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Direct detection and differentiation of pathogenic Leptospira species using a multi-gene targeted real time PCR approach.

Authors :
Ana Sofia Ferreira
Pedro Costa
Teresa Rocha
Ana Amaro
Maria Luísa Vieira
Ahmed Ahmed
Gertrude Thompson
Rudy A Hartskeerl
João Inácio
Source :
PLoS ONE, Vol 9, Iss 11, p e112312 (2014)
Publication Year :
2014
Publisher :
Public Library of Science (PLoS), 2014.

Abstract

Leptospirosis is a growing public and veterinary health concern caused by pathogenic species of Leptospira. Rapid and reliable laboratory tests for the direct detection of leptospiral infections in animals are in high demand not only to improve diagnosis but also for understanding the epidemiology of the disease. In this work we describe a novel and simple TaqMan-based multi-gene targeted real-time PCR approach able to detect and differentiate Leptospira interrogans, L. kirschneri, L. borgpeteresenii and L. noguchii, which constitute the veterinary most relevant pathogenic species of Leptospira. The method uses sets of species-specific probes, and respective flanking primers, designed from ompL1 and secY gene sequences. To monitor the presence of inhibitors, a duplex amplification assay targeting both the mammal β-actin and the leptospiral lipL32 genes was implemented. The analytical sensitivity of all primer and probe sets was estimated to be

Subjects

Subjects :
Medicine
Science

Details

Language :
English
ISSN :
19326203
Volume :
9
Issue :
11
Database :
Directory of Open Access Journals
Journal :
PLoS ONE
Publication Type :
Academic Journal
Accession number :
edsdoj.6cfd2808b8194de8a02290b7484a3f95
Document Type :
article
Full Text :
https://doi.org/10.1371/journal.pone.0112312