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Armoon, Mohammad, Babapour, Ebrahim, Mirnejad, Reza, Babapour, Mahshad, taati moghadam, majid. (1403). Evaluation of icaA and icaD genes involved in biofilm formation in Staphylococcus aureus isolates from clinical sources using Reverse Transcriptase PCR. سامانه مدیریت نشریات علمی, 79(6), 1329-1335. doi: 10.32592/ARI.2024.79.6.1329
Mohammad Armoon; Ebrahim Babapour; Reza Mirnejad; Mahshad Babapour; majid taati moghadam. "Evaluation of icaA and icaD genes involved in biofilm formation in Staphylococcus aureus isolates from clinical sources using Reverse Transcriptase PCR". سامانه مدیریت نشریات علمی, 79, 6, 1403, 1329-1335. doi: 10.32592/ARI.2024.79.6.1329
Armoon, Mohammad, Babapour, Ebrahim, Mirnejad, Reza, Babapour, Mahshad, taati moghadam, majid. (1403). 'Evaluation of icaA and icaD genes involved in biofilm formation in Staphylococcus aureus isolates from clinical sources using Reverse Transcriptase PCR', سامانه مدیریت نشریات علمی, 79(6), pp. 1329-1335. doi: 10.32592/ARI.2024.79.6.1329
Armoon, Mohammad, Babapour, Ebrahim, Mirnejad, Reza, Babapour, Mahshad, taati moghadam, majid. Evaluation of icaA and icaD genes involved in biofilm formation in Staphylococcus aureus isolates from clinical sources using Reverse Transcriptase PCR. سامانه مدیریت نشریات علمی, 1403; 79(6): 1329-1335. doi: 10.32592/ARI.2024.79.6.1329

Evaluation of icaA and icaD genes involved in biofilm formation in Staphylococcus aureus isolates from clinical sources using Reverse Transcriptase PCR

Archives of Razi Institute
مقاله 25، دوره 79، شماره 6، بهمن و اسفند 2024، صفحه 1329-1335    اصل مقاله (330.95 K)
نوع مقاله: Original Articles
شناسه دیجیتال (DOI): 10.32592/ARI.2024.79.6.1329
نویسندگان
Mohammad Armoon1؛ Ebrahim Babapour* 1؛ Reza Mirnejad2؛ Mahshad Babapour1؛ majid taati moghadam3
1Biotechnology Research Center, Karaj Branch, Islamic Azad University, Karaj, Iran
2Molecular Biology Research Center, Systems Biology and Poisonings Institute, Baqiyatallah University of Medical Sciences, Tehran, Iran
3Department of Microbiology, Faculty of Medicine, Guilan University of Medical Sciences, Rasht, Iran
چکیده
Staphylococcus aureus is known for its ability to create biofilm, which helps bacteria attach to various surfaces and poses a challenge for treatment. Biofilm formation relies on the icaABCD operon, with the icaA and icaD genes having the key role in this complex process. The aim of the study is to investigate the role of these genes in the biofilm formation of S. aureus isolates sourced from clinical settings. We collected 100 S. aureus isolates from clinical sources, then extracted DNA and RNA using a commercial kit from Kiagen Co. To transcribe the RNA samples into cDNA, we used a commercial kit from Kiagen Co. We measured the ability to produce phenotypic and molecular biofilm formation using the microtiter plate method and PCR, respectively. We were determining the expression of icaA and icaD genes via RT-PCR (Reverse transcription polymerase chain reaction). Ninety-five percent (95%) of the isolates were able to produce biofilm, with 16 (16%) producing weak, 64 (64%) producing medium, and 15 (15%) producing strong biofilms. Additionally, 72 (72%) of the isolates had the icaA gene, while 58 (58%) carried the icaD gene. Out of these isolates, 70 (97.2%) isolates expressed the icaA gene, and 53 (73.6%) isolates expressed the icaD gene. Four isolates (5.5%) that had the icaA gene but lacked the icaD gene did not form biofilm. One strain did not express either of the genes. The presence of either the icaA or icaD gene is crucial for the development of biofilm. However, further investigation is needed to fully understand the formation of biofilm.
کلیدواژه‌ها
Biofilm؛ Staphylococcus aureus؛ icaA gene؛ icaD gene؛ Reverse Transcriptase PCR
عنوان مقاله [English]
ارزیابی ژن‌های icaA و icaD دخیل در تشکیل بیوفیلم در جدایه‌های استافیلوکوکوس اورئوس از منابع بالینی با استفاده از PCR رونوشت معکوس
چکیده [English]
استافیلوکوکوس اورئوس به دلیل توانایی خود در ایجاد بیوفیلم معروف است که به باکتری ها کمک می کند تا به سطوح مختلف بچسبند و درمان را با چالش مواجه کند. تشکیل بیوفیلم به اپرون icaABCD متکی است و ژن‌های icaA و icaD نقش کلیدی در این فرآیند پیچیده دارند. هدف از این مطالعه بررسی نقش این ژن‌ها در تشکیل بیوفیلم جدایه‌های استافیلوکوکوس اورئوس منشأ گرفته از محیط‌های بالینی است. ما 100 ایزوله استافیلوکوکوس اورئوس را از منابع بالینی جمع آوری کردیم، سپس DNA و RNA را با استفاده از کیت تجاری از شرکت کیاژن استخراج کردیم. برای رونویسی نمونه های RNA به cDNA، از کیت تجاری شرکت کیاژن استفاده کردیم. ما توانایی تولید فنوتیپی و مولکولی را اندازه گیری کردیم. تشکیل بیوفیلم به ترتیب با استفاده از روش میکروتیتر پلیت و PCR بررسی شد و تعیین بیان ژن‌های icaA و icaD از طریق RT-PCR بودیم. نود و پنج درصد (95%) از جدایه ها قادر به تولید بیوفیلم بودند که 16 (16%) ضعیف، 64 (64%) متوسط و 15 (15%) بیوفیلم قوی تولید کردند. علاوه بر این، 72 (72٪) از جدایه ها دارای ژن icaA بودند، در حالی که 58 (58٪) حامل ژن icaD بودند. از بین این جدایه ها، 70 ایزوله (97.2%) ژن icaA و 53 ایزوله (73.6%) ژن icaD را بیان کردند. چهار جدایه (5.5%) که دارای ژن icaA بودند اما فاقد ژن icaD بودند، بیوفیلم تشکیل ندادند. یک سویه هیچ یک از ژن ها را بیان نکرد. وجود ژن icaA یا icaD برای توسعه بیوفیلم بسیار مهم است. با این حال، تحقیقات بیشتری برای درک کامل تشکیل بیوفیلم مورد نیاز است.
کلیدواژه‌ها [English]
بیوفیلم, استافیلوکوکوس اورئوس, ژن icaA, ژن icaD, PCR ترانس کریپتاز معکوس
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