شناسایی مولکولی MRSA-MDR Staphylococcus aureus جداشده از نمونه‌های شیر خام در استان بغداد

نوع مقاله : مقاله پژوهشی

نویسنده

مرکز پژوهش‌های بازار و حمایت از مصرف‌کننده، دانشگاه بغداد، بغداد، عراق.

چکیده

هدف: این مطالعه با هدف جداسازی و شناسایی باکتری Staphylococcus aureus مقاوم به متی‌سیلین (MRSA-MDR) انجام شد. این باکتری به متی‌سیلین، که آنتی‌بیوتیکی برای درمان بسیاری از عفونت‌های ناشی از Staphylococcus aureus استفاده می‌شود مقاوم است. تصور می‌شود این مقاومت از طریق کسب ژنی حاصل شده باشد که پروتئینی را رمزگذاری می‌کند که توانایی ایجاد مقاومت را دارد. این سویه به‌عنوان Staphylococcus aureus مقاوم به متی‌سیلین (MRSA-MDR) شناخته می‌شود. حساسیت این باکتری به برخی آنتی‌بیوتیک‌ها از طریق جمع‌آوری نمونه‌های شیر خام در استان بغداد و انجام تشخیص بیوشیمیایی با استفاده از سیستم API Staph و نیز بررسی برخی از ژن‌های مسئول مقاومت به متی‌سیلین مطالعه شد.
مواد و روش‌ها: پنجاه نمونه شیر خام از بازارهای مردمی و پراکنده در سراسر بغداد (نقاط فروش غیرمجاز) که شیر خام و مشتقات آن عرضه می‌شود، در سال ۲۰۲۵ جمع‌آوری گردید. برای تأیید وابستگی نمونه‌ها به Staphylococcus aureus از آزمون تشخیصی API Staph System استفاده شد. آزمون حساسیت آنتی‌بیوتیکی برای تمامی جدایه‌های شناسایی‌شده Staphylococcus aureus با روش Kirby-Bauer Disk Diffusion انجام شد که شامل ۷ نوع آنتی‌بیوتیک سیپروفلوکساسین، اریترومایسین، آزیترومایسین، تری‌متوپریم، لووفلوکساسین، متی‌سیلین و کولیستین بود. مطالعه ژنتیکی شامل شناسایی ژن‌های تشخیصی و مقاومت آنتی‌بیوتیکی (nuc، mecA، lmrS و mepA) با استفاده از واکنش زنجیره پلیمراز (PCR) بود. برای تکثیر قطعات ژنی 276 جفت‌باز برای nuc، ‏147 جفت‌باز برای mecA، ‏718 جفت‌باز برای mepA و ‏241 جفت‌باز برای lmrSاز پرایمرهای اختصاصی استفاده شد.
نتایج: یافته‌های آزمون حساسیت آنتی‌بیوتیکی نشان داد مقاومت قابل توجهی نسبت به کولیستین (CL) با 100%، اریترومایسین (E) با 5/82% و آزیترومایسین (AZM) با 80% مقاومت وجود دارد. درصد جدایه‌های مقاوم به گروه فلوروکینولون‌ها (سیپروفلوکساسین (CIP) و لووفلوکساسین (LE)) برابر با 8/25% بود. جدایه‌های S. aureus حساسیت بالایی به تری‌متوپریم (TR) با بازدهی 3/93% نشان دادند. مطالعه ژنتیکی نشان داد 32 جدایه بر اساس مشاهده نوار 276 جفت‌بازی مربوط به ژن تشخیصی nuc پس از الکتروفورز روی ژل، Staphylococcus aureus بودند. همچنین، نتایج شناسایی مولکولی ژن mecA نشان داد که این ژن در 24 مورد (75%) از جدایه‌های Staphylococcus aureus با اندازه تکثیر 147 جفت‌باز وجود داشت.
نتیجه‌گیری: نتایج این مطالعه نشان داد که از میان 50 نمونه بررسی‌شده، 32 نمونه (64%) دارای ویژگی‌های بیوشیمیایی و مورفولوژیک اختصاصی Staphylococcus aureus بودند. مصرف گسترده آنتی‌بیوتیک‌ها منجر به بروز MRSA و دیگر باکتری‌های مقاوم چنددارویی شده است. مطالعه ژنتیکی نیز شامل شناسایی ژن‌های تشخیصی و مقاومت آنتی‌بیوتیکی (nuc، mecA، lmrS و mepA) بود.

کلیدواژه‌ها

عنوان مقاله [English]

Molecular identification of MRSA-MDR Staphylococcus aureus isolated from raw milk samples in Baghdad province

نویسنده [English]

  • Adil Turki Al-Musawi
Market Research and Consumer Protection Center, University of Baghdad, Baghdad, Iraq.
چکیده [English]

Objective
This study was conducted with the aim of isolating and identifying methicillin-resistant MRSA-MDR Staphylococcus aureus a bacterium that is known to be resistant to methicillin, an antibiotic used to treat many infections caused by Staphylococcus aureus. This strain is known as methicillin-resistant Staphylococcus aureus (MRSA-MDR). The sensitivity of this bacterium to some antibiotics was studied by collecting samples of raw milk in Baghdad Governorate after its biochemical diagnosis using the API Staph System and verifying some of the genes responsible for resistance to the methicillin.
Materials and methods
Fifty raw milk samples were sampled from random popular markets spread throughout Baghdad (unlicensed points of sale) where raw milk and its derivatives are sold for the year 2025. Diagnostic tests were conducted using the API Staph System test to confirm their affiliation to Staph.aureus specifically. The antibiotic susceptibility test was conducted to all identified Staph.aureus isolates by Kirby-Bauer Disk Diffusion technique employing 7 kinds of antibiotics Ciproflxacin, Erythromycin, Azithromycin Trimethoprim, Levofloxacin, Methicillin and Colistin. The genetic study included the detection of different identification and antibiotic-resistance genes (nuc, mecA, lmrS, and mepA) using the polymerase chain reaction (PCR) technique. For amplifying fragments of 276 bp for nuc, 147 bp for mecA, 718 bp for mepA, and 241 bp for lmrS, gene-specific primers were used.
Results
The findings of antibiotics susceptibility test showed significant degree of resistance toward Colistin (CL) 100%, Erythromycin (E) 82.50%, Azithromycin (AZM) 80%, The percentage of resistance isolates to fluoroquinolones group (Ciprofloxacin (CIP), Levofloxacin (LE)) was 25.80%. The Staph.aureus isolates recorded highly sensitivity to Trimethoprim (TR) 93.3%. The genetic study included the detection of different identification and antibiotic-resistance genes (nuc, mecA, lmrS, and mepA) using the polymerase chain reaction (PCR) technique revealed that 32 isolates were Staphylococcus aureus based on the appearance of a 276-bp band corresponding to the nuc identification gene after gel electrophoresis. Meanwhile, the results of molecular detection for mecA gene showed that it was found in 24 (75%) of Staphylococcus aureus isolates with amplicon size of 147bp.
Conclusions
The result of this study revealed that among 50 studied samples 32 (64%) isolated showed the typical biochemical and morphological characteristics that is specific to Staphylococcus aureus. The widespread of the antibiotics uses resulted in emergence of the MRSA and other multidrug resistant bacteria. The genetic study also included different identification and antibiotics resistance genes (nuc, mecA, lmrS, mepA).

کلیدواژه‌ها [English]

  • antibiotic resistance
  • API staph system
  • raw milk
  • MRSA-MDR

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مجله بیوتکنولوژی کشاورزی
دوره 18، شماره 1
فروردین 1405
صفحه 163-180

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