Detection of Leptospira interrogans in plasma and urine samples by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP)

摘要

Acute phase of leptospirosis has high mortality. A diagnosis can be designed with high sensitivity by testing blood and urine samples by polymerase chain reaction (PCR) for early diagnosis of this phase of disease. The aim of this research was to detect Leptospira interrogans by PCR technique in plasma and urine samples which were taken from symptomatic patients. Clinical isolates were obtained in hospitals of north Iran, and analyzed by PCR method and confirmed with restriction fragment length polymorphism (RFLP). DNA was extracted from plasma and urine samples and the quantity of L. interrogans DNA was determined by using PCR technique with LP1 and Lp2 primers. RFLP technique was used by Alu Ι restriction enzyme for confirming of detection of one of the prevalent serogroup of leptospira in Mazandaran Province. In this study, more than 90 blood and urine samples were examined, 10 (9%) blood and 3 (3%) urine samples became positive by molecular method. The high specificity and sensitivity of molecular assay provide valuable tools for the early diagnosis of acute leptospirosis.