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Investigation of Viruses Causing Gastroenteritis by Immunochromatographic Tests and Multiplex PZR Method

Yıl 2023, , 1 - 5, 20.01.2023
https://doi.org/10.33631/sabd.1073789

Öz

Aim: Acute gastroenteritis (AGE) is a common infection all over the world, which can progress with high morbidity and mortality, especially in children, if not treated early and correctly. Rotaviruses, Adenoviruses, Noroviruses most commonly cause AGEs in childhood. While immunochromatographic (ICT) or ELISA-based methods are preferred for rapid diagnosis of viral infections, multiplex polymerase chain reaction (PCR) tests have also been used in laboratories in recent years. In this study, it is aimed to compare the results obtained by immunochromatographic and real-time PCR methods in the diagnosis of Rotavirus, Adenovirus and Norovirus.

Material and Methods: The study included 67 stool samples of a patient diagnosed with AGE who were positive for Rotavirus and/or Adenovirus by the ICT method, and 17 stool samples that were found to be negative in terms of viral, bacterial and parasitic agents by the ICT method. Rotavirus, Norovirus RNA and Adenovirus DNA were investigated in all samples using multiplex real-time PCR kit.

Results: At least one of Rota/Adeno was found to be positive in 67 of the stool samples of 84 patients included in the study. While 62 (73.8%) of the samples were Rotavirus positive with the ICT method, 49 (58.5%) were positive with the real-time PCR method. While 5 (5.9%) of the samples were found to be Adenovirus positive with the ICT method, 40 (47.6%) of them were found positive with the real-time PZR method. Norovirus was positive in 11 (13%) of the samples.


Conclusion: ICT is widely used for the diagnosis of the most common viral gastroenteritis agents Rotavirus and Adenovirus. However, as can be seen in our study; In such cases, it should not be ignored that other gastroenteritis-causing viruses, such as other Norovirus, may be responsible for the clinical picture. A significant but low level correlation was found between ICT and PCR methods in the identification of Rotavirus and Adenovirus.

Kaynakça

  • Gonzalez-Serrano L, Munoz-Algarra M, Gonzalez-Sanz R, Portero-Azorin MF, Amaro MJ. Viral gastroenteritis in hospitalized patients: Evaluation of immunochromatographic methods for rapid detection in stool samples. Journal of Clinical Virology. 2020; 128: 104420.
  • Liu L, Oza S, Hogan D, Chu Y, Perin J, Jun Zhu, et al. Global, regional, and national causes of under-5 mortality in 2000-15:an updated systematic analysis with implications for the sustainable development goals. Lancet. 2016; 388 (10063): 3027-35.
  • Fletcher M, McLaws ML, Ellis JT. Prevalence of gastrointestinal pathogens in developed and developing countries: systematic review and meta-analysis. J. Public Health Res. 2013; 2 (1): 42-53.
  • Banyai K, Estes MK, Martella V, Umesh PD. Viral gastroenteritis, Lancet. 2018; 392 175-86.
  • Sidoti F, Rittà M, Costa C, Cavallo R. Diagnosis of viral gastroenteritis: limits and potential of currently available procedures, J. Infect. Dev. 2015; 9 (6): 551-61.
  • Akhter S, Türegün B, Kıyan M, Gerçeker D, Güriz H, Şahin F. Beş yaş altı çocuklarda gastroenterite neden olan yedi farklı RNA virusunun araştırılması. Mikrobiyol Bul. 2014; 48(2): 233-41.
  • Zboromyrska Y, Vila J. Advanced PCR-based molecular diagnosis of gastrointestinal infections: challenges and opportunities. Expert review of molecular diagnostics. 2016; 16(6): 631-40.
  • Liu J, Kibiki G, Maro V, Maro A, Kumburu H, Swai N, et al. Multiplex reverse transcription PCR Luminex assay for detection and quantitation of viral agents of gastroenteritis. J Clin Virol. 2011; 50: 308-13.
  • Bennett S, Gunson RN. The development of a multiplex real-time RT-PCR for the detection of adenovirus, astrovirus, rotavirus and sapovirus from stool samples. J Virol Methods. 2016; 242: 30-4.
  • Feng W, Gu X, Sui W, Zhang M, Lu B, Wang M, et al. The application and epidemiological research of xTAG GPP multiplex PCR in the diagnosis of infectious diarrhea. Zhonghua Yi Xue Za Zhi. 2015; 95: 435-39.
  • Hyun J, Ko DH, Lee SK, KimHS, Kim JS. Evaluation of a New Multiplex Real-Time PCR assay for detecting gastroenteritis-causing viruses in stool samples. Annals of Laboratory Medicine. 2018; 38(3): 220-5.
  • Binnicker MJ. Multiplex molecular panels for diagnosis of gastrointestinal infection: performance, result interpretation, and cost-effectiveness. J Clin Microbiol. 2015; 53: 3723-8.
  • Kim J, Kim H, Kim H, Kim J, Song W, Hong Y, et al. Evaluation of an immunochromatographic assay for the rapid and simultaneous detection of rotavirus and adenovirus in stool samples. Ann Lab Med. 2014; 34: 216-22.
  • Durmaz R, Kalaycioglu AT, Acar S, Bakkaloglu Z, Karagoz A, Korukluoglu G, et al. Prevalence of rotavirus genotypes in children younger than 5 years of age before the introduction of a universal rotavirus vaccination program: report of rotavirus surveillance in Turkey. PloS one. 2014; 9(12): e113674. https://doi.org/10.1371/journal.pone.0113674
  • Altındiş M, Küçükkurt Ş, Kalaycı R, Aslan FG, Bükülmez A, Yoldaş Y. Akut gastroenteritli çocuklarda Rotavirus, enterik Adenovirus ve Norovirus sıklığı, Online Türk Sağlık Bilimleri Dergisi. 2016; 1(1): 1-12.
  • Kırdar S, Kahyaoğlu F, Yazıcı V, Aydın N. Antijen testi ile Rotavirus pozitif bulunan dışkı örneklerinde PZR ile viral gastroenterit etkenlerinin araştırılması. J Biotechnol and Strategic Health Res. 2017; 1(3): 88-93.
  • Weitzel T, Reither K, Mockenhaupt PF, Stark K, Ignatius R, Saad E, et al. Field Evaluation of a Rota- and Adenovirus ımmunochromatographic assay using stool samples from children with acute diarrhea in Ghana. Journal of Clinical Microbiology. 2007; 45(8): 2695-97.
  • https://jcm.asm.org/content/jcm/suppl/2014/12/17/JCM.02251-14. DCSupplemental/zjm999093925so1.pdf Erişim Tarihi: 08.05.2019
  • Ye S, Lambert SB, Grimwood K, Roczo-Farkas S, Nimmo GR, et al. Comparison of test specificities of commercial antigen-based assays and ın-house PZR methods for detection of Rotavirus in stool specimens. Journal of Clinical Microbiology. 2015; 53(1): 295-97.
  • Tran A, Talmud D, Lejeune B, Jovenin N, Renois F, et al. Prevalence of rotavirus, adenovirus, norovirus, and astrovirus infections and coinfections among hospitalized children in northern France. J Clin Microbiol. 2010; 48(5): 1943-6.

Gastroenterit Etkeni Virüslerin İmmünokromatografik Yöntem ve Multipleks PZR Yöntemiyle Araştırılması

Yıl 2023, , 1 - 5, 20.01.2023
https://doi.org/10.33631/sabd.1073789

Öz

Amaç: Akut gastroenterit (AGE), erken ve doğru tedavi edilmediğinde özellikle çocuklarda yüksek morbidite ve mortalite ile seyredebilen ve tüm dünyada sık görülen bir enfeksiyondur. Çocukluk döneminde en sık Rotavirüsler, Adenovirüsler, Norovirüsler AGE’lere neden olmaktadır. Viral enfeksiyonların hızlı tanısında, immünokromatografik (ICT) veya ELISA temelli yöntemler tercih edilmekte iken, son yıllarda multipleks polimeraz zincir reaksiyonu (PZR) testleri de laboratuvarlarda kullanılmaya başlanmıştır. Bu çalışmada Rotavirus, Adenovirus ve Norovirus tanısında immünokromatografik yöntem ve PZR yöntemi ile elde edilen sonuçların karşılaştırılması amaçlanmıştır.

Gereç ve Yöntemler: Çalışmaya AGE tanılı hastaların ICT yöntemi ile Rotavirus ve/veya Adenovirus pozitif olarak saptanan 67 gaita örneği ve ICT yöntemi ile viral, bakteriyel, paraziter etkenler açısından negatif bulunan 17 gaita örneği dâhil edildi. Bu örneklerde Rotavirus, Norovirus RNA’sı ile Adenovirus DNA’sı multipleks PZR kiti kullanılarak araştırıldı.


Bulgular: ICT yöntemi ile örneklerin 62’sinde (%73,8) Rotavirus pozitif iken, gerçek zamanlı PZR yöntemi ile 49’unda (%58,5) Rotavirus pozitif bulundu. ICT yöntemi ile örneklerin beşinde (%5,9) Adenovirus pozitif bulunurken, multipleks gerçek zamanlı PZR yöntemi ile 40’ında (%47,6) Adenovirus pozitif tespit edildi. Örneklerin 11’inde (%13) Norovirus pozitif saptandı.

Sonuç: En sık karşılaşılan viral gastroenterit etkenleri olan Rotavirus ve Adenovirus tanısı için çok yaygın olarak ICT kullanılmaktadır. Ancak çalışmamızda da görüldüğü üzere; bu tür olgularda Norovirüs gibi diğer gastroenterit etkeni virüslerin de klinik tablodan sorumlu olabileceği göz ardı edilmemelidir. Rotavirus ve Adenovirus tanımlamasında ICT ve PZR yöntemleri arasında anlamlı ancak düşük düzeyde korelasyon saptandı.

Kaynakça

  • Gonzalez-Serrano L, Munoz-Algarra M, Gonzalez-Sanz R, Portero-Azorin MF, Amaro MJ. Viral gastroenteritis in hospitalized patients: Evaluation of immunochromatographic methods for rapid detection in stool samples. Journal of Clinical Virology. 2020; 128: 104420.
  • Liu L, Oza S, Hogan D, Chu Y, Perin J, Jun Zhu, et al. Global, regional, and national causes of under-5 mortality in 2000-15:an updated systematic analysis with implications for the sustainable development goals. Lancet. 2016; 388 (10063): 3027-35.
  • Fletcher M, McLaws ML, Ellis JT. Prevalence of gastrointestinal pathogens in developed and developing countries: systematic review and meta-analysis. J. Public Health Res. 2013; 2 (1): 42-53.
  • Banyai K, Estes MK, Martella V, Umesh PD. Viral gastroenteritis, Lancet. 2018; 392 175-86.
  • Sidoti F, Rittà M, Costa C, Cavallo R. Diagnosis of viral gastroenteritis: limits and potential of currently available procedures, J. Infect. Dev. 2015; 9 (6): 551-61.
  • Akhter S, Türegün B, Kıyan M, Gerçeker D, Güriz H, Şahin F. Beş yaş altı çocuklarda gastroenterite neden olan yedi farklı RNA virusunun araştırılması. Mikrobiyol Bul. 2014; 48(2): 233-41.
  • Zboromyrska Y, Vila J. Advanced PCR-based molecular diagnosis of gastrointestinal infections: challenges and opportunities. Expert review of molecular diagnostics. 2016; 16(6): 631-40.
  • Liu J, Kibiki G, Maro V, Maro A, Kumburu H, Swai N, et al. Multiplex reverse transcription PCR Luminex assay for detection and quantitation of viral agents of gastroenteritis. J Clin Virol. 2011; 50: 308-13.
  • Bennett S, Gunson RN. The development of a multiplex real-time RT-PCR for the detection of adenovirus, astrovirus, rotavirus and sapovirus from stool samples. J Virol Methods. 2016; 242: 30-4.
  • Feng W, Gu X, Sui W, Zhang M, Lu B, Wang M, et al. The application and epidemiological research of xTAG GPP multiplex PCR in the diagnosis of infectious diarrhea. Zhonghua Yi Xue Za Zhi. 2015; 95: 435-39.
  • Hyun J, Ko DH, Lee SK, KimHS, Kim JS. Evaluation of a New Multiplex Real-Time PCR assay for detecting gastroenteritis-causing viruses in stool samples. Annals of Laboratory Medicine. 2018; 38(3): 220-5.
  • Binnicker MJ. Multiplex molecular panels for diagnosis of gastrointestinal infection: performance, result interpretation, and cost-effectiveness. J Clin Microbiol. 2015; 53: 3723-8.
  • Kim J, Kim H, Kim H, Kim J, Song W, Hong Y, et al. Evaluation of an immunochromatographic assay for the rapid and simultaneous detection of rotavirus and adenovirus in stool samples. Ann Lab Med. 2014; 34: 216-22.
  • Durmaz R, Kalaycioglu AT, Acar S, Bakkaloglu Z, Karagoz A, Korukluoglu G, et al. Prevalence of rotavirus genotypes in children younger than 5 years of age before the introduction of a universal rotavirus vaccination program: report of rotavirus surveillance in Turkey. PloS one. 2014; 9(12): e113674. https://doi.org/10.1371/journal.pone.0113674
  • Altındiş M, Küçükkurt Ş, Kalaycı R, Aslan FG, Bükülmez A, Yoldaş Y. Akut gastroenteritli çocuklarda Rotavirus, enterik Adenovirus ve Norovirus sıklığı, Online Türk Sağlık Bilimleri Dergisi. 2016; 1(1): 1-12.
  • Kırdar S, Kahyaoğlu F, Yazıcı V, Aydın N. Antijen testi ile Rotavirus pozitif bulunan dışkı örneklerinde PZR ile viral gastroenterit etkenlerinin araştırılması. J Biotechnol and Strategic Health Res. 2017; 1(3): 88-93.
  • Weitzel T, Reither K, Mockenhaupt PF, Stark K, Ignatius R, Saad E, et al. Field Evaluation of a Rota- and Adenovirus ımmunochromatographic assay using stool samples from children with acute diarrhea in Ghana. Journal of Clinical Microbiology. 2007; 45(8): 2695-97.
  • https://jcm.asm.org/content/jcm/suppl/2014/12/17/JCM.02251-14. DCSupplemental/zjm999093925so1.pdf Erişim Tarihi: 08.05.2019
  • Ye S, Lambert SB, Grimwood K, Roczo-Farkas S, Nimmo GR, et al. Comparison of test specificities of commercial antigen-based assays and ın-house PZR methods for detection of Rotavirus in stool specimens. Journal of Clinical Microbiology. 2015; 53(1): 295-97.
  • Tran A, Talmud D, Lejeune B, Jovenin N, Renois F, et al. Prevalence of rotavirus, adenovirus, norovirus, and astrovirus infections and coinfections among hospitalized children in northern France. J Clin Microbiol. 2010; 48(5): 1943-6.
Toplam 20 adet kaynakça vardır.

Ayrıntılar

Birincil Dil Türkçe
Konular Klinik Tıp Bilimleri
Bölüm Araştırma Makaleleri
Yazarlar

Mücahide Topçu 0000-0003-4902-4831

Mehmet Köroğlu 0000-0001-8101-1104

Özlem Aydemir 0000-0003-4533-6934

Yusuf Aydemir 0000-0003-2479-2949

Mustafa Altındiş 0000-0003-0411-9669

Bahri Elmas 0000-0001-9034-6109

Yayımlanma Tarihi 20 Ocak 2023
Gönderilme Tarihi 15 Şubat 2022
Yayımlandığı Sayı Yıl 2023

Kaynak Göster

Vancouver Topçu M, Köroğlu M, Aydemir Ö, Aydemir Y, Altındiş M, Elmas B. Gastroenterit Etkeni Virüslerin İmmünokromatografik Yöntem ve Multipleks PZR Yöntemiyle Araştırılması. SABD. 2023;13(1):1-5.