جداسازی و بررسی خصوصیات باکتریوفاژ لیتیک علیه باکتری استاندارد کلبسیلا پنومونیه و اثر ضدباکتریایی آن روی جدایه های بالینی سودوموناس آئروژینوزا و استافیلوکوکوس اورئوس

نوع مقاله : مقاله پژوهشی

نویسندگان

دانشگاه مراغه

چکیده
مقدمه: با توجه به بروز سویه­های مقاوم به آنتی­بیوتیک باکتری‌هایی مانند کلبسیلا پنومونیه، استفاده از پتانسیل ضدباکتریایی باکتریوفاژها به­ عنوان یک رهیافت­ موثر در کنترل عفونت­ها مورد توجه قرار گرفته است.

مواد و روش­ها: پس از جداسازی باکتریوفاژ لیتیک (PKpMa1/19) علیه باکتری کلبسیلا پنومونیه PTCC 1290 از فاضلاب شهری مراغه با روش آگار دولایه، تیتراسیون، خالص سازی و غنی سازی باکتریوفاژ انجام شد. خصوصیات شکلی PKpMa1/19 با استفاده از میکروسکوپ الکترونی تعیین شد. بررسی خصوصیات رشد یک مرحله­ای، دوره نهفته، سایز انفجاری، میزان پایداری و در نهایت طیف میزبانی فاژPKpMa1/19 با استفاده از روش نقطه­ای انجام شد.

نتایج: باکتریوفاژ با خصوصیات شکلی مشابه خانواده تکتی ویریده جداسازی و خالص سازی گردید. دوره نهفته و سایز انفجاری باکتریوفاژ PKpMa1/19 به ترتیب 20 دقیقه و/cell PFU 311 تعیین شد. اثر لیتیک مناسب باکتریوفاژ بین دماهای 22- و 37 درجه سلسیوس مشخص گردید. پایداری فاژی در10-4 pH= حفظ گردید. از نظر طیف میزبانی، PKpMa1/19 فقط علیه یکی از 20 جدایه کلبسیلا پنومونیه (5 درصد جدایه های کلبسیلا) اثر ضدباکتریایی داشت ولی روی 10 جدایه سودوموناس آئروژینوزا (50 درصد جدایه های سودوموناس) و 16 جدایه استافیلوکوکوس آرئوس (80 درصد از جدایه های استافیلوکوکوس) اثر لیزکنندگی داشت.

بحث و نتیجه­ گیری: با وجود اثرات لیتیک مناسب و نیز پایداری خوب فاژ PKpMa1/19 در برابر عوامل محیطی مطالعه شده، در صورت استفاده از آن به عنوان کاندیدای فاژدرمانی، تعیین طیف میزبانی و ارزیابی اثربخشی آن علیه سویه های باکتریایی مسبب عفونت و همچنین بررسی کاملتر خصوصیات مولکولی این فاژ ضروری می باشد.

کلیدواژه‌ها

موضوعات

عنوان مقاله English

antibiotic resistance, bacteriophage, Klebsiella pneumonia, phage therapy

نویسندگان English

Raheleh Majdani
Neda Feizipour
Hamideh Lamacan
University of Maragheh
چکیده English

Introduction: Due to the emergence of antibiotic-resistant strains of bacteria such as Klebsiella pneumoniae, the use of antibacterial potential of bacteriophages has been considered as an effective approach in controlling the infections.
Material and Method: After isolating lytic bacteriophage (PKpMa1/19) against K.pneumoniae PTCC1290 from Maragheh urban sewage by agar bilayer method, purification, titration and enrichment of the bacteriophage were performed. Morphological characteristics of PKpMa1/19 were determined using electron microscopy. One step growth, latent period, burst size, stability and finally host range of the bacteriophage was evaluated using spot test.
Results: A bacteriophage with the morphological features of Tectiviridae family was isolated and purified. Its latent period and burst size were 20 min and 311 PFU respectively. Appropriate lytic effect of the bacteriophage was shown between -22 and 37 . Stability of the phage was high in pH= 4-10. The phage (PKpMa1/19), was effective against one of the 20 clinical isolates of K.pneumoniae (5% of K. pneumoniae isolates), but it showed antibacterial effect on 10 Pseudomonas aeroginosa ( 50% of P. aeruginosa isolates) and 16 Staphylococcus aureus isolates (80% of S. aureus isolates).
Discussion and Conclusion: In spite of appropriate lytic effect and also, high stability to environmental conditions, Due to its narrow host range to K. pneumoniae clinical isolates, it needs to exactly determination of its host range and efficacy of the phage against problematic strains of bacteria for using it as phage therapy candidate. Detailed survey of molecular characteristics of the bacteriophage is necessary.

کلیدواژه‌ها English

antibiotic resistance
bacteriophage
Klebsiella pneumonia
phage therapy
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  • تاریخ دریافت 18 آبان 1404
  • تاریخ پذیرش 18 اردیبهشت 1405
  • تاریخ انتشار 01 خرداد 1405