| dc.creator | Waggoner, Jesse J | |
| dc.creator | Balassiano, Ilana | |
| dc.creator | Abeynayake, Janaki | |
| dc.creator | Sahoo, Malaya K | |
| dc.creator | Mohamed-Hadley, Alisha | |
| dc.creator | Liu, Yuanyuan | |
| dc.creator | Vita-Brasil, Juliana Magalhães | |
| dc.creator | Pinsky, Benjamin A | |
| dc.date | 2015-06-17T12:05:20Z | |
| dc.date | 2015-06-17T12:05:20Z | |
| dc.date | 2014 | |
| dc.date.accessioned | 2023-09-26T22:26:13Z | |
| dc.date.available | 2023-09-26T22:26:13Z | |
| dc.identifier | WAGGONER, Jesse J. et al. Sensitive Real-Time PCR Detection of Pathogenic Leptospira spp. and a Comparison of Nucleic Acid Amplification Methods for the Diagnosis of Leptospirosis. Plos One, v.9, n.11, 8p, 2014. | |
| dc.identifier | https://www.arca.fiocruz.br/handle/icict/10866 | |
| dc.identifier | 10.1371/journal.pone.0112356 | |
| dc.identifier.uri | https://repositorioslatinoamericanos.uchile.cl/handle/2250/8879191 | |
| dc.description | Background: Bacteria of the genus Leptospira, the causative agents of leptospirosis, are categorized into pathogenic and
non-pathogenic species. However, the benefit of using a clinical diagnostic that is specific for pathogenic species remains
unclear. In this study, we present the development of a real-time PCR (rtPCR) for the detection of pathogenic Leptospira (the
pathogenic rtPCR), and we perform a comparison of the pathogenic rtPCR with a published assay that detects all Leptospira
species [the undifferentiated febrile illness (UFI) assay] and a reference 16S Leptospira rtPCR, which was originally designed
to detect pathogenic species.
Methodology/Principal Findings: For the pathogenic rtPCR, a new hydrolysis probe was designed for use with primers
from the UFI assay, which targets the 16S gene. The pathogenic rtPCR detected Leptospira DNA in 37/37 cultured isolates
from 5 pathogenic and one intermediate species. Two strains of the non-pathogenic L. biflexa produced no signal. Clinical
samples from 65 patients with suspected leptospirosis were then tested using the pathogenic rtPCR and a reference
Leptospira 16S rtPCR. All 65 samples had tested positive for Leptospira using the UFI assay; 62 (95.4%) samples tested
positive using the pathogenic rtPCR (p = 0.24). Only 24 (36.9%) samples tested positive in the reference 16S rtPCR (p,0.0001
for comparison with the pathogenic rtPCR and UFI assays). Amplicon sequencing confirmed the detection of pathogenic
Leptospira species in 49/50 cases, including 3 cases that were only detected using the UFI assay.
Conclusions/Significance: The pathogenic rtPCR displayed similar sensitivity to the UFI assay when testing clinical
specimens with no difference in specificity. Both assays proved significantly more sensitive than a real-time molecular test
used for comparison. Future studies are needed to investigate the clinical and epidemiologic significance of more sensitive
Leptospira detection using these tests. | |
| dc.format | application/pdf | |
| dc.language | eng | |
| dc.publisher | Plos One | |
| dc.rights | open access | |
| dc.subject | Leptospirosis | |
| dc.subject | PCR | |
| dc.subject | Leptospira | |
| dc.subject | Leptospirose | |
| dc.subject | Reação em Cadeia da Polimerase | |
| dc.title | Sensitive Real-Time PCR Detection of Pathogenic Leptospira spp. and a Comparison of Nucleic Acid Amplification Methods for the Diagnosis of Leptospirosis | |
| dc.type | Article | |
