Rapid, actionable diagnosis of urban epidemic leptospirosis using a pathogenic Leptospira lipL32-based real-time PCR assay.

Autor: Riediger IN; Molecular Biology Section, Central Laboratory of the State of Paraná, Curitiba, Paraná Brazil., Stoddard RA; National Center for Emerging and Zoonotic Infectious Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia, United States of America., Ribeiro GS; Gonçalo Moniz Institute, Oswaldo Cruz Foundation, Brazilian Ministry of Health, Salvador, Bahia, Brazil.; Institute of Collective Health, Federal University of the State of Bahia, Salvador, Bahia, Brazil., Nakatani SM; Molecular Biology Section, Central Laboratory of the State of Paraná, Curitiba, Paraná Brazil., Moreira SDR; Hospital das Clínicas, Federal University of the State of Paraná, Curitiba, Paraná, Brazil., Skraba I; Molecular Biology Section, Central Laboratory of the State of Paraná, Curitiba, Paraná Brazil., Biondo AW; Department of Veterinary Medicine, Federal University of the State of Paraná, Curitiba, Paraná, Brazil., Reis MG; Gonçalo Moniz Institute, Oswaldo Cruz Foundation, Brazilian Ministry of Health, Salvador, Bahia, Brazil., Hoffmaster AR; National Center for Emerging and Zoonotic Infectious Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia, United States of America., Vinetz JM; Division of Infectious Diseases, Department of Medicine, University of California San Diego School of Medicine, La Jolla, California, United States of America., Ko AI; Gonçalo Moniz Institute, Oswaldo Cruz Foundation, Brazilian Ministry of Health, Salvador, Bahia, Brazil.; Department of Epidemiology of Microbial Diseases, Yale School of Public Health, New Haven, Connecticut, United States of America., Wunder EA Jr; Gonçalo Moniz Institute, Oswaldo Cruz Foundation, Brazilian Ministry of Health, Salvador, Bahia, Brazil.; Department of Epidemiology of Microbial Diseases, Yale School of Public Health, New Haven, Connecticut, United States of America.
Jazyk: angličtina
Zdroj: PLoS neglected tropical diseases [PLoS Negl Trop Dis] 2017 Sep 15; Vol. 11 (9), pp. e0005940. Date of Electronic Publication: 2017 Sep 15 (Print Publication: 2017).
DOI: 10.1371/journal.pntd.0005940
Abstrakt: Background: With a 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 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.
Databáze: MEDLINE
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