RESEARCH PAPER
Single-strand conformation polymorphism-based genetic characterization of the Cyclospora cayetanensis strains collected from different provinces in Turkey
1
Department of Parasitology, Faculty of Medicine, Kırşehir Ahi Evran University, Turkey
2
Department of Genetic, Faculty of Veterinary Medicine, Dicle University, Turkey
3
Department of Parasitology, Faculty of Medicine, Yuzuncu Yil University, Turkey
4
Department of Parasitology, Faculty Medicine, Ordu University, Turkey
Corresponding author
Muttalip Cicek
Kırşehir Ahi Evran University, Faculty of Medicine, Department of Parasitology, Kırşehir Ahi Evran Üniversitesi Tıp Fakültesi Deka, 40100, Kırşehir, Turkey
Kırşehir Ahi Evran University, Faculty of Medicine, Department of Parasitology, Kırşehir Ahi Evran Üniversitesi Tıp Fakültesi Deka, 40100, Kırşehir, Turkey
Ann Agric Environ Med. 2021;28(2):267-270
KEYWORDS
TOPICS
ABSTRACT
Introduction and objective:
Cyclospora cayetanensis, a coccidian protozoan species, has been recently found to cause diarrhea in all age groups in immunocompetent and immunocompromised individuals in most regions of the world. This study aimed to conduct the molecular detection of C. cayetanensis and to determine the genetic diversity of the 18S ribosomal RNA (rRNA) gene sequence of C. cayetanensis isolated from individuals living in different provinces in Turkey by using PCR–single-strand conformation polymorphism (SSCP).
Material and methods:
A total of 22 subjects were included in the study. Fourteen of the subjects were female and eight were male, with ages ranging between 7–65 years. Stool specimens were examined using wet mount and modified acid-fast staining methods, which revealed the presence of oocysts in the samples. The 18S rRNA ITS-1 Ccits37f-GCTTGCTATGTTTTAGCATGTGG and Ccits501r-GCACAATGAATGCACACACA gene regions were used as primers. The PCR products were analyzed by agarose gel electrophoresis and visualized on a UV transilluminator. For the SSCP, the PCR products were denatured with formamide, run for 16 h in 6% (49:1) polyacrylamide gel, and then imaged with silver staining.
Results:
SSCP assay was performed given that the DNA strands demonstrated different folds; the DNA strands contain different nucleotides based on the PCR-SSCP results for the Cyclospora strains collected in 4 provinces. Moreover, 3 different band profiles were observed in the investigated samples. A slight mutation difference was observed among the strains collected.
Conclusions:
Further comprehensive studies involving more C. cayetanensis-positive specimens and utilizing different mutation screening methods are warranted to demonstrate mutation differences in Cyclopora strains in Turkey.
Cyclospora cayetanensis, a coccidian protozoan species, has been recently found to cause diarrhea in all age groups in immunocompetent and immunocompromised individuals in most regions of the world. This study aimed to conduct the molecular detection of C. cayetanensis and to determine the genetic diversity of the 18S ribosomal RNA (rRNA) gene sequence of C. cayetanensis isolated from individuals living in different provinces in Turkey by using PCR–single-strand conformation polymorphism (SSCP).
Material and methods:
A total of 22 subjects were included in the study. Fourteen of the subjects were female and eight were male, with ages ranging between 7–65 years. Stool specimens were examined using wet mount and modified acid-fast staining methods, which revealed the presence of oocysts in the samples. The 18S rRNA ITS-1 Ccits37f-GCTTGCTATGTTTTAGCATGTGG and Ccits501r-GCACAATGAATGCACACACA gene regions were used as primers. The PCR products were analyzed by agarose gel electrophoresis and visualized on a UV transilluminator. For the SSCP, the PCR products were denatured with formamide, run for 16 h in 6% (49:1) polyacrylamide gel, and then imaged with silver staining.
Results:
SSCP assay was performed given that the DNA strands demonstrated different folds; the DNA strands contain different nucleotides based on the PCR-SSCP results for the Cyclospora strains collected in 4 provinces. Moreover, 3 different band profiles were observed in the investigated samples. A slight mutation difference was observed among the strains collected.
Conclusions:
Further comprehensive studies involving more C. cayetanensis-positive specimens and utilizing different mutation screening methods are warranted to demonstrate mutation differences in Cyclopora strains in Turkey.
ACKNOWLEDGEMENTS
The study was supported by Dicle University in Diyarbakır,
Turkey, Scientific Research Coordinatorship (DÜBAP),
Project No. 11-TF-58.
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| eISSN: | 1898-2263 |
| ISSN: | 1232-1966 |
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