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Cytomegalovirus Detection by PCR on Fresh and Formalin-Fixed Colon Tissue Biopsies of Children with Colitis: A Prospective Cohort Study

Articles InPress

Document Type : Original Article(s)

Authors

1 Department of Molecular Pathology and Cytogenetics, Children Medical Center Hospital, Tehran University of Medical Sciences, Tehran, Iran

2 Department of Pathology, School of Medicine, Urmia University of Medical Sciences, West Azarbayejan, Iran

3 Department of Pathology, Farabi Hospital, Tehran University of Medical Sciences, Tehran, Iran

4 Department of Pediatric Gastroenterology, Children Medical Center Hospital, Tehran University of Medical Sciences, Tehran, Iran

5 Department of Pediatric Gastroenterology, Guilan University of Medical Sciences, Rasht, Iran

6 Gene Therapy Research Center, Digestive Disease Research Institute, Shariati Hospital, Tehran University of Medical Sciences, Tehran, Iran

10.30476/ijms.2025.103842.3741

Abstract

Background: Diagnosis of cytomegalovirus (CMV) in biopsies relies on detecting classic viral cytopathic effects (CPE) in tissue. These effects are not always apparent, and confirmatory tests are necessary. This study aimed to compare the results of the different diagnostic tests for CMV detection in colitis, including PCR on fresh and formalin-fixed paraffin-embedded (FFPE) tissue, immunohistochemistry (IHC), histology, and plasma PCR, in association with the clinical course. 
Methods: In this prospective study, CMV-PCR was performed on fresh tissue (FT) and FFPE tissue, and IHC was conducted on colon biopsies from 153 children with colitis referred to Children Medical Center Hospital (Tehran, Iran) from 2015 to 2019. The results of different diagnostic methods were evaluated in association with the clinical and histopathological findings. 
Results: Fifty out of 153 (32%) cases had positive FT-CMV PCR. Forty of these fifty positive samples and 21 of 103 negative ones had concomitant FFPE biopsy tissue. FFPE-PCR and IHC were positive in 17 (42.5 %) and 2 (5%) out of 40 FT-PCR-positive cases, respectively. The two IHC-positive cases had positive FFPE-PCR and high CMV-DNA plasma levels and showed histologically active colitis and CPE. Remarkably, 14 (35%) cases were identified with positive FT-PCR without any evidence of colitis in histopathology. During follow-up, FT-PCR-positive inflammatory bowel disease (IBD) cases treated with antiviral drugs showed a poorer outcome than the non-treated cases (P=0.03).
Conclusion: A high positive rate was observed for both FT- and FFPE-CMV PCR, with a poor association with histology. IHC positivity was associated with high plasma CMV DNA levels and the presence of CPE. The efficacy of anti-CMV treatment in colitis cases should be verified through randomized controlled clinical trials. 

Highlights

Sajjadeh Movahedinia (Google Scholar)

Mohammad Vasei (Google Scholar)

Keywords

References
  1. Crough T, Khanna R. Immunobiology of human cytomegalovirus: from bench to bedside. Clin Microbiol Rev. 2009;22:76-98. doi: 10.1128/CMR.00034-08. PubMed PMID: 19136435; PubMed Central PMCID: PMCPMC2620639.
  2. Geris JM, Spector LG, Roesler M, Hernandez-Alvarado N, Blackstad M, Nelson HH, et al. High prevalence of asymptomatic CMV shedding in healthy children attending the minnesota state fair. J Clin Virol. 2022;148:105102. doi: 10.1016/j.jcv.2022.105102. PubMed PMID: 35158280; PubMed Central PMCID: PMCPMC8918022.
  3. Takahashi Y, Tange T. Prevalence of cytomegalovirus infection in inflammatory bowel disease patients. Dis Colon Rectum. 2004;47:722-6. doi: 10.1007/s10350-003-0117-3. PubMed PMID: 15037934.
  4. Al-Zafiri R, Gologan A, Galiatsatos P, Szilagyi A. Cytomegalovirus complicating inflammatory bowel disease: a 10-year experience in a community-based, university-affiliated hospital. Gastroenterol Hepatol (N Y). 2012;8:230-9. PubMed PMID: 22723754; PubMed Central PMCID: PMCPMC3380257.
  5. Lawlor G, Moss AC. Cytomegalovirus in inflammatory bowel disease: pathogen or innocent bystander? Inflamm Bowel Dis. 2010;16:1620-7. doi: 10.1002/ibd.21275. PubMed PMID: 20232408.
  6. Criscuoli V, Rizzuto MR, Cottone M. Cytomegalovirus and inflammatory bowel disease: is there a link? World J Gastroenterol. 2006;12:4813-8. doi: 10.3748/wjg.v12.i30.4813. PubMed PMID: 16937462; PubMed Central PMCID: PMCPMC4087614.
  7. Baniak N, Kanthan R. Cytomegalovirus Colitis: An Uncommon Mimicker of Common Colitides. Arch Pathol Lab Med. 2016;140:854-8. doi: 10.5858/arpa.2015-0176-RS. PubMed PMID: 27472242.
  8. Aomatsu T, Yoden A, Matsumoto K, Kimura E, Inoue K, Andoh A, et al. Fecal calprotectin is a useful marker for disease activity in pediatric patients with inflammatory bowel disease. Dig Dis Sci. 2011;56:2372-7. doi: 10.1007/s10620-011-1633-y. PubMed PMID: 21394462.
  9. Holtman GA, Lisman-van Leeuwen Y, Kollen BJ, Norbruis OF, Escher JC, Kindermann A, et al. Diagnostic Accuracy of Fecal Calprotectin for Pediatric Inflammatory Bowel Disease in Primary Care: A Prospective Cohort Study. Ann Fam Med. 2016;14:437-45. doi: 10.1370/afm.1949. PubMed PMID: 27621160; PubMed Central PMCID: PMCPMC5394359.
  10. McCoy MH, Post K, Sen JD, Chang HY, Zhao Z, Fan R, et al. qPCR increases sensitivity to detect cytomegalovirus in formalin-fixed, paraffin-embedded tissue of gastrointestinal biopsies. Hum Pathol. 2014;45:48-53. doi: 10.1016/j.humpath.2013.07.040. PubMed PMID: 24139212.
  11. Kou T, Nakase H, Tamaki H, Kudo T, Nishio A, Chiba T. Cytomegalovirus infection in patients with ulcerative colitis diagnosed by quantitative real-time PCR analysis. Dig Dis Sci. 2006;51:1052-5. doi: 10.1007/s10620-006-8006-y. PubMed PMID: 16865568.
  12. Okahara K, Nagata N, Shimada T, Joya A, Hayashida T, Gatanaga H, et al. Colonic cytomegalovirus detection by mucosal PCR and antiviral therapy in ulcerative colitis. PLoS One. 2017;12:e0183951. doi: 10.1371/journal.pone.0183951. PubMed PMID: 28886066; PubMed Central PMCID: PMCPMC5590814.
  13. Yoshida M, Kutsumi H, Kinoshita Y, Fujita T, Soga T, Nishimura K, et al. Cytomegalovirus enteritis in a nonimmunocompromised host: usefulness of polymerase chain reaction by using paraffin-embedded biopsy specimen for the diagnosis. Gastrointest Endosc. 1996;44:482-5. doi: 10.1016/s0016-5107(96)70107-4. PubMed PMID: 8905376.
  14. Temtem T, Whitworth J, Zhang J, Bagga B. Cytomegalovirus in pediatric inflammatory bowel disease patients with acute severe colitis. Clin Res Hepatol Gastroenterol. 2021;45:101625. doi: 10.1016/j.clinre.2021.101625. PubMed PMID: 33662784.
  15. Gupta RB, Harpaz N, Itzkowitz S, Hossain S, Matula S, Kornbluth A, et al. Histologic inflammation is a risk factor for progression to colorectal neoplasia in ulcerative colitis: a cohort study. Gastroenterology. 2007;133:1099-105. doi: 10.1053/j.gastro.2007.08.001. PubMed PMID: 17919486; PubMed Central PMCID: PMCPMC2175077.
  16. Geboes K, Riddell R, Ost A, Jensfelt B, Persson T, Lofberg R. A reproducible grading scale for histological assessment of inflammation in ulcerative colitis. Gut. 2000;47:404-9. doi: 10.1136/gut.47.3.404. PubMed PMID: 10940279; PubMed Central PMCID: PMCPMC1728046.
  17. El-Matary W, Stefanovici C, Van Caeseele P, Deora V, McCurdy J. Detection of Cytomegalovirus in Colonic Mucosa of Children With Inflammatory Bowel Disease: Inflammatory Bowel Disease. J Pediatr Gastroenterol Nutr. 2018;67:221-4. doi: 10.1097/MPG.0000000000001976. PubMed PMID: 29601437.
  18. Johnson J, Affolter K, Boynton K, Chen X, Valentine J, Peterson K. CMV Disease in IBD: Comparison of Diagnostic Tests and Correlation with Disease Outcome. Inflamm Bowel Dis. 2018;24:1539-46. doi: 10.1093/ibd/izy045. PubMed PMID: 29718356.
  19. Yadegarynia D, Tehrani S, Roohi M, Gachkar L, Nadji SA, Hashemi M, et al. Prevalence of cytomegalovirus infection in patients with ulcerative colitis: a prospective cross-sectional study in Tehran, Iran. Iran J Microbiol. 2018;10:342-7. PubMed PMID: 30675331; PubMed Central PMCID: PMCPMC6339997.
  20. Rashidi A, Vij KR, Buller RS, Wylie KM, Storch GA, DiPersio JF. Tissue polymerase chain reaction for the diagnosis of cytomegalovirus disease after allogeneic hematopoietic cell transplantation. Am J Hematol. 2017;92:E19-E20. doi: 10.1002/ajh.24609. PubMed PMID: 27859620; PubMed Central PMCID: PMCPMC8211556.
  21. Mills AM, Guo FP, Copland AP, Pai RK, Pinsky BA. A comparison of CMV detection in gastrointestinal mucosal biopsies using immunohistochemistry and PCR performed on formalin-fixed, paraffin-embedded tissue. Am J Surg Pathol. 2013;37:995-1000. doi: 10.1097/PAS.0b013e31827fcc33. PubMed PMID: 23648457.
  22. Talaulikar D, Gray JX, Shadbolt B, McNiven M, Dahlstrom JE. A comparative study of the quality of DNA obtained from fresh frozen and formalin-fixed decalcified paraffin-embedded bone marrow trephine biopsy specimens using two different methods. J Clin Pathol. 2008;61:119-23. doi: 10.1136/jcp.2006.045294. PubMed PMID: 17545562.
  23. Suarez-Lledo M, Marcos MA, Cuatrecasas M, Bombi JA, Fernandez-Aviles F, Magnano L, et al. Quantitative PCR Is Faster, More Objective, and More Reliable Than Immunohistochemistry for the Diagnosis of Cytomegalovirus Gastrointestinal Disease in Allogeneic Stem Cell Transplantation. Biol Blood Marrow Transplant. 2019;25:2281-6. doi: 10.1016/j.bbmt.2019.07.016. PubMed PMID: 31325586.
  24. Yoshino T, Nakase H, Ueno S, Uza N, Inoue S, Mikami S, et al. Usefulness of quantitative real-time PCR assay for early detection of cytomegalovirus infection in patients with ulcerative colitis refractory to immunosuppressive therapies. Inflamm Bowel Dis. 2007;13:1516-21. doi: 10.1002/ibd.20253. PubMed PMID: 17828781.
  25. Kim JW, Boo SJ, Ye BD, Kim CL, Yang SK, Kim J, et al. Clinical utility of cytomegalovirus antigenemia assay and blood cytomegalovirus DNA PCR for cytomegaloviral colitis patients with moderate to severe ulcerative colitis. J Crohns Colitis. 2014;8:693-701. doi: 10.1016/j.crohns.2013.12.014. PubMed PMID: 24405983.
  26. Tandon P, James P, Cordeiro E, Mallick R, Shukla T, McCurdy JD. Diagnostic Accuracy of Blood-Based Tests and Histopathology for Cytomegalovirus Reactivation in Inflammatory Bowel Disease: A Systematic Review and Meta-Analysis. Inflamm Bowel Dis. 2017;23:551-60. doi: 10.1097/MIB.0000000000001073. PubMed PMID: 28296820.
  27. Buck Q, Cho S, Mehta Walsh S, Schady D, Kellermayer R. Routine Histology-Based Diagnosis of CMV Colitis Was Rare in Pediatric Patients. J Pediatr Gastroenterol Nutr. 2022;75:462-5. doi: 10.1097/MPG.0000000000003528. PubMed PMID: 35706089.
  28. Kostakis ID, Cholidou KG, Vaiopoulos AG, Vlachos IS, Perrea D, Vaos G. Fecal calprotectin in pediatric inflammatory bowel disease: a systematic review. Dig Dis Sci. 2013;58:309-19. doi: 10.1007/s10620-012-2347-5. PubMed PMID: 22899243.
  29. Konikoff MR, Denson LA. Role of fecal calprotectin as a biomarker of intestinal inflammation in inflammatory bowel disease. Inflamm Bowel Dis. 2006;12:524-34. doi: 10.1097/00054725-200606000-00013. PubMed PMID: 16775498.
  30. Kambham N, Vij R, Cartwright CA, Longacre T. Cytomegalovirus infection in steroid-refractory ulcerative colitis: a case-control study. Am J Surg Pathol. 2004;28:365-73. doi: 10.1097/00000478-200403000-00009. PubMed PMID: 15104299.
  31. Kojima T, Watanabe T, Hata K, Shinozaki M, Yokoyama T, Nagawa H. Cytomegalovirus infection in ulcerative colitis. Scand J Gastroenterol. 2006;41:706-11. doi: 10.1080/00365520500408584. PubMed PMID: 16716970.
  32. Delvincourt M, Lopez A, Pillet S, Bourrier A, Seksik P, Cosnes J, et al. The impact of cytomegalovirus reactivation and its treatment on the course of inflammatory bowel disease. Aliment Pharmacol Ther. 2014;39:712-20. doi: 10.1111/apt.12650. PubMed PMID: 24506221.
  33. do Carmo AM, Santos FM, Ortiz-Agostinho CL, Nishitokukado I, Frota CS, Gomes FU, et al. Cytomegalovirus infection in inflammatory bowel disease is not associated with worsening of intestinal inflammatory activity. PLoS One. 2014;9:e111574. doi: 10.1371/journal.pone.0111574. PubMed PMID: 25387236; PubMed Central PMCID: PMCPMC4227676.
  34. Park SC, Jeen YM, Jeen YT. Approach to cytomegalovirus infections in patients with ulcerative colitis. Korean J Intern Med. 2017;32:383-92. doi: 10.3904/kjim.2017.087. PubMed PMID: 28490715; PubMed Central PMCID: PMCPMC5432807.
  35. Pillet S, Pozzetto B, Roblin X. Cytomegalovirus and ulcerative colitis: Place of antiviral therapy. World J Gastroenterol. 2016;22:2030-45. doi: 10.3748/wjg.v22.i6.2030. PubMed PMID: 26877608; PubMed Central PMCID: PMCPMC4726676.
  36. Roblin X, Pillet S, Oussalah A, Berthelot P, Del Tedesco E, Phelip JM, et al. Cytomegalovirus load in inflamed intestinal tissue is predictive of resistance to immunosuppressive therapy in ulcerative colitis. Am J Gastroenterol. 2011;106:2001-8. doi: 10.1038/ajg.2011.202. PubMed PMID: 21788989.
  37. Ciccocioppo R, Racca F, Paolucci S, Campanini G, Pozzi L, Betti E, et al. Human cytomegalovirus and Epstein-Barr virus infection in inflammatory bowel disease: need for mucosal viral load measurement. World J Gastroenterol. 2015;21:1915-26. doi: 10.3748/wjg.v21.i6.1915. PubMed PMID: 25684960; PubMed Central PMCID: PMCPMC4323471.
  38. Garrido E, Carrera E, Manzano R, Lopez-Sanroman A. Clinical significance of cytomegalovirus infection in patients with inflammatory bowel disease. World J Gastroenterol. 2013;19:17-25. doi: 10.3748/wjg.v19.i1.17. PubMed PMID: 23326158; PubMed Central PMCID: PMCPMC3545225.
  39. Wethkamp N, Nordlohne EM, Meister V, Helwig U, Respondek M. Identification of clinically relevant cytomegalovirus infections in patients with inflammatory bowel disease. Mod Pathol. 2018;31:527-38. doi: 10.1038/modpathol.2017.149. PubMed PMID: 29192648.
  40. Ozdemir B, Atay A, Kayhan MA, Ozin YO, Gokce DT, Altunsoy A, et al. Tissue quantitative RT-PCR test for diagnostic significance of cytomegalovirus infection in patients with inflammatory bowel disease and treatment response: Cytomegalovirus infection in patients with inflammatory bowel disease. Medicine (Baltimore). 2023;102:e34463. doi: 10.1097/MD.0000000000034463. PubMed PMID: 37543790; PubMed Central PMCID: PMCPMC10402982.
Iranian Journal of Medical Sciences

Articles InPress, Corrected Proof
Available Online from 17 August 2025
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  • Received: 08 September 2024
  • Revised: 18 October 2024
  • Accepted: 26 November 2024
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