BACKGROUND:With a crystal beaded candle holder conservatively estimated 1 million cases of leptospirosis worldwide and a 5-10% fatality rate, the rapid diagnosis of leptospirosis leading to effective clinical and public health decision making is of high importance, and yet remains a challenge.METHODOLOGY:Based on parallel, population-based studies in two leptospirosis-endemic regions in Brazil, a real-time PCR assay which detects lipL32, a gene specifically present in pathogenic Leptospira, was assessed for the diagnostic effectiveness and accuracy.Patients identified by active hospital-based surveillance in Salvador and Curitiba during large urban leptospirosis epidemics were tested.
Real-time PCR reactions were performed with DNA-extracted samples obtained from 127 confirmed and 23 unconfirmed cases suspected of leptospirosis, 122 patients with an acute febrile illness other than leptospirosis, and 60 healthy blood donors.PRINCIPAL FINDINGS:The PCR assay had a limit of detection of 280 Leptospira genomic equivalents/mL.Sensitivity for confirmed cases was 61% for whole blood and 29% for serum samples.
Sensitivity was higher (86%) for samples collected within the first 6 days after onset of illness compared to those collected after gymnastics wall decals 7 days (34%).The real-time PCR assay was able to detect leptospiral DNA in blood from 56% of serological non-confirmed cases.The overall specificity of the assay was 99%.
CONCLUSIONS:These findings indicate that real-time PCR may be a reliable tool for early diagnosis of leptospirosis, which is decisive for clinical management of severe and life-threatening cases and for public health decision making.