| Abstract: | This study describes the development and application of a new PCRassay for the specific detection of pathogenic leptospires and its comparisonwith a previously reported PCR protocol. New primers were designed for PCRoptimization and evaluation in artificially-infected paraffin-embedded tissues.PCR was then applied to post-mortem, paraffin-embedded samples, followed byamplicon sequencing. The PCR was more efficient than the reported protocol,allowing the amplification of expected DNA fragment from the artificiallyinfected samples and from 44% of the post-mortem samples. The sequences ofPCR amplicons from different patients showed >99% homology withpathogenic leptospires DNA sequences. The applicability of a highly sensitiveand specific tool to screen histological specimens for the detection ofpathogenic Leptospira spp. would facilitate a better assessmentof the prevalence and epidemiology of leptospirosis, which constitutes a healthproblem in many countries. |
