Application of a Multiplex PCR Assay for Molecular Identification of Pathogenic and Non-Pathogenic Leptospires based on lipL۳۲ and ۱۶S rRNA Genes

Leptospirosis is a serious zoonotic infection and the most prevalence disease is in the tropical and subtropical region. The definitive diagnosis of Leptospirosis, caused by spirochetes of the genus Leptospira infection is already using culture methods, serological tests such as the microscopic agglutination test (MAT) and molecular detection methods (PCR) are possible.  In this study, we used multiplex PCR method for detection of pathogenic and non - pathogenic Leptospira based on lipL۳۲ and ۱۶S rRNA genes. All serovars were obtained from the Leptospira Reference Laboratory of Microbiology Department, Razi Vaccine and Serum Research Institute, Karaj, Iran. The PCR product for the lipL۳۲ and ۱۶S rRNA genes was ۲۷۲ bp and ۲۴۰ bp respectively. The sensitivity amplification for the multiplex assay was ۱۰-۶ pg / μl for ۱۶S rRNA gene and ۱۰-۴ pg / μl for lipL۳۲ gene. The sensitivity for multiplex PCR was ۱۰-۳ pg / μl. The results supported the idea that multiplex PCR can be used to detect Leptospira samples. This method was also able to differentiate between saprophytic and pathogenic leptospires and was able to do so much easily than conventional methodologies. Due to the slow growth of Leptospira and the importance of time in diagnosis, molecular methods such as PCR are suggested.

کلیدواژه ها:

Leptospira ، ۱۶S rRNA gene ، lipL۳۲ gene ، Multiplex PCR ، molecular identification