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Detection of Species and Molecular Typing of Leishmania in Suspected Patients by Targeting Cytochrome b Gene in Zahedan, Southeast of Iran


Affiliations
1 Infectious Diseases and Tropical Medicine Research Center, Zahedan University of Medical Sciences, Zahedan, Iran, Islamic Republic of
2 Department of Parasitology and Mycology, Zahedan University of Medical Sciences, Zahedan, Iran, Islamic Republic of
3 Department of Medical Parasitology and Mycology, School of Medicine, Qazvin University of Medical Sciences, Qazvin, Iran, Islamic Republic of
4 Department of Laboratory Sciences, Maragheh University of Medical Sciences, Maragheh, Iran, Islamic Republic of
 

Aim: Cutaneous leishmaniasis (CL) is one of the most important health problems that are capable of involving both tropical and subtropical areas, especially in Iran. This cross-sectional study aimed to differentiate the species that are able to cause CL in Zahedan city by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method.

Materials and Methods: It was conducted on 145 suspected CL patients in Zahedan city between 2014 and 2016. The smears were initially prepared, air-dried, fixed with absolute methanol, and stained with 10% Giemsa. Then, we examined the stained samples by a light microscope under 1000× magnifications. PCR assay targeted cytochrome b (cyt b) gene using LCBF1 and LCBR2 primers and the products digested by Ssp1 enzymes.

Results: From 145 suspected CL patients, 76 (52.4%) were positive in microscopic examination. In addition, we detected gene of interest (cyt b) in 98 (67.5%). The results of PCR-RFLP indicated that 53/98 (54%) cases were Leishmania major and 45/98 (46%) were Leishmania tropica, and the main species in these areas was L. major.

Conclusion: We concluded that the microscopic examination is not sensitive enough and is not able to distinguish between different Leishmania species. Instead, molecular methods like PCR-RFLP can be appropriately used with promising results.

Keywords

Cytochrome b, Leishmania major, Leishmania tropica, Polymerase Chain Reaction-Restriction Fragment Length Polymorphism.
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  • Detection of Species and Molecular Typing of Leishmania in Suspected Patients by Targeting Cytochrome b Gene in Zahedan, Southeast of Iran

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Authors

Hadi Mirahmadi
Infectious Diseases and Tropical Medicine Research Center, Zahedan University of Medical Sciences, Zahedan, Iran, Islamic Republic of
Nasrin Rezaee
Infectious Diseases and Tropical Medicine Research Center, Zahedan University of Medical Sciences, Zahedan, Iran, Islamic Republic of
Ahmad Mehravaran
Department of Parasitology and Mycology, Zahedan University of Medical Sciences, Zahedan, Iran, Islamic Republic of
Peyman Heydarian
Department of Medical Parasitology and Mycology, School of Medicine, Qazvin University of Medical Sciences, Qazvin, Iran, Islamic Republic of
Saber Raeghi
Department of Laboratory Sciences, Maragheh University of Medical Sciences, Maragheh, Iran, Islamic Republic of

Abstract


Aim: Cutaneous leishmaniasis (CL) is one of the most important health problems that are capable of involving both tropical and subtropical areas, especially in Iran. This cross-sectional study aimed to differentiate the species that are able to cause CL in Zahedan city by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method.

Materials and Methods: It was conducted on 145 suspected CL patients in Zahedan city between 2014 and 2016. The smears were initially prepared, air-dried, fixed with absolute methanol, and stained with 10% Giemsa. Then, we examined the stained samples by a light microscope under 1000× magnifications. PCR assay targeted cytochrome b (cyt b) gene using LCBF1 and LCBR2 primers and the products digested by Ssp1 enzymes.

Results: From 145 suspected CL patients, 76 (52.4%) were positive in microscopic examination. In addition, we detected gene of interest (cyt b) in 98 (67.5%). The results of PCR-RFLP indicated that 53/98 (54%) cases were Leishmania major and 45/98 (46%) were Leishmania tropica, and the main species in these areas was L. major.

Conclusion: We concluded that the microscopic examination is not sensitive enough and is not able to distinguish between different Leishmania species. Instead, molecular methods like PCR-RFLP can be appropriately used with promising results.

Keywords


Cytochrome b, Leishmania major, Leishmania tropica, Polymerase Chain Reaction-Restriction Fragment Length Polymorphism.

References