Simultaneous Detection of Pathogenic and Saprophyte Leptospira in Human Plasma by Multiplex Taqman Real Time PCR

Abstract


Background: Leptospirosis is an infectious disease caused by pathogenic and saprophytic Leptospira species. The clinical and laboratory diagnosis of this infection is complicated. However, timely diagnosis of leptospirosis is essential for treatment of this disease. Conventional laboratory methods are incapable in the early diagnosis of it. Molecular tests such as real time PCR are very efficient when diagnosing it.

Objectives: In this study, we designed and developed a multiplex Taqman real time PCR to simultaneously detect saprophyte and pathologic Leptospira in clinical samples.

Methods: 250 human plasma samples were obtained from suspected patients. Two pair specific primers and the corresponding probe for detecting pathogenic and saprophytic Leptospira were designed and established in a single tube. The developed tests were run on all DNA extracted from the samples.

Results: Of the 250 samples, 93 (37.2%) were positive for pathogenic and 15 (6%) for non-pathogenic cases. In two samples, pathogenic and non-pathogenic DNA strains were simultaneously positive.

Conclusions: Based on our finds, the real time PCR is a suitable test for the diagnosis of leptospirosis and differentiation between pathogen and saprophyte Leptospira simultaneously.