مطالعه مولکولی ژن‌های Tsst، Eta و Etb در باکتری Staphylococcus aureus جدا شده از بیماران مبتلا به زردزخم در میان کودکان شهر دیوانیه عراق

نوع مقاله : مقاله پژوهشی

نویسندگان

1 گروه تجزیه و تحلیل آسیب‌شناسی، دانشکده علوم، دانشگاه القادسیه، عراق

2 گروه زیست‌شناسی، دانشکده آموزش، دانشگاه القادسیه، عراق

چکیده

هدف: Staphylococcus aureus یک پاتوژن فرصت‌طلب و همه‌کاره است که مسئول طیف وسیعی از بیماری‌های انسانی، از عفونت‌های سطحی پوست گرفته تا شرایط سیستمیک تهدیدکننده حیات است. یکی از تظاهرات بالینی رایج آن زردزخم است. عفونتی مسری در پوست که عمدتاً کودکان را تحت تأثیر قرار می‌دهد و معمولاً با S. aureus به‌عنوان عامل اصلی بیماری همراه است. شیوع این باکتری در عفونت‌های پوستی و بافت نرم به یک نگرانی رو‌ به ‌رشد در حوزه سلامت عمومی، به‌ویژه در جمعیت‌های کودکان و مناطقی با دسترسی محدود به خدمات درمانی پیشرفته تبدیل شده است. مطالعه حاضر با هدف بررسی حضور مولکولی ژن‌های tsst، eta و etb در جدایه‌های S. aureus به‌دست‌آمده از کودکان مبتلا به زردزخم در شهر دیوانیه عراق انجام شد.
مواد و روش‌ها: این مطالعه بر روی جدایه‌های بالینی به‌دست‌آمده از کودکان مبتلا به زردزخم که به درمانگاه‌های پوست در شهر دیوانیه مراجعه کرده بودند انجام شد. در مجموع ۲۱ نمونه سواب از ضایعات پوستی آلوده با استفاده از سواب‌های پنبه‌ای استریل جمع‌آوری گردید. DNA ژنومی از جدایه‌های تأییدشده S. aureus استخراج شد. برای تأیید هویت جدایه‌ها، PCR هدفمند بر روی ژن 16S rRNA انجام گرفت. حضور ژن‌های tsst، eta و etb با استفاده از PCR معمولی و پرایمرهای اختصاصی بررسی شد.
نتایج: تمام جدایه‌های S. aureus باند مشخصی به اندازه ۱۵۰۰ جفت باز در PCR مربوط به ژن 1616S rRNA نشان دادند که بیانگر موفقیت‌آمیز بودن تکثیر این ژن هدف بود. این نتیجه همچنین کیفیت بالای DNA باکتریایی استخراج‌شده و اختصاصی بودن پرایمرهای مورد استفاده را نشان داد. ماهیت حفاظت‌شده ژن 16S rRNA آن را به یک نشانگر مولکولی قابل اعتماد برای شناسایی باکتری تبدیل کرده است. علاوه بر این، حضور ژن‌های tsst-1، eta و etb با استفاده از PCR بررسی شد و هر ژن الگوی تکثیر خاصی از خود نشان داد. از میان ۲۱ جدایه بالینی، ۱۱ مورد (۵۲.۴٪) برای ژن tsst-1 مثبت بودند. ژن‌های eta و etb نیز شناسایی شدند، اما با فراوانی‌های متفاوت، که نشان‌دهنده توزیع متفاوت این عوامل ویرولانس در بین جدایه‌هاست.
نتیجه‌گیری: این مطالعه حضور مولکولی ژن‌های ویرولانس کلیدی tsst-1، eta و etb را در جدایه‌های S. aureus از موارد زردزخم در کودکان برجسته می‌کند. شناسایی این ژن‌های سمی، به‌ویژه شیوع بالای tsst-1، پتانسیل بیماری‌زایی سویه‌های در گردش را برجسته کرده و اهمیت غربالگری مولکولی در تشخیص‌های بالینی و راهبردهای کنترل عفونت را تقویت می‌کند.

کلیدواژه‌ها

عنوان مقاله [English]

Molecular investigation of Tsst, Eta, and Etb Genes in Staphylococcus aureus isolated from impetigo patients among children in Al-Diwaniyah city, Iraq

نویسندگان [English]

  • Amaar Al-Jubouri 1
  • Majid Al-Shibly 2
1 Department of Pathological Analysis, College of Science, University of Al-Qadisiyah, Iraq
2 Department of Biology, College of Education, University of Al-Qadisiyah, Iraq
چکیده [English]

Objective
Staphylococcus aureus is a versatile and opportunistic pathogen responsible for a broad spectrum of human diseases ranging from superficial skin infections to life-threatening systemic situations. Among its many clinical manifestations, impetigo a contagious skin infection primarily affecting children is commonly related with S. aureus as a principal causative agent. The prevalence of S. aureus in skin and soft tissue infections has become a growing public health concern, especially in pediatric populations and in regions with restricted access to advanced healthcare resources. The current investigation aimed to investigate the molecular presence of the tsst, eta, and etb genes in S. aureus isolates achieved from children diagnosed with impetigo in Al-Diwaniyah City,. By combining 16S rRNA gene amplification with virulence gene profiling, this research contributes to a deeper understanding of the molecular epidemiology and pathogenic traits of S. aureus in a vulnerable patient population.

Materials and methods
This investigation was managed on clinical isolates achieved from children diagnosed with impetigo and attending dermatology clinics in Al-Diwaniyah City, Iraq. Totally 21 swab samples were gathered from infected skin lesions applying sterile cotton swabs. Genomic DNA was extracted from affirmed S. aureus isolates. To affirm the identity of the isolates, PCR was carried out targeting the 16S rRNA gene. The presence of tsst, eta, and etb genes was identified by conventional PCR applying gene-specific primers.
Results
All Staphylococcus aureus isolates demonstrated a distinct 1,500 bp band upon PCR amplification of the 16S rRNA gene, affirming successful amplification of the target gene. This result also indicated the high quality of the extracted bacterial DNA and the specificity of the primers applied. The conserved nature of the 16S rRNA gene reinforces its reliability as a molecular marker for bacterial identification. In addition, the presence of exfoliative toxin genes (eta, etb, and tsst-1) was evaluated applying PCR. Each gene generated a distinct amplification pattern. Among the 21 clinical isolates, 11 were positive for the tsst-1 gene. The eta and etb genes were also detected but in varying frequencies, suggesting differential distribution of these virulence factors among the isolates.

Conclusions
This investigation highlights the molecular presence of key virulence genes (tsst-1, eta, and etb) in Staphylococcus aureus isolates from impetigo cases in children. The detection of these toxin genes, particularly the high prevalence of tsst-1, underscores the pathogenic potential of circulating strains and reinforces the importance of molecular screening in clinical diagnostics and infection control strategies.

کلیدواژه‌ها [English]

  • Children
  • impetigo
  • PCR
  • Staphylococcus aureus
  • virulence genes

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مجله بیوتکنولوژی کشاورزی
دوره 17، شماره 2
اردیبهشت 1404
صفحه 325-342

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