title: High sensitive PCR method for detection of pathogenic Leptospira spp. in paraffin-embedded tissues.
abstract: This study describes the development and application of a new PCR assay for the specific detection of pathogenic leptospires and its comparison with a previously reported PCR protocol. New primers were designed for PCR optimization and evaluation in artificially-infected paraffin-embedded tissues. PCR was then applied to post-mortem, paraffin-embedded samples, followed by amplicon sequencing. The PCR was more efficient than the reported protocol, allowing the amplification of expected DNA fragment from the artificially infected samples and from 44% of the post-mortem samples. The sequences of PCR amplicons from different patients showed >99% homology with pathogenic leptospires DNA sequences. The applicability of a highly sensitive and specific tool to screen histological specimens for the detection of pathogenic Leptospira spp. would facilitate a better assessment of the prevalence and epidemiology of leptospirosis, which constitutes a health problem in many countries.
authors.affiliation: Department of Bacteriology-Mycology, Institute of Tropical Medicine ?Pedro Kourí?, Havana, Cuba.
authors.forename: Angel Alberto
authors.initials: AA
authors.lastname: Noda
authors.affiliation: Department of Bacteriology-Mycology, Institute of Tropical Medicine ?Pedro Kourí?, Havana, Cuba.
authors.forename: Islay
authors.initials: I
authors.lastname: Rodríguez
authors.affiliation: Department of Bacteriology-Mycology, Institute of Tropical Medicine ?Pedro Kourí?, Havana, Cuba.
authors.forename: Yaindrys
authors.initials: Y
authors.lastname: Rodríguez
authors.affiliation: Department of Pathology, Institute of Tropical Medicine ?Pedro Kourí?, Havana, Cuba.
authors.forename: Anamays
authors.initials: A
authors.lastname: Govín
authors.affiliation: Department of Bacteriology-Mycology, Institute of Tropical Medicine ?Pedro Kourí?, Havana, Cuba.
authors.forename: Carmen
authors.initials: C
authors.lastname: Fernández
authors.affiliation: Department of Bacteriology-Mycology, Institute of Tropical Medicine ?Pedro Kourí?, Havana, Cuba.
authors.forename: Ana Margarita
authors.initials: AM
authors.lastname: Obregón
mesh_terms: D017931:DNA Primers / Q000235:genetics*; D004269:DNA, Bacterial / Q000235:genetics*; D006801:Humans; D007919:Leptospira / Q000235:genetics*; D007922:Leptospirosis / Q000175:diagnosis*; D016612:Paraffin Embedding; D016133:Polymerase Chain Reaction; D012720:Severity of Illness Index; D016707:Tissue Fixation
chemical_list: D017931:DNA Primers; D004269:DNA, Bacterial