Identification of Leishmania parasites in clinical samples obtained from cutaneous leishmaniasis patients using PCR-RFLP technique in endemic region, Sanliurfa province, in Turkey.
Antroponotic cutaneous leishmaniasis (ACL) is an endemic disease and one of the major health problems in Sanliurfa province located in the southeastern region of Turkey. Leishmania tropica is confirmed as the causative agent of ACL in this region. In Sanliurfa city alone, the recorded total cases of ACL were 6,817 between 2001 and 2006. We aimed to determine the effectiveness of a polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method for identification and differentiation of the Leishmania parasite in comparison to direct microscopic examination of clinical samples. The lesion exudates were collected from 51 ACL suspected patients and used for smear-slide preparations and DNA isolation. The isolated DNA was amplified by PCR, including primers selected on repetitive DNA for identification of a Leishmania subgenus, and the amplified DNA was restricted by HaeIII restriction endonuclease. The PCR-RFLP results showed that only L. tropica exists in this province. It is also determined that the positivity rate with PCR was higher (96%) than by microscopic examination (64%) in the diagnosis of ACL. Our results indicate that the PCR-RFLP method is more sensitive and specific for the detection and differentiation of agents of ACL in this area.
Department of Medical Biology, Faculty of Medicine, Harran University, Sanliurfa, Turkey.
Feridun
F
Akkafa
Fuat
F
Dilmec
Zuhre
Z
Alpua
D000293:Adolescent; D000328:Adult; D000818:Animals; D002675:Child, Preschool; D017931:DNA Primers / Q000235:genetics; D016054:DNA, Protozoan / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D015252:Deoxyribonucleases, Type II Site-Specific / Q000378:metabolism; D019353:Endemic Diseases; D005260:Female; D006801:Humans; D007223:Infant; D007891:Leishmania / Q000145:classification* / Q000166:cytology / Q000235:genetics / Q000302:isolation & purification*; D016773:Leishmaniasis, Cutaneous / Q000453:epidemiology* / Q000469:parasitology*; D008297:Male; D008853:Microscopy; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D012680:Sensitivity and Specificity; D014421:Turkey / Q000453:epidemiology
D017931:DNA Primers; D016054:DNA, Protozoan; D015252:Deoxyribonucleases, Type II Site-Specific; C024403:GGCC-specific type II deoxyribonucleases