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etemadi, soudabeh, mirahmadi, hadi, Rahmati-Balaghaleha, Mansour, darabi, enayat, zarean, mehdi, Sharifi, Yousef, Yousefnia, Hossain, parandin, fatemaeh, askari, zahra. (1403). Molecular identification and genotyping of Blastocystis spp. in children with clinical symptoms in southeast Iran using PCR-sequencing method. سامانه مدیریت نشریات علمی, 80(1), 61-67. doi: 10.32592/ARI.2025.80.1.61
soudabeh etemadi; hadi mirahmadi; Mansour Rahmati-Balaghaleha; enayat darabi; mehdi zarean; Yousef Sharifi; Hossain Yousefnia; fatemaeh parandin; zahra askari. "Molecular identification and genotyping of Blastocystis spp. in children with clinical symptoms in southeast Iran using PCR-sequencing method". سامانه مدیریت نشریات علمی, 80, 1, 1403, 61-67. doi: 10.32592/ARI.2025.80.1.61
etemadi, soudabeh, mirahmadi, hadi, Rahmati-Balaghaleha, Mansour, darabi, enayat, zarean, mehdi, Sharifi, Yousef, Yousefnia, Hossain, parandin, fatemaeh, askari, zahra. (1403). 'Molecular identification and genotyping of Blastocystis spp. in children with clinical symptoms in southeast Iran using PCR-sequencing method', سامانه مدیریت نشریات علمی, 80(1), pp. 61-67. doi: 10.32592/ARI.2025.80.1.61
etemadi, soudabeh, mirahmadi, hadi, Rahmati-Balaghaleha, Mansour, darabi, enayat, zarean, mehdi, Sharifi, Yousef, Yousefnia, Hossain, parandin, fatemaeh, askari, zahra. Molecular identification and genotyping of Blastocystis spp. in children with clinical symptoms in southeast Iran using PCR-sequencing method. سامانه مدیریت نشریات علمی, 1403; 80(1): 61-67. doi: 10.32592/ARI.2025.80.1.61

Molecular identification and genotyping of Blastocystis spp. in children with clinical symptoms in southeast Iran using PCR-sequencing method

Archives of Razi Institute
مقاله 7، دوره 80، شماره 1، فروردین و اردیبهشت 2025، صفحه 61-67    اصل مقاله (573.89 K)
نوع مقاله: Original Articles
شناسه دیجیتال (DOI): 10.32592/ARI.2025.80.1.61
نویسندگان
soudabeh etemadi* 1؛ hadi mirahmadi2؛ Mansour Rahmati-Balaghaleha3؛ enayat darabi4؛ mehdi zarean5؛ Yousef Sharifi6؛ Hossain Yousefnia7؛ fatemaeh parandin8؛ zahra askari9
1Professor Department of Laboratory Sciences, Sirjan Faculty of Medical Sciences, Sirjan School of Medical Sciences, Sirjan, Iran
2aInfectious Disease and Tropical Medicine Research Center, Research Institute of Cellular and Molecular Sciences in Infectious Diseases, Zahedan University of Medical Sciences, Zahedan, Iran
3bDepartment of Parasitology and Mycology, Faculty of Medicine, Zahedan University of Medical Sciences, Zahedan, Iran
4cDepartment of Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran
5Department of Parasitology and Mycology, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran
6dDepartment of Parasitology and Mycology, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran
7fPhD Student of Medical Parasitology, Department of Parasitology and Mycology, School of Medicine Hamadan University of Medical Sciences, Hamadan, Iran
8Research Center for Environmental Determinants of Health (RCEDH), Health Institute, Kermanshah University of Medical Sciences, Kermanshah, Iran.
9Sirjan School of Medical Sciences, Sirjan, Iran
چکیده
Blastocystis spp. is a zoonotic anaerobic parasite in the large intestine of humans and many vertebrates. It is mostly found in people who are in contact with animals. Current study aimed at identifying the prevalence of Blastocystis spp. and its common genotypes in children with clinical symptoms of diarrhea in the city of Zahedan, Southeast of Iran. A cross-sectional descriptive study was conducted on 60 children under ten years of age with gastrointestinal symptoms, especially diarrhea. After sample collection, stool samples were subjected to direct stool testing for initial diagnosis. After microscopic diagnosis, DNA extraction and Polymerase chain reaction (PCR) test with small subunit ribosomal RNA (SSU rRNA) gene target were performed, and PCR products were purified and sequenced. Nucleotide sequences were revised using Chromas biotechnology software version 2.4 and CLC genomic work bench software 11. Nucleotide sequences were aligned using BLAST database and compared with reference genotypes of Blastocystis spp. in gene bank. Genotyping of sequences was done using CLC genomic work bench software 11 and phylogenetic tree drawing was done using MEGA7 software with Neighbor-Joining statistical method applying Kimura 2-parameter method. Out of 60 examined cases, 5 children (8.33%) were positive by direct microscopic and PCR tests, where a 500 (479) bp fragment in the SSU-rRNA target was detected. Genetic analysis identified 4 subtypes, including subtypes 1, 2, 3, and 5. The percentage of nucleotide identity with the sequences in the gene bank was found to be between 93 and 100%. Considering the presence of subtypes 3 and 5 in the study and the evidence of their zoonotic, examining the parasite dynamics and epidemiological principles can be effective in the control strategy.
کلیدواژه‌ها
Blastocystis spp؛ children؛ PCR-sequencing؛ southeast Iran
عنوان مقاله [English]
شناسایی مولکولی و تعیین ژنوتیپ Blastocystis sp. در کودکان با علائم بالینی در جنوب شرق ایران با استفاده از روش PCR-sequencing
چکیده [English]
هدف: Blastocystis sp یک انگل بی هوازی مشترک بین انسان و دام در روده بزرگ انسان و بسیاری از مهره داران است. بیشتر در افرادی که با حیوانات در تماس هستند دیده می شود. مطالعه حاضر با هدف شناسایی شیوع Blastocystis sp. و ژنوتیپ های رایج آن در کودکان با علائم بالینی اسهال در شهر زاهدان، جنوب شرق ایران. انجام شد
مواد و روش ها: یک مطالعه توصیفی مقطعی بر روی 60 کودک زیر ده سال با علائم گوارشی به ویژه اسهال انجام شد. پس از جمع آوری نمونه، نمونه های مدفوع برای تشخیص اولیه تحت آزمایش مستقیم مدفوع قرار گرفتند. پس از تشخیص میکروسکوپی، استخراج DNA و آزمایش PCR با هدف ژن RNA زیر واحد کوچک ریبوزومی [SSU rRNA] انجام شد و محصولات PCR خالص و توالی یابی شدند. توالی‌های نوکلئوتیدی با استفاده از نرم‌افزار بیوتکنولوژی کروماس نسخه 2.4 و نرم‌افزار CLC Genomic Work bench 11 مورد بازبینی قرار گرفتند. ژنوتیپ توالی ها با استفاده از نرم افزار CLC Genomic Work bench 11 و ترسیم درخت فیلوژنتیک با استفاده از نرم افزار MEGA7 با روش آماری Neighbor-Joining با استفاده از روش 2 پارامتری کیمورا انجام شد.
یافته‌ها: از 60 مورد بررسی شده، 5 کودک [8.33 درصد] با آزمایش‌های میکروسکوپی مستقیم و PCR مثبت بودند که در آن قطعه 500 [479] جفت باز در هدف SSU-rRNA تشخیص داده شد. تجزیه و تحلیل ژنتیکی 4 زیرگروه شامل زیرگروه های 1، 2، 3 و 5 را شناسایی کرد. درصد همسانی نوکلئوتیدی با توالی های موجود در بانک ژن بین 93 تا 100 درصد بود.
نتیجه‌گیری: با توجه به وجود زیرگروه‌های 3 و 5 در مطالعه و شواهد زئونوز آنها، بررسی دینامیک انگل و اصول اپیدمیولوژیک می‌تواند در راهبرد کنترل مؤثر باشد.
کلیدواژه‌ها [English]
Blastocystis spp, کودکان, توالی یابی PCR, جنوب شرق ایران
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