Detection of pathogenic Leptospira spp. through TaqMan polymerase chain reaction targeting the LipL32 gene

Abstract Rapid diagnosis of leptospirosis, through culture and/or serology, can be difficult without proper expertise and is often delayed because of the length of time required to obtain results. In this study, we developed a real-time polymerase chain reaction (PCR) assay using a TaqMan probe targ...

Full description

Saved in:
Bibliographic Details
Published in:Diagnostic microbiology and infectious disease 2009-07, Vol.64 (3), p.247-255
Main Authors: Stoddard, Robyn A, Gee, Jay E, Wilkins, Patricia P, McCaustland, Karen, Hoffmaster, Alex R
Format: Article
Language:English
Subjects:
Bacterial Outer Membrane Proteins - genetics
Bacteriology
Biological and medical sciences
Blood - microbiology
Diagnosis
Fundamental and applied biological sciences. Psychology
Humans
Infectious Disease
Infectious diseases
Internal Medicine
Leptospira
Leptospira - genetics
Leptospira - isolation & purification
Leptospira interrogans
Leptospirosis
Leptospirosis - diagnosis
LipL32
Lipoproteins - genetics
Medical sciences
Microbiology
Miscellaneous
Plasma - microbiology
Polymerase Chain Reaction - methods
Real-time PCR
Sensitivity and Specificity
Serum - microbiology
TaqMan
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Abstract Rapid diagnosis of leptospirosis, through culture and/or serology, can be difficult without proper expertise and is often delayed because of the length of time required to obtain results. In this study, we developed a real-time polymerase chain reaction (PCR) assay using a TaqMan probe targeting lipL32 , which is present only in pathogenic Leptospira spp. Using Leptospira interrogans serovar Icterohaemorrhagiae DNA, the lower limit of detection was found to be 20 genomic equivalents/reaction with a 95% cutoff value. The assay detected pathogenic Leptospira strains, but not intermediately pathogenic or nonpathogenic strains. When testing the assay on spiked clinical specimens, whole blood and plasma were better specimens for detecting the same initial number of leptospires compared with serum from clotted and centrifuged blood. Leptospira spiked at the same concentration was better detected in centrifuged urine. This real-time PCR assay with high specificity and sensitivity may prove to be a rapid method for diagnosing acute leptospirosis.
ISSN:0732-8893
1879-0070
DOI:10.1016/j.diagmicrobio.2009.03.014