بررسی موتاسیون های ژنUL54 مقاومت علیه گانسیکلوویر در گیرندگان پیوند کلیه و سلول های بنیادی خونساز مبتلا به عفونت سیتومگالوویروس
محورهای موضوعی : ویروس شناسیلیلا جلیل ثانی 1 , رامین یعقوبی 2 , بیتا گرامی زاده 3 , افسون افشاری 4 , محمد حسین کریمی 5
1 - دانشجوی دکترای تخصصی، گروه میکروبیولوژی، واحد شیراز، دانشگاه آزاد اسلامی، شیراز، ایران.
2 - استادیار، مرکز تحقیقات پیوند اعضا، دانشکده علوم پزشکی شیراز،شیراز، ایران
3 - استاد، مرکز تحقیقات پیوند و ترمیم اعضاء، دانشگاه علوم پزشکی شیراز، شیراز، ایران.
4 - استادیار، مرکز تحقیقات نفرو اورولوژی شیراز، دانشگاه علوم پزشکی شیراز، شیراز، ایران
5 - دانشیار، مرکز تحقیقات پیوند و ترمیم اعضاء، دانشگاه علوم پزشکی شیراز، شیراز، ایران
کلید واژه: پیوند کلیه, پیوند سلول‌های بنیادی خونساز, سیتومگالوویروس, مقاومت دارویی,
چکیده مقاله :
سابقه و هدف: مقاومت سیتومگالوویروس به گانسیکلوویر با جهش های خاص در ژنUL54 عاملی در شکست درمان و پیشرفت بیماری در گیرندگان پیوند اعضا، به ویژه پیوند کلیه و سلول های بنیادی خونساز است. این مطالعه با هدف تعیین جهش‌های منجر به مقاومت در ژن UL54 مگالوویروس انسانی انجام شد.
مواد و روش‌ها: در این مطالعه پس از غربالگری 23 گیرنده پیوند کلیه و 2 گیرنده سلول های بنیادی خونساز با تست CMV اولیه مثبت، 6 بیمار بر اساس معیارهای ورود (حداقل 2 پیگیری با نتایج Real-time PCR مثبت CMVاز پیگیری دوم به بعد) وارد مطالعه نهایی شدند. ژنUL54 با Nested-PCR تکثیر شد و محصولات PCR برای توالی یابی با استفاده از روش سنگر تعیین توالی شدند. برای تجزیه و تحلیل نتایج توالی یابی از نرم افزار فینچ (نسخه 0/4/1) استفاده شد.
یافته‌ها: پس از بررسی نتایج توالی یابی، هیچ جهش شناخته شده‌ای در 4 بیمار دریافت‌کننده پیوند کلیه مشاهده نشد. همچنین، جهش serine 882 insertion در ژن UL54 در 1 بیمار گیرنده پیوند سلول‌های بنیادی خونساز مشاهده . بررسی درخت فیلوژنی ژن UL54نشان داد که جدایه ایرانی از نظر اجدادی به 20 سویه مرجع از جمله سویه مرلین تعلق دارد.
نتیجه گیری: با توجه به اینکه ظهور جهش serine 882 insertion می تواند پتانسیل درمان را ضعیف کند و پاسخ به گانسیکلوویر را دچار مشکل کند، نظارت بر بیماران مورد نظر، تعیین بار ویروسی و ارزیابی پاسخ یا عدم پاسخ آنها به درمان بسیار مهم است.
Background & Objectives: Cytomegalovirus resistance to ganciclovir with specific mutations in the UL54 gene is a factor in treatment failure and disease progression in organ transplant recipients, particularly kidney and hematopoietic stem cell transplant recipients. This study aimed to determine the mutations leading to resistance in the UL54 gene of human cytomegalovirus.
Materials and Methods: In this study, after screening 23 kidney transplant and 2 hematopoietic stem cell transplant recipients with a positive initial CMV test, 6 patients were included in the final study based on the inclusion criteria (at least 2 available follow-ups with positive CMV Real-time PCR results). The UL54 gene was amplified using Nested-PCR, and the PCR products were then sequenced using the Sanger sequencing method. Finch software (version 1.4.0) was used to analyze the sequencing results.
Results: After reviewing the sequencing results, no known mutations were observed in 4 kidney transplant recipients. Also, a serine 882 insertion mutation in the UL54 gene was observed in 1 hematopoietic stem cell transplant recipient. Examination of the phylogenetic tree of the UL54 gene showed that the Iranian isolate ancestrally belongs to 20 reference strains, including the Merlin strain.
Conclusion: Given that the emergence of the serine 882 insertion mutation can weaken the potential for treatment and impair response to ganciclovir, it is very important to monitor the patients in question, determine the viral load, and assess their response or lack of response to treatment.
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