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Detection of norovirus infections in Canakkale with ELISA and RT-PCR

Year 2016, Volume: 41 Issue: 3, 533 - 538, 30.09.2016
https://doi.org/10.17826/cukmedj.237508

Abstract

Purpose: Noroviruses are important agents of gastroenteritis worldwide in every age group. There is no large countrywide dataset available for norovirus infections in Turkey. This is the first investigation of noroviruses in Çanakkale, and we aimed to compare commercial ELISA and real time RT-PCR assays in clinical use. 

Material and Methods: Fecal samples from 92 clinical gastroenteritis cases were collected and stored at -20°C. Norovirus antigen was investigated using RidaScreen ELISA and norovirus GI and GII molecular detection made with RealStar Norovirus RT-PCR kit 2.0. 

Results: We found that 16 (17.4%) samples were positive for norovirus in our study group. Using ELISA, 10 samples were positive from 92 samples. In PCR analysis, internal control amplification failed for four samples. We found 15 of 88 samples positive for real time RT-PCR analysis; of these one was GI and 14 were GII. When we accepted PCR as the reference test, the sensitivity and specificity of ELISA were calculated as 60% and 98% respectively. 

Conclusion: Single cases of norovirus infection can be seen even in rural cities like Çanakkale. There is a need to set up diagnostic capabilities for norovirus infections. ELISA has a low sensitivity, but this method is cheaper than RT-PCR and is not affected by PCR inhibitors.

References

  • Zheng DP, Ando T, Fankhauser RL, Beard RS, Glass RI, Monroe SS. Norovirus classification and proposed 2006;346:312–23. A, Virology. 2. Kirby
  • Turriza-Gómara MI. applications diagnostics :
  • interpretations. Expert Rev Anti-infective Ther.
  • ;10:423–33. and
  • Hoa Tran TN, Trainor E, Nakagomi T, Cunliffe NA, Nakagomi O. Molecular epidemiology of noroviruses associated with acute sporadic gastroenteritis in children: Global distribution of genogroups, genotypes and GII.4 variants. J Clin Virol. 2013;56:185–93.
  • Patel MM, Hall AJ, Vinjé J, Umesh DP. Noroviruses: a comprehensive review. J Clin Virol. 2009;44:1–8.
  • Uyar Y, Carhan A, Ozkaya E, Ertek M. Evaluation of laboratory diagnosis of the first norovirus outbreak in Turkey in 2008. Mikrobiyol Bul. 2008;42:607–15.
  • Albayrak N, Yagci-Caglayik D, Altaş AB, Korukluoglu G, Ertek M. Refik Saydam Hıfzıssıhha Merkezi Başkanlığı, Viroloji Referans ve Araştırma Laboratuvarı, 2009 yılı akut viral gastroenterit verilerinin değerlendirilmesi. Turk Hij ve Deney Biyol Derg. 2011;68:9–15.
  • Inan N, Kabakoğlu Ünsür E, Demirel A, Mamçu D, Sönmez E et al. Akut viral gastroenterit öntanılı vakalarda rotavirus, adenovirus ve norovirus sıklığının araştırılması. Ankem Derg. 2014;28:14–19.
  • Altindis M, Bányai K, Kalayci R, Gulamber C, Koken R, Yoldas Y et al. Frequency of norovirus in stool samples from hospitalized children due to acute gastroenteritis in Anatolia, Turkey, 2006-2007. Scand J Infect Dis. 2009;41:685–88.
  • Kapikian AZ. The discovery of the 27-nm Norwalk virus: an historic perspective. J Infect Dis. 2000;181 Suppl:S295–302.
  • Xi JN, Graham DY, Wang KN, Estes MK. Norwalk virus genome cloning and characterization. Science. 1990;250:1580–83.
  • Patel MM, Widdowson MA, Glass RI, Akazawa K, Vinje J, Parashar UD. Systematic literature review of role of noroviruses in sporadic gastroenteritis. Emer Infect Dis. 2008;14:1224–31.
  • Ozkul AA, Kocazeybek BS, Turan N, Reuter G, Bostan K, Yılmaz A et al. Frequency and phylogeny of norovirus in diarrheic children in Istanbul, Turkey J Clin Virol. 2011;51:160–64.
  • Gönen İ. Management of a large outbreak caused by norovirus and Campylobacter jejuni occurred in a rural area in Turkey. Nobel Med. 2013;9:47–51.
  • Cöl D, Biçer S, Ciler Erdağ G, Giray T, Gürol Y, Yılmaz G et al. Annual report on norovirus in children with acute gastroenteritis in 2009 and their genotypes in Turkey. Infez Med. 2013;214:261–69.
  • Mitui MT, Bozdayi G, Ahmed S, Matsumoto T, Nishizono A, Ahmed K. Detection and molecular characterization of diarrhea causing viruses in single and mixed infections in children: A comparative study between Bangladesh and Turkey. J Med Virol. 2014;86:1159–68.
  • Altay A, Bozdayı G, Meral M, Dallar Bilge Y, Dalgıç B, Özkan S et al. Investigation of norovirus infection incidence among 0-5 years old children with acute gastroenteritis admitted to two different hospitals in Ankara, Turkey. Mikrobiyol Bul. 2013;47:98–108.
  • Akhter S, Türegün B, Kiyan M, Gerçeker D, Guriz H, Şahin F. Investigation of seven different RNA viruses associated with gastroenteritis in children under five years old. Mikrobiyol Bul. 2014;48:233– 41.
  • Ahmed SF, Klena JD, Mostafa M, Dogantemur J, Middleton T, Hanson J et al. Viral gastroenteritis associated with genogroup II norovirus among U.S. military personnel in Turkey, 2009. PLoS One. 2012;7:e35791.
  • Morillo SG, Luchs A, Cilli A, Ribeiro CD, Calux SJ, Carmona Rde C et al. Norovirus 3rd Generation kit: An improvement for rapid diagnosis of sporadic gastroenteritis cases and valuable for outbreak detection. J Virol Methods. 2011;173:13–6.
  • Rovida F, Campanini G, Sarasini A, Adzasehoun KM, Piralla A, Baldanti F. Comparison of immunologic and molecular assays for the diagnosis of gastrointestinal viral infections. Diagn Microbiol Infect Dis. 2013;75:110–1.
  • Bruins MJ, Wolfhagen MJHM, Schirm J, Ruijs GJ. Evaluation of a rapid immunochromatographic test for the detection of norovirus in stool samples. Eur J Clin Microbiol Infect Dis. 2010;29:741–3.
  • Bruggink LD, Witlox KJ, Sameer R, Catton MG, Marshall JA. Evaluation of the RIDA(R)QUICK immunochromatographic norovirus detection assay using specimens from Australian gastroenteritis incidents. J Virol Methods. 2011;173:121–6.
  • Bruggink LD, Catton MG, Marshall JA. Evaluation of the Bioline Standard Diagnostics SD immunochromatographic norovirus detection kit using fecal specimens from Australian gastroenteritis incidents. Diagn Microbiol Infect Dis. 2013;76:147– 52.
  • Kele B, Lengyel G, Deak J. Comparison of an ELISA and two reverse transcription polymerase chain reaction methods for norovirus detection. Diagn Microbiol Infect Dis. 2011;70:475–8.
  • Kirby A, Gurgel RQ, Dove W, Vieria SC, Cunliffe NA, Cuevas LE. An evaluation of the RIDASCREEN and IDEIA enzyme immunoassays and the RIDAQUICK immunochromatographic test for the detection of norovirus in faecal specimens. J Clin Virol. 2010;49:254–7.
  • Mattison K, Grudeski E, Auk B, Brassard J, Charest H, Dust K et al. Analytical performance of norovirus real-time RT-PCR detection protocols in Canadian laboratories. J Clin Virol. 2011;50:109–13.
  • Duizer E, Pielaat A, Vennema H, Kroneman A, Koopmans M. Probabilities in norovirus outbreak diagnosis. J Clin Virol. 2007;40:38–42.
  • Kaplan JE, Gary GW, Baron RC, Singh N, Schonberger LB, Feldman R et al. Epidemiology of Norwalk gastroenteritis and the role of Norwalk virus in outbreaks of acute nonbacterial gastroenteritis. Ann Intern Med. 1982;96:756–61

Norovirüs enfeksiyonlarının Çanakkale'de ELİSA ve RT-PCR ile saptanması

Year 2016, Volume: 41 Issue: 3, 533 - 538, 30.09.2016
https://doi.org/10.17826/cukmedj.237508

Abstract

Amaç: Norovirüsler dünya genelinde her yaş grubunda önemli ishal etkenleridir. Türkiye'de norovirüs enfeksiyonları ile ilgili ülke genelinde geniş bir veri bulunmamaktadır. Bu çalışma Çanakkale'deki norovirüslerle ilgili ilk araştırma olup, tanıda ELISA ve real time RT-PCR testlerinin klinik kullanımlarının değerlendirilmesi amaçlanmıştır.

Gereç ve Yöntem: Klinik olarak ishali olan 92 vakadan dışkı örnekleri toplanmış olup çalışmaya kadar -20C'de saklanmıştır. Norovirüs antijeni RidaScreen ELISA kiti, GI ve GII moleküler saptaması RealStar Norovirüs RT-PCR kiti ile yapılmıştır.

Bulgular: Çalışmada 16 (%17,4) örnek norovirüs için pozitif bulunmuştur. ELISA ile 92 örnekten 10 tanesi pozitiftir. PCR çalışmasında 4 örnekte internal kontrol çalışmamış olup, 88 örnekten 15 adedinde RT-PCR'de pozitiflik saptanmıştır. Bunlardan biri GI ve ondördü GII tespit edilmiştir. PCR'ı referans test olarak aldığımızda ELISA'nın duyarlılığı %60, özgüllüğü %98 bulunmuştur.

Sonuç: Çanakkale gibi bir ilde bile norovirüs infeksiyonları birer vaka olarak bile görülebilir. Norovirüs infeksiyonlarının tanısı için altyapı oluşturulmalıdır. ELISA düşük bir duyarlılığa sahip olmakla birlikte RT-PCR'a göre ucuz olup, PCR inhibitörlerinden etkilenmemektedir.

References

  • Zheng DP, Ando T, Fankhauser RL, Beard RS, Glass RI, Monroe SS. Norovirus classification and proposed 2006;346:312–23. A, Virology. 2. Kirby
  • Turriza-Gómara MI. applications diagnostics :
  • interpretations. Expert Rev Anti-infective Ther.
  • ;10:423–33. and
  • Hoa Tran TN, Trainor E, Nakagomi T, Cunliffe NA, Nakagomi O. Molecular epidemiology of noroviruses associated with acute sporadic gastroenteritis in children: Global distribution of genogroups, genotypes and GII.4 variants. J Clin Virol. 2013;56:185–93.
  • Patel MM, Hall AJ, Vinjé J, Umesh DP. Noroviruses: a comprehensive review. J Clin Virol. 2009;44:1–8.
  • Uyar Y, Carhan A, Ozkaya E, Ertek M. Evaluation of laboratory diagnosis of the first norovirus outbreak in Turkey in 2008. Mikrobiyol Bul. 2008;42:607–15.
  • Albayrak N, Yagci-Caglayik D, Altaş AB, Korukluoglu G, Ertek M. Refik Saydam Hıfzıssıhha Merkezi Başkanlığı, Viroloji Referans ve Araştırma Laboratuvarı, 2009 yılı akut viral gastroenterit verilerinin değerlendirilmesi. Turk Hij ve Deney Biyol Derg. 2011;68:9–15.
  • Inan N, Kabakoğlu Ünsür E, Demirel A, Mamçu D, Sönmez E et al. Akut viral gastroenterit öntanılı vakalarda rotavirus, adenovirus ve norovirus sıklığının araştırılması. Ankem Derg. 2014;28:14–19.
  • Altindis M, Bányai K, Kalayci R, Gulamber C, Koken R, Yoldas Y et al. Frequency of norovirus in stool samples from hospitalized children due to acute gastroenteritis in Anatolia, Turkey, 2006-2007. Scand J Infect Dis. 2009;41:685–88.
  • Kapikian AZ. The discovery of the 27-nm Norwalk virus: an historic perspective. J Infect Dis. 2000;181 Suppl:S295–302.
  • Xi JN, Graham DY, Wang KN, Estes MK. Norwalk virus genome cloning and characterization. Science. 1990;250:1580–83.
  • Patel MM, Widdowson MA, Glass RI, Akazawa K, Vinje J, Parashar UD. Systematic literature review of role of noroviruses in sporadic gastroenteritis. Emer Infect Dis. 2008;14:1224–31.
  • Ozkul AA, Kocazeybek BS, Turan N, Reuter G, Bostan K, Yılmaz A et al. Frequency and phylogeny of norovirus in diarrheic children in Istanbul, Turkey J Clin Virol. 2011;51:160–64.
  • Gönen İ. Management of a large outbreak caused by norovirus and Campylobacter jejuni occurred in a rural area in Turkey. Nobel Med. 2013;9:47–51.
  • Cöl D, Biçer S, Ciler Erdağ G, Giray T, Gürol Y, Yılmaz G et al. Annual report on norovirus in children with acute gastroenteritis in 2009 and their genotypes in Turkey. Infez Med. 2013;214:261–69.
  • Mitui MT, Bozdayi G, Ahmed S, Matsumoto T, Nishizono A, Ahmed K. Detection and molecular characterization of diarrhea causing viruses in single and mixed infections in children: A comparative study between Bangladesh and Turkey. J Med Virol. 2014;86:1159–68.
  • Altay A, Bozdayı G, Meral M, Dallar Bilge Y, Dalgıç B, Özkan S et al. Investigation of norovirus infection incidence among 0-5 years old children with acute gastroenteritis admitted to two different hospitals in Ankara, Turkey. Mikrobiyol Bul. 2013;47:98–108.
  • Akhter S, Türegün B, Kiyan M, Gerçeker D, Guriz H, Şahin F. Investigation of seven different RNA viruses associated with gastroenteritis in children under five years old. Mikrobiyol Bul. 2014;48:233– 41.
  • Ahmed SF, Klena JD, Mostafa M, Dogantemur J, Middleton T, Hanson J et al. Viral gastroenteritis associated with genogroup II norovirus among U.S. military personnel in Turkey, 2009. PLoS One. 2012;7:e35791.
  • Morillo SG, Luchs A, Cilli A, Ribeiro CD, Calux SJ, Carmona Rde C et al. Norovirus 3rd Generation kit: An improvement for rapid diagnosis of sporadic gastroenteritis cases and valuable for outbreak detection. J Virol Methods. 2011;173:13–6.
  • Rovida F, Campanini G, Sarasini A, Adzasehoun KM, Piralla A, Baldanti F. Comparison of immunologic and molecular assays for the diagnosis of gastrointestinal viral infections. Diagn Microbiol Infect Dis. 2013;75:110–1.
  • Bruins MJ, Wolfhagen MJHM, Schirm J, Ruijs GJ. Evaluation of a rapid immunochromatographic test for the detection of norovirus in stool samples. Eur J Clin Microbiol Infect Dis. 2010;29:741–3.
  • Bruggink LD, Witlox KJ, Sameer R, Catton MG, Marshall JA. Evaluation of the RIDA(R)QUICK immunochromatographic norovirus detection assay using specimens from Australian gastroenteritis incidents. J Virol Methods. 2011;173:121–6.
  • Bruggink LD, Catton MG, Marshall JA. Evaluation of the Bioline Standard Diagnostics SD immunochromatographic norovirus detection kit using fecal specimens from Australian gastroenteritis incidents. Diagn Microbiol Infect Dis. 2013;76:147– 52.
  • Kele B, Lengyel G, Deak J. Comparison of an ELISA and two reverse transcription polymerase chain reaction methods for norovirus detection. Diagn Microbiol Infect Dis. 2011;70:475–8.
  • Kirby A, Gurgel RQ, Dove W, Vieria SC, Cunliffe NA, Cuevas LE. An evaluation of the RIDASCREEN and IDEIA enzyme immunoassays and the RIDAQUICK immunochromatographic test for the detection of norovirus in faecal specimens. J Clin Virol. 2010;49:254–7.
  • Mattison K, Grudeski E, Auk B, Brassard J, Charest H, Dust K et al. Analytical performance of norovirus real-time RT-PCR detection protocols in Canadian laboratories. J Clin Virol. 2011;50:109–13.
  • Duizer E, Pielaat A, Vennema H, Kroneman A, Koopmans M. Probabilities in norovirus outbreak diagnosis. J Clin Virol. 2007;40:38–42.
  • Kaplan JE, Gary GW, Baron RC, Singh N, Schonberger LB, Feldman R et al. Epidemiology of Norwalk gastroenteritis and the role of Norwalk virus in outbreaks of acute nonbacterial gastroenteritis. Ann Intern Med. 1982;96:756–61
There are 30 citations in total.

Details

Subjects Health Care Administration
Journal Section Research
Authors

Arif Aksu This is me

Alper Akçalı

Publication Date September 30, 2016
Acceptance Date March 27, 2016
Published in Issue Year 2016 Volume: 41 Issue: 3

Cite

MLA Aksu, Arif and Alper Akçalı. “Norovirüs enfeksiyonlarının Çanakkale’de ELİSA Ve RT-PCR Ile Saptanması”. Cukurova Medical Journal, vol. 41, no. 3, 2016, pp. 533-8, doi:10.17826/cukmedj.237508.