Abstract

Cytomegalovirus (CMV) reactivation in the colon is common in patients with severe ulcerative colitis (UC). Ganciclovir (GCV) resistance conferring CMV UL97 gene mutations have been reported in recent years. However, the prevalence of UL97 gene mutations in GCV-naive CMV infection in the colon remains unknown. We investigated the prevalence of CMV UL97 gene mutations in patients with colonic CMV infection associated with or without UC. Twenty-two GCV-naive patients with colonic CMV infection, 15 with UC and 7 with other diseases, were enrolled. Frozen biopsy samples or formalin-fixed paraffin-embedded samples were used for nested polymerase chain reaction (PCR) amplification of the UL97 gene. Sanger DNA sequencing was performed. In comparison with AD169 reference strain, natural polymorphisms were frequently detected in codons N68D (100%), I244V (100%), and D605E (86.4%). Seven polymorphisms were detected infrequently (< 10%) outside the kinase domain. However, no known GCV resistance mutations were found. There seemed to be no difference between the ratio of polymorphisms in patients with and without UC. In conclusions, we did not detect UL97 gene mutations associated with GCV resistance in GCV-naive patients with or without UC. Consistent with previous reports, D605E polymorphism may be used as a genetic marker for CMV in East Asian countries.

Highlights

  • Cytomegalovirus (CMV) reactivation in the colon is common in patients with severe ulcerative colitis (UC)

  • Among the UC patients, steroid therapy was used for five patients (33.3%), immunosuppressive therapy was used for seven patients (46.7%), anti-TNF therapy was used for five patients (33.3%), and 5-ASA was used for 10 patients (66.7%)

  • We investigated the prevalence of CMV UL97 gene mutations and polymorphisms in Japanese patients with CMV infection in the colon for the first time

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Summary

Introduction

Cytomegalovirus (CMV) reactivation in the colon is common in patients with severe ulcerative colitis (UC). Abbreviations CMV Cytomegalovirus UC Ulcerative colitis GCV Ganciclovir HIV Human immunodeficiency virus PCR Polymerase chain reaction FFPE Formalin-fixed paraffin-embedded DNA Deoxyribonucleic acid NGS Next-generation sequencing. Well characterized GCV resistance mutations at UL97 codons 460, 520, and 590–607 impair the phosphorylation of GCV that is necessary for its antiviral activity, presumably by altering substrate r­ ecognition[14,15,16]. Despite their lack of association with GCV resistance, polymorphism likely to be related to regionality have been ­reported[17,18]

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