Use of rpoB gene analysis for detection and identification of leptospira species by direct sequencing

Abstract

Leptospirosis is an emerging disease for which culture and identification are partly unresolved. In fact, 16S rRNA-based sequencing is the most widely used PCR methodology that can detect such uncultivable pathogens. However, this assay has some limitations linked to potential problems of contamination, which hampers diagnosis. To overcome this, we have a simple PCR strategy involving targeting of the gene encoding the RNA polymerase β subunit (rpoB), a highly conserved enzyme. The sequence of the Leptospira rpoB gene was determined and compared with the published sequence. Our findings have significant implications for the development of a new tool for the identification of spirochetes, especially if clinical samples are contaminated or when the infecting strain is uncultivable. The consistent use of PCR has improved the early diagnosis of Leptospirosis but the limitation is that it cannot provide information on the infecting Leptospira strain which provides important epidemiological data.