Please use this identifier to cite or link to this item: https://www.arca.fiocruz.br/handle/icict/11211
Title: Efficient Detection of Pathogenic Leptospires Using 16S Ribosomal RNA.
Authors: Backstedt, Brian T
Buyuktanir, Ozlem
Lindow, Janet C
Wunder Junior, Elsio Augusto
Reis, Mitermayer Galvão dos
Usmani-Brown, Sahar
Ledizet, Michel
Ko, Albert Icksang
Pal, Utpal
Affilliation: University of Maryland. College Park. Department of Veterinary Medicine and Virginia-Maryland Regional College of Veterinary Medicine. Maryland, USA
University of Maryland. College Park. Department of Veterinary Medicine and Virginia-Maryland Regional College of Veterinary Medicine. Maryland, USA
Yale University. School of Public Health. Department of Epidemiology of Microbial Diseases. New Haven, Connecticut, USA / Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, Brasil
Yale University. School of Public Health. Department of Epidemiology of Microbial Diseases. New Haven, Connecticut, USA
Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, Brasil
L2 Diagnostics. New Haven, Connecticut, USA
L2 Diagnostics. New Haven, Connecticut, USA
Yale University. School of Public Health. Department of Epidemiology of Microbial Diseases. New Haven, Connecticut, USA / Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, Brasil
University of Maryland. College Park. Department of Veterinary Medicine and Virginia-Maryland Regional College of Veterinary Medicine. Maryland, USA
Abstract: Pathogenic Leptospira species cause a prevalent yet neglected zoonotic disease with mild to life-threatening complications in a variety of susceptible animals and humans. Diagnosis of leptospirosis, which primarily relies on antiquated serotyping methods, is particularly challenging due to presentation of non-specific symptoms shared by other febrile illnesses, often leading to misdiagnosis. Initiation of antimicrobial therapy during early infection to prevent more serious complications of disseminated infection is often not performed because of a lack of efficient diagnostic tests. Here we report that specific regions of leptospiral 16S ribosomal RNA molecules constitute a novel and efficient diagnostic target for PCR-based detection of pathogenic Leptospira serovars. Our diagnostic test using spiked human blood was at least 100-fold more sensitive than corresponding leptospiral DNA-based quantitative PCR assays, targeting the same 16S nucleotide sequence in the RNA and DNA molecules. The sensitivity and specificity of our RNA assay against laboratory-confirmed human leptospirosis clinical samples were 64% and 100%, respectively, which was superior then an established parallel DNA detection assay. Remarkably, we discovered that 16S transcripts remain appreciably stable ex vivo, including untreated and stored human blood samples, further highlighting their use for clinical detection of L. interrogans. Together, these studies underscore a novel utility of RNA targets, specifically 16S rRNA, for development of PCR-based modalities for diagnosis of human leptospirosis, and also may serve as paradigm for detection of additional bacterial pathogens for which early diagnosis is warranted.
keywords: Leptospirose
Leptospira
Sorotipagem
Humanos
RNA bacteriano
Antígenos bacterianos
Issue Date: 2015
Publisher: Public Library of Science
Citation: BACKSTEDT, B. T. et al. Efficient Detection of Pathogenic Leptospires Using 16S Ribosomal RNA. PLoS One, v. 10, n. 6, p. e0128913, 2015.
DOI: 10.1371/journal.pone.0128913
ISSN: 1932-6203
Copyright: open access
Appears in Collections:BA - IGM - Artigos de Periódicos

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