Detection and subtyping of Blastocystis sp. in human and animal stool samples using high-resolution melting analysis
Abstract Objective This study aimed to investigate the prevalence and subtype distribution of the genus Blastocystis using high-resolution melting curve analysis (HRM), a novel molecular technique. Blastocystis sp., a unicellular, anaerobic, and zoonotic parasite, is commonly found in the digestive...
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BMC
2025-02-01
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| Series: | BMC Infectious Diseases |
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| Online Access: | https://doi.org/10.1186/s12879-024-10423-y |
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| author | Ghodratollah Salehi Sangani Ahmad Hosseini-Safa Bibi Razieh Hosseini Farash Pooria Salehi Sangani Mehdi Zarean Elham Moghaddas Saeid Zoghdar Moghadam Lida Jarahi Ali Gholizadeh Fariba Berenji |
| author_facet | Ghodratollah Salehi Sangani Ahmad Hosseini-Safa Bibi Razieh Hosseini Farash Pooria Salehi Sangani Mehdi Zarean Elham Moghaddas Saeid Zoghdar Moghadam Lida Jarahi Ali Gholizadeh Fariba Berenji |
| author_sort | Ghodratollah Salehi Sangani |
| collection | DOAJ |
| description | Abstract Objective This study aimed to investigate the prevalence and subtype distribution of the genus Blastocystis using high-resolution melting curve analysis (HRM), a novel molecular technique. Blastocystis sp., a unicellular, anaerobic, and zoonotic parasite, is commonly found in the digestive tracts of humans and animals. Transmission occurs via the fecal-oral route, with prevalence rates ranging from 20% in developed countries to over 60% in developing regions. Although the pathogenicity of Blastocystis remains debated, it has been detected in both symptomatic and asymptomatic individuals. Traditionally, direct microscopy with wet mounts is used for the rapid identification of Blastocystis in stool samples. However, molecular diagnostics have become essential for the accurate detection and differentiation of Blastocystis subtypes Materials and methods We collected 730 stool samples from humans and domestic animals. Initial screening for Blastocystis sp. was performed using direct microscopy. Negative samples were cultured in a two-phase culture medium and re-examined after 2–3 days. HRM analysis was then employed to identify and differentiate Blastocystis subtypes using specific primers via real-time PCR. Results Six subtypes were identified, with ST7 (30%) being the most prevalent, followed by ST3 (28%), ST2 (16%), ST1 (14%), ST5 (6%), and ST14 (6%). In humans, ST3 was the most common subtype, also found in poultry and sheep, whereas ST7 was mainly detected in domestic animals. Notably, ST1-ST3 were identified in domesticated animals, indicating a pattern of cross-species transmission. Conclusion ST3 (28%) and ST7 (30%) were the most prevalent subtypes across all samples, with distinct distributions in human and animal hosts. The HRM technique demonstrated efficiency and cost-effectiveness, providing a rapid and accurate method for Blastocystis subtype identification in developing countries, which can expedite diagnostic responses and reduce the need for sequencing. This supports the potential for HRM to improve epidemiological surveillance and understanding of cross-species transmission. |
| format | Article |
| id | doaj-art-7ac49b62e2224bccaf19de72db550a75 |
| institution | Kabale University |
| issn | 1471-2334 |
| language | English |
| publishDate | 2025-02-01 |
| publisher | BMC |
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| series | BMC Infectious Diseases |
| spelling | doaj-art-7ac49b62e2224bccaf19de72db550a752025-02-09T12:14:24ZengBMCBMC Infectious Diseases1471-23342025-02-012511810.1186/s12879-024-10423-yDetection and subtyping of Blastocystis sp. in human and animal stool samples using high-resolution melting analysisGhodratollah Salehi Sangani0Ahmad Hosseini-Safa1Bibi Razieh Hosseini Farash2Pooria Salehi Sangani3Mehdi Zarean4Elham Moghaddas5Saeid Zoghdar Moghadam6Lida Jarahi7Ali Gholizadeh8Fariba Berenji9Department of Parasitology and Mycology, Faculty of Medicine, Mashhad University of Medical SciencesDepartment of Medical Laboratory Sciences, Faculty of Allied Medicine, Iran University of Medical SciencesDepartment of Parasitology and Mycology, Faculty of Medicine, Mashhad University of Medical SciencesDepartment of Parasitology and Mycology, Faculty of Medicine, Mashhad University of Medical SciencesDepartment of Parasitology and Mycology, Faculty of Medicine, Mashhad University of Medical SciencesDepartment of Parasitology and Mycology, Faculty of Medicine, Mashhad University of Medical SciencesDepartment of Parasitology and Mycology, Faculty of Medicine, Mashhad University of Medical SciencesDepartment of Community Medicine, Faculty of Medicine, Mashhad University of Medical SciencesDepartment of Medical Laboratory Sciences, Faculty of Allied Medicine, Gonabad University of Medical SciencesDepartment of Parasitology and Mycology, Faculty of Medicine, Mashhad University of Medical SciencesAbstract Objective This study aimed to investigate the prevalence and subtype distribution of the genus Blastocystis using high-resolution melting curve analysis (HRM), a novel molecular technique. Blastocystis sp., a unicellular, anaerobic, and zoonotic parasite, is commonly found in the digestive tracts of humans and animals. Transmission occurs via the fecal-oral route, with prevalence rates ranging from 20% in developed countries to over 60% in developing regions. Although the pathogenicity of Blastocystis remains debated, it has been detected in both symptomatic and asymptomatic individuals. Traditionally, direct microscopy with wet mounts is used for the rapid identification of Blastocystis in stool samples. However, molecular diagnostics have become essential for the accurate detection and differentiation of Blastocystis subtypes Materials and methods We collected 730 stool samples from humans and domestic animals. Initial screening for Blastocystis sp. was performed using direct microscopy. Negative samples were cultured in a two-phase culture medium and re-examined after 2–3 days. HRM analysis was then employed to identify and differentiate Blastocystis subtypes using specific primers via real-time PCR. Results Six subtypes were identified, with ST7 (30%) being the most prevalent, followed by ST3 (28%), ST2 (16%), ST1 (14%), ST5 (6%), and ST14 (6%). In humans, ST3 was the most common subtype, also found in poultry and sheep, whereas ST7 was mainly detected in domestic animals. Notably, ST1-ST3 were identified in domesticated animals, indicating a pattern of cross-species transmission. Conclusion ST3 (28%) and ST7 (30%) were the most prevalent subtypes across all samples, with distinct distributions in human and animal hosts. The HRM technique demonstrated efficiency and cost-effectiveness, providing a rapid and accurate method for Blastocystis subtype identification in developing countries, which can expedite diagnostic responses and reduce the need for sequencing. This supports the potential for HRM to improve epidemiological surveillance and understanding of cross-species transmission.https://doi.org/10.1186/s12879-024-10423-yBlastocystisZoonosisSubtypeKhorasan provinceHRM |
| spellingShingle | Ghodratollah Salehi Sangani Ahmad Hosseini-Safa Bibi Razieh Hosseini Farash Pooria Salehi Sangani Mehdi Zarean Elham Moghaddas Saeid Zoghdar Moghadam Lida Jarahi Ali Gholizadeh Fariba Berenji Detection and subtyping of Blastocystis sp. in human and animal stool samples using high-resolution melting analysis BMC Infectious Diseases Blastocystis Zoonosis Subtype Khorasan province HRM |
| title | Detection and subtyping of Blastocystis sp. in human and animal stool samples using high-resolution melting analysis |
| title_full | Detection and subtyping of Blastocystis sp. in human and animal stool samples using high-resolution melting analysis |
| title_fullStr | Detection and subtyping of Blastocystis sp. in human and animal stool samples using high-resolution melting analysis |
| title_full_unstemmed | Detection and subtyping of Blastocystis sp. in human and animal stool samples using high-resolution melting analysis |
| title_short | Detection and subtyping of Blastocystis sp. in human and animal stool samples using high-resolution melting analysis |
| title_sort | detection and subtyping of blastocystis sp in human and animal stool samples using high resolution melting analysis |
| topic | Blastocystis Zoonosis Subtype Khorasan province HRM |
| url | https://doi.org/10.1186/s12879-024-10423-y |
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