طراحی و توسعه‌ی Nested PCR سه‌گانه جهت تشخیص هم‌زمان Plasmodium، Babesia و Toxoplasma در فرایند انتقال خون

نوع مقاله : مقاله های پژوهشی

نویسندگان

1 دانشیار، گروه قارچ و انگل‌شناسی پزشکی، دانشکده‌ی پزشکی، دانشگاه علوم پزشکی اصفهان، اصفهان، ایران

2 دانشجوی کارشناسی ارشد، گروه قارچ و انگل‌شناسی پزشکی، دانشکده‌ی پزشکی، دانشگاه علوم پزشکی اصفهان، و سازمان انتقال خون ایران، اصفهان، ایران

چکیده

مقدمه: تشخیص و رفع عفونت‌های قابل انتقال از طریق انتقال خون از اهمیت به‌سزایی برخوردار می‌باشند. هدف از انجام این مطالعه، تشخیص Plasmodium، Toxoplasma و Babesia به طور هم‌زمان در نمونه‌های خون بود.روش‌ها: این مطالعه از نوع تجربی و توسعه‌ی روش Triplex nested-Polymerase chain reaction (Triplex nested-PCR) جهت ارتقای سلامت فرایند انتقال خون است که طی سال 1398 در اصفهان انجام گردید. پس از طراحی مطالعه و انتخاب پرایمرها، ابتدا روش مولکولی Nested PCR برای هر انگل در سطح جنس با استفاده از نمونه‌های خون با تعداد معین انگل اعمال شد. سپس، با تجمیع شرایط لازم، Nested PCR چندگانه کارسازی شد.یافته‌ها: واکنش سه‌گانه‌ی Triplex nested PCR در مقایسه با روش میکروسکوپی و Enzyme-linked immunosorbent assay (ELISA)، دارای اعتبار تشخیصی بالاتری بود؛ به گونه‌ای که مقدار حساسیت، ویژگی، ارزش اخباری مثبت و منفی برای تشخیص Plasmodium به ترتیب 100، 100، 100 و 100 درصد، برای Toxoplasma به ترتیب 100، 91، 92 و 100 درصد و برای Babesia به ترتیب 94، 100، 100 و 89 درصد ارتقا یافت. مقادیر اعتبار این آزمون برای Plasmodium، بالاترین میزان در مقایسه با دو انگل دیگر بود.نتیجه‌گیری: بر اساس نتایج این تحقیق، روش سه‌گانه‌ی (TN-PCR) Nested می‌تواند به طور هم‌زمان، یک، دو یا هر سه پاتوژن خطرناک را در نمونه‌های بیولوژیکی و همچنین، نمونه‌ی خون در فرایند انتقال خون را با دقت بالایی شناسایی کند.

کلیدواژه‌ها

عنوان مقاله [English]

A Triplex Nested Polymerase Chain Reaction for Simultaneously Diagnosis of Plasmodium, Babesia, and Toxoplasma in Blood to Promote Safety of Blood Transfusion

نویسندگان [English]

  • Abbasali Eskandarian 1
  • Alireza Najafirad 2
1 Associate Professor, Department of Parasitology and Mycology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran
2 MSc Student, Department of Parasitology and Mycology, School of Medicine, Isfahan University of Medical Sciences AND Isfahan Blood Transfusion Organization, Isfahan, Iran
چکیده [English]

Background: Diagnosis and remove of blood transmittable infections has a vital importance in blood transfusion. The aim of this study was to concurrently assess the presence of protozoan parasites plasmodium, toxoplasma, and babesia in samples of blood transfusion.Methods: In this experimental study, the development of a triplex nested-Polymerase chain reaction (PCR) for extol of blood transfusion process at 2020 in Isfahan City, Iran, was done. After selecting the genus specific primers, at the first step, nested PCRs were set up for each parasite specifically at the genus level using blood samples with a determined number of each parasite, and then multiplex nested PCR was set up.Findings: The nested PCR triple reaction (TN-PCR) has a higher diagnostic validity compared to the microscopic and enzyme-linked immunosorbent assay (ELISA) methods. Sensitivity, specificity, and positive and negative predictive values were as 100%, 100%, 100%, and 100%, respectively, for detection Plasmodium, 100%, 91%, 92%, and 100%, for Toxoplasma, and 94%, 100%, 100%, and 89% for Babesia. The validity of this test for Plasmodium was the highest compared to the other two parasites.Conclusion: Based on the results of this study, the TN-PCR method can simultaneously identify one, two or all three dangerous pathogens in biological samples as well as blood samples in the blood transfusion process.

کلیدواژه‌ها [English]

  • Plasmodium
  • Babesia
  • Toxoplasma
  • Nested polymerase chain reaction
  • Blood Transfusion

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سابقه مقاله

  • تاریخ دریافت: 18 آذر 1399
  • تاریخ بازنگری: 11 آبان 1403
  • تاریخ پذیرش: 17 فروردین 1401

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