الكشف عن سلالات النمط الوراثي بيجينغ للمتفطرات السلية باستخدام تضخيم الدنا بالزمن الحقيقي ودرجة حرارة انفصال شريطيها
Abstract
The Beijing family of Mycobacterium tuberculosis has been reported to have exceptional capacity to spread tuberculosis and induce drug multi-resistance. A method has been developed to distinguish this family from the rest of Mycobacterium tuberculosis families through real time DNA amplification and subsequent analysis of amplicon’s melting point. Measuring the derivative of fluorescence emission with respect to temperature and plotting it against temperature unveiled the presence of a peak reflecting the amplicon's melting temperature that is diagnostic of the Beijing family (Tm: 87.23⁰C), and another one specific for the non Beijing families of strains (Tm: 82.67⁰C). This research presents an effective differentiation of bacterial strains, by applying the analysis of differential amplicon melting points.
تبين أن لعائلة سلالات البيجينغ التابعة للمتفطرات السلية قدرة مميزة على نشر مرض السل وإحداث مقاومة دوائية متعددة. تم تطوير تقنية هدفها تمييز هذه العائلة من باقي عوائل المتفطرات السلية؛ من خلال تضخيم الدنا بالزمن الحقيقي Real time PCR ودراسة درجة حرارة انصهار Melting point temperature الدنا المضخمةAmplicon . وأظهر تتبع انصهار الدنا بقياس اشتقاق الفلورة المنبعثة بالنسبة إلى متغير درجة الحرارة وجود قمة تعكس درجة حرارة انصهار الدنا المضخمة؛ التي تشخص وجود سلالة تنتمي إلى عائلة بيجينغ (درجة حرارة انصهار .2387 م)؛ وقمة أخرى خاصة بالسلالات التي لا تنتمي إلى عائلة بيجينغ (درجة حرارة انصهار .6782 م). ويمثل هذا البحث تطبيقاً فعالاً لتقنية تمييز السلالات البكتيرية الممرضة انطلاقا من تباين درجة حرارة انصهار الدنا المضخمة.
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