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

Document Type : Original Articles

Authors

1 Professor Department of Laboratory Sciences, Sirjan Faculty of Medical Sciences, Sirjan School of Medical Sciences, Sirjan, Iran

2 aInfectious Disease and Tropical Medicine Research Center, Research Institute of Cellular and Molecular Sciences in Infectious Diseases, Zahedan University of Medical Sciences, Zahedan, Iran

3 bDepartment of Parasitology and Mycology, Faculty of Medicine, Zahedan University of Medical Sciences, Zahedan, Iran

4 cDepartment of Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran

5 Department of Parasitology and Mycology, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran

6 dDepartment of Parasitology and Mycology, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran

7 fPhD Student of Medical Parasitology, Department of Parasitology and Mycology, School of Medicine Hamadan University of Medical Sciences, Hamadan, Iran

8 Research Center for Environmental Determinants of Health (RCEDH), Health Institute, Kermanshah University of Medical Sciences, Kermanshah, Iran.

9 Sirjan School of Medical Sciences, Sirjan, Iran

10.32592/ARI.2025.80.1.61

Abstract

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.

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Main Subjects

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Archives of Razi Institute
Volume 80, Issue 1
January and February 2025
Pages 61-67

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