Detection of Leishmania major using PCR-ELISA

Abstract

Background and objectives: Cutaneous leishmaniasis is endemic in most areas of Iran, and the diagnosis of its species is essential for controlling the disease. Leishmania major is the causative agent for cutaneous leishmaniasis in humans. Molecular methods are generally more sensitive than microscopic methods. The present study aimed to use a polymerase chain reaction-enzyme linked immunosorbent assay (PCR-ELISA) technique for detecting live L. major from wounds of patients with cutaneous leishmaniasis. Methods: In the present study, a standard strain of L. major promastigotes was used as the positive control for purification of DNA. The Novy–MacNeal–Nicolle and RPMI-1640 media were used for reproduction of parasites. DNA was isolated from specimens taken from 35 patients with suspected cutaneous leishmaniasis whose disease was confirmed by direct smear method. The PCR-ELISA technique was later applied by using the standard strain, patient specimens, and primers specific for the 18s rRNA. Results: Out of 35 patients, 17 (48.6%) were male and 18 (51.4%) were female. In addition, 8.6% of the patients lived in the Gonbad-e Kavus County, while all patients had been infected in villages around Gonbad-e Kavus. Of 35 patients with confirmed cutaneous leishmaniasis according to the direct smear method, 31 patients (86.31%) had leishmaniasis based on the PCR method and the PCR-ELISA methods. Conclusion: Based on the results, the PCR-ELISA method is more sensitive and accurate for detecting L. major.

Keywords

• leishmania major
• pcr-elisa
• patients