, 10 L su örneği, klinik örneklerden izole edilmiş rotavirüs izolatı ile kirletilmiştir. Daha sonra kirletilmiş su örnekleri adsorbsiyon işlemine tabi tutulmuştur. Adsorbsiyon işlemi NanoCeram® Argonide, U.S.A. filtre kullanılarak gerçekleştirilmiştir. Kirletilmiş su örneği adsorbsiyon işlemi sonrası elüe edilmiştir. Elüsyon işlemi Beef Extract solüsyonu kullanılarak yapılmıştır. Elüsyon işlemi tamamlanan su örneğine organik flokülasyon işlemi uygulanmıştır. Organik flokülasyon işlemine geçmeden önce elde edilen elüatın pH değeri EPA 1615 standardında belirtilen değere ayarlanmıştır. Organik flokülasyon işlemi sonucunda elde edilen süpernatanın pH değeri tekrar ayarlanmıştır. Süpernatan, 30.000 MWCO tüpler ile santrifüj edilerek yoğunlaştırma işlemi tamamlanmıştır. Santrifügasyondan sonra elde edilen yoğunlaştırılmış su örneğine viral RNA izolasyonu uygulanmıştır. Çalışmada viral RNA izolasyonu, QIAamp Viral RNA Mini Kit Qiagen, Crawley-UK ile EPA 1615 standardının tanımladığı şekilde gerçekleştirilmiştir. İzole edilen viral RNA örneğinin amplifikasyon işlemi ise The RealStar® Rotavirus RT-PZR

References

  • Gajadhar AA, Allen JR. Factors contributing to the public health and economic importance of waterborne zoonotic parasites. Vet Parasitol, 2004; 126: 3-14.
  • Kittigul L, Khamoun P, Sujirarat D, Utrarachkij F, Chitpirom K, Chaichantanakit N, et al. An improved method for concentrating rotavirus from water samples. Memorias do Instituto Oswaldo Cruz, 2001; 96, 815–821.
  • Wang H, Liddell CA, Coates MM,Mooney MD, Levitz CE, Schumacher AE, et al. Global, regional, and national levels of neonatal, infant, and under-5 mortality during 1990-2013.
  • A systematic analysis for the Global Burden of Disease Study 2013. Lancet, 2014; 384: 957-79.
  • Liu L, Oza S, Hogan D, et al. Global, regional, and national causes of child mortality in 2000-13, with projections to inform post-2015 priorities: an updated systematic analysis. Lancet, 2015; 385: 430-440.
  • Ceyhan M, Alhan E, Salman N, et al. Multicenter prospective study on the burden of rotavirus gastroenteritis in Turkey, 2005-2006: a hospital- based study. J Infect Dis 2009; 200 Suppl 1: 234-8.
  • Ikner LA, Soto-Beltran M, Bright KR, New Method Using a Positively Charged Microporous Filter and Ultrafiltration for Concentration of Viruses from Tap Water. Applied and Environmental Microbiology, 2011; 77(10), 3500-3506.
  • Katayama H, Shimasaki A, Ohgaki S. Development of a Virus Concentration Method and Its Application to Detection of Enterovirus and Norwalk Virus from Coastal Seawater. Applied and Environmental Microbiology, 2002; 68(3), 1033–1039.
  • Lawrence JE, Steward GF. Purification of viruses by centrifugation. Manual of Aquatic Viral Ecology, 2010; 17, 166–181.
  • Wyn-Jones AP, Sellwood J. A Review: Enteric viruses in the aquatic environment. Journal of Applied Microbiology, 2001;91, 945-962.
  • EPA Method 1615: Measurement of Enterovirus and Norovirus Occurence in Water by Culture and RT- qPCR, 2012.
  • Shay Fout G, Cashdollar JL, Varughese EA, Parshionikar SU, Grimm AC. Measurement of enterovirus and norovirus occurrence in water by culture and RT-qPCR. I. collection of virus samples. J Vis Exp, 2015; (97): e52067, 1-7.
  • Kaynar P, Yılmaz M, Semizoğlu Ö, Eriş Zİ, Karadeniz Dursun F, Cesaretli Y ve ark. Su örneklerinde norovirüs varlığının cross-flow mikrofiltrasyon ve RT-PCR ile belirlenmesi. Turk Hij Den Biyol Derg, 2017; 74 (EK-1): 1-6.
  • Cashdollar JL, Brinkman NE, Griffin SM, McMinn BR, Rhodes ER, Varughese EA, et. al. Development and evaluation of EPA method 1615 for detection of enterovirus and norovirus in water. Appl Environ Microbiol, 2013; 79 (1): 215-23.
  • Bennett, H.B., O’Dell, H.D., Norton, G., et. al. Evaluation of a novel electropositive filter for the concentration of viruses from diverse water matrices. Water Sci Technol., 2010; 61, 317–22.
  • Cashdollar, J.L. and Dahling, D.R. Evaluation of a method to re-use electropositive cartridge filters for concentrating viruses from tap and river water. J Virol Methods, 2006; 132, 13–17.
  • Gibbons D, Rodriguez RA, Tallon L, Sobsey MD. Evaluation of positively charged alumina nanofibre cartridge filters for the primary concentration of noroviruses, adenoviruses and male-specific coliphages from seawater. J Appl Microbiol, 2010; 109, 635-41.
  • Karim MR, Rhodes ER, Brinkman N, Wymer L, Fout GS. New electropositive filter for concentrating enteroviruses and noroviruses from large volumes of water. Appl Environ Microbiol, 2009; 75: 2393-9.
  • Arya M, Shergill IS, Williamson M, Gommersal L, Arya N, Patel HRH. Basic principles of real-time quantitative PCR. Exprt Rev Mol Diagn, 2005; 5(2): 209-19.

Applicability of Rotavirus Existence in Drinking Water Samples with EPA 1615 Method

Year 2020, Volume: 77 Issue: EK-4, 65 - 70, 01.05.2020

Abstract

Objective: Gastroenteritis particularly affects children under the age of five worldwide. In case of an epidemic, determining the source of the epidemic in a short time and accurately is of great importance for the protection of the public health. In this study, it is aimed to demonstrate the applicabilitiy of the EPA 1615 method, which is an international standard method.Methods: In the study, firstly 10 L of water sample was contaminated with rotavirus isolate extracted from clinical samples. Afterwards, adsorption process was performed for the contaminated water sample. The adsorption process was achieved by using NanoCeram® Argonide, U.S.A. filter. Contaminated water sample was eluated after the adsorption process. The elution process was performed by using Beef Extract solution. Organic flocculation was applied to the eluted water sample. Before the organic flocculation process, the pH value of the eluate was adjusted to the defined value in the EPA 1615 standard. After the organic flocculation step, the pH value of the supernatant was adjusted again. Concentration process was completed by centrifugating supernatant in the 30.000 MWCO tubes. Then, viral

References

  • Gajadhar AA, Allen JR. Factors contributing to the public health and economic importance of waterborne zoonotic parasites. Vet Parasitol, 2004; 126: 3-14.
  • Kittigul L, Khamoun P, Sujirarat D, Utrarachkij F, Chitpirom K, Chaichantanakit N, et al. An improved method for concentrating rotavirus from water samples. Memorias do Instituto Oswaldo Cruz, 2001; 96, 815–821.
  • Wang H, Liddell CA, Coates MM,Mooney MD, Levitz CE, Schumacher AE, et al. Global, regional, and national levels of neonatal, infant, and under-5 mortality during 1990-2013.
  • A systematic analysis for the Global Burden of Disease Study 2013. Lancet, 2014; 384: 957-79.
  • Liu L, Oza S, Hogan D, et al. Global, regional, and national causes of child mortality in 2000-13, with projections to inform post-2015 priorities: an updated systematic analysis. Lancet, 2015; 385: 430-440.
  • Ceyhan M, Alhan E, Salman N, et al. Multicenter prospective study on the burden of rotavirus gastroenteritis in Turkey, 2005-2006: a hospital- based study. J Infect Dis 2009; 200 Suppl 1: 234-8.
  • Ikner LA, Soto-Beltran M, Bright KR, New Method Using a Positively Charged Microporous Filter and Ultrafiltration for Concentration of Viruses from Tap Water. Applied and Environmental Microbiology, 2011; 77(10), 3500-3506.
  • Katayama H, Shimasaki A, Ohgaki S. Development of a Virus Concentration Method and Its Application to Detection of Enterovirus and Norwalk Virus from Coastal Seawater. Applied and Environmental Microbiology, 2002; 68(3), 1033–1039.
  • Lawrence JE, Steward GF. Purification of viruses by centrifugation. Manual of Aquatic Viral Ecology, 2010; 17, 166–181.
  • Wyn-Jones AP, Sellwood J. A Review: Enteric viruses in the aquatic environment. Journal of Applied Microbiology, 2001;91, 945-962.
  • EPA Method 1615: Measurement of Enterovirus and Norovirus Occurence in Water by Culture and RT- qPCR, 2012.
  • Shay Fout G, Cashdollar JL, Varughese EA, Parshionikar SU, Grimm AC. Measurement of enterovirus and norovirus occurrence in water by culture and RT-qPCR. I. collection of virus samples. J Vis Exp, 2015; (97): e52067, 1-7.
  • Kaynar P, Yılmaz M, Semizoğlu Ö, Eriş Zİ, Karadeniz Dursun F, Cesaretli Y ve ark. Su örneklerinde norovirüs varlığının cross-flow mikrofiltrasyon ve RT-PCR ile belirlenmesi. Turk Hij Den Biyol Derg, 2017; 74 (EK-1): 1-6.
  • Cashdollar JL, Brinkman NE, Griffin SM, McMinn BR, Rhodes ER, Varughese EA, et. al. Development and evaluation of EPA method 1615 for detection of enterovirus and norovirus in water. Appl Environ Microbiol, 2013; 79 (1): 215-23.
  • Bennett, H.B., O’Dell, H.D., Norton, G., et. al. Evaluation of a novel electropositive filter for the concentration of viruses from diverse water matrices. Water Sci Technol., 2010; 61, 317–22.
  • Cashdollar, J.L. and Dahling, D.R. Evaluation of a method to re-use electropositive cartridge filters for concentrating viruses from tap and river water. J Virol Methods, 2006; 132, 13–17.
  • Gibbons D, Rodriguez RA, Tallon L, Sobsey MD. Evaluation of positively charged alumina nanofibre cartridge filters for the primary concentration of noroviruses, adenoviruses and male-specific coliphages from seawater. J Appl Microbiol, 2010; 109, 635-41.
  • Karim MR, Rhodes ER, Brinkman N, Wymer L, Fout GS. New electropositive filter for concentrating enteroviruses and noroviruses from large volumes of water. Appl Environ Microbiol, 2009; 75: 2393-9.
  • Arya M, Shergill IS, Williamson M, Gommersal L, Arya N, Patel HRH. Basic principles of real-time quantitative PCR. Exprt Rev Mol Diagn, 2005; 5(2): 209-19.
There are 19 citations in total.

Details

Primary Language Turkish
Journal Section Research Article
Authors

Özgül Semizoğlu This is me

Zehra İrem Eris This is me

Fatma Karadeniz Dursun This is me

Sule Senses Ergul This is me

Edibe Nurzen Namlı Bozkurt This is me

Publication Date May 1, 2020
Published in Issue Year 2020 Volume: 77 Issue: EK-4

Cite

APA Semizoğlu, Ö., Eris, Z. İ., Dursun, F. K., Ergul, S. S., et al. (2020). İçme-Kullanma Sularında Rotavirüs Varlığının EPA 1615 Yöntemi İle Tespitinin Uygulanabilirliği. Türk Hijyen Ve Deneysel Biyoloji Dergisi, 77(EK-4), 65-70.
AMA Semizoğlu Ö, Eris Zİ, Dursun FK, Ergul SS, Bozkurt ENN. İçme-Kullanma Sularında Rotavirüs Varlığının EPA 1615 Yöntemi İle Tespitinin Uygulanabilirliği. Turk Hij Den Biyol Derg. May 2020;77(EK-4):65-70.
Chicago Semizoğlu, Özgül, Zehra İrem Eris, Fatma Karadeniz Dursun, Sule Senses Ergul, and Edibe Nurzen Namlı Bozkurt. “İçme-Kullanma Sularında Rotavirüs Varlığının EPA 1615 Yöntemi İle Tespitinin Uygulanabilirliği”. Türk Hijyen Ve Deneysel Biyoloji Dergisi 77, no. EK-4 (May 2020): 65-70.
EndNote Semizoğlu Ö, Eris Zİ, Dursun FK, Ergul SS, Bozkurt ENN (May 1, 2020) İçme-Kullanma Sularında Rotavirüs Varlığının EPA 1615 Yöntemi İle Tespitinin Uygulanabilirliği. Türk Hijyen ve Deneysel Biyoloji Dergisi 77 EK-4 65–70.
IEEE Ö. Semizoğlu, Z. İ. Eris, F. K. Dursun, S. S. Ergul, and E. N. N. Bozkurt, “İçme-Kullanma Sularında Rotavirüs Varlığının EPA 1615 Yöntemi İle Tespitinin Uygulanabilirliği”, Turk Hij Den Biyol Derg, vol. 77, no. EK-4, pp. 65–70, 2020.
ISNAD Semizoğlu, Özgül et al. “İçme-Kullanma Sularında Rotavirüs Varlığının EPA 1615 Yöntemi İle Tespitinin Uygulanabilirliği”. Türk Hijyen ve Deneysel Biyoloji Dergisi 77/EK-4 (May 2020), 65-70.
JAMA Semizoğlu Ö, Eris Zİ, Dursun FK, Ergul SS, Bozkurt ENN. İçme-Kullanma Sularında Rotavirüs Varlığının EPA 1615 Yöntemi İle Tespitinin Uygulanabilirliği. Turk Hij Den Biyol Derg. 2020;77:65–70.
MLA Semizoğlu, Özgül et al. “İçme-Kullanma Sularında Rotavirüs Varlığının EPA 1615 Yöntemi İle Tespitinin Uygulanabilirliği”. Türk Hijyen Ve Deneysel Biyoloji Dergisi, vol. 77, no. EK-4, 2020, pp. 65-70.
Vancouver Semizoğlu Ö, Eris Zİ, Dursun FK, Ergul SS, Bozkurt ENN. İçme-Kullanma Sularında Rotavirüs Varlığının EPA 1615 Yöntemi İle Tespitinin Uygulanabilirliği. Turk Hij Den Biyol Derg. 2020;77(EK-4):65-70.