An indirect enzyme-linked immunosorbent assay for rapid and quantitative assessment of Type III virulence phenotypes of <it>Pseudomonas aeruginosa </it>isolates

<p>Abstract</p> <p>Background</p> <p>The presence of a Type III secretion system in clinical isolates of <it>Pseudomonas aeruginosa </it>is associated with severe disease and poor outcomes in infections caused by this pathogen. We describe an indirect enzyme...

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Main Authors: Kar Kalipada, Ledizet Michel, Li Li, Koski Raymond A, Kazmierczak Barbara I
Format: Article
Language:English
Published: BMC 2005-12-01
Series:Annals of Clinical Microbiology and Antimicrobials
Online Access:http://www.ann-clinmicrob.com/content/4/1/22
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spelling doaj-2ed6955d3cd3451da19d686614e751a32020-11-25T00:56:47ZengBMCAnnals of Clinical Microbiology and Antimicrobials1476-07112005-12-01412210.1186/1476-0711-4-22An indirect enzyme-linked immunosorbent assay for rapid and quantitative assessment of Type III virulence phenotypes of <it>Pseudomonas aeruginosa </it>isolatesKar KalipadaLedizet MichelLi LiKoski Raymond AKazmierczak Barbara I<p>Abstract</p> <p>Background</p> <p>The presence of a Type III secretion system in clinical isolates of <it>Pseudomonas aeruginosa </it>is associated with severe disease and poor outcomes in infections caused by this pathogen. We describe an indirect enzyme-linked immunosorbent assay that rapidly and quantitatively detects two exotoxins, ExoU and ExoT, and two structural components, PopD and PcrV, of the <it>P. aeruginosa </it>Type III secretion system after in-vitro growth in a calcium-free minimal medium.</p> <p>Methods</p> <p>We used this assay to characterize the Type III secretion phenotype of 74 clinical isolates of <it>P. aeruginosa</it>. Findings were compared with results of standard immunoblotting and correlated with Type III secretion-dependent virulence of isolates toward cultured epithelial cells.</p> <p>Results</p> <p>Results of the ELISA assay were concordant with immunoblot detection of the secreted antigens for 73 of 74 isolates. The Type III secretion phenotype assessed by this immunoassay predicted bacterial virulence toward epithelial cells in vitro for all but five of the clinical isolates.</p> <p>Conclusion</p> <p>The availability of an ELISA assay for rapid detection of Type III secreted virulence factors will facilitate large clinical studies to examine whether the Type III secretion phenotype of a <it>P. aeruginosa </it>isolate predicts the course of clinical disease in a patient and should be taken into account in determining optimal treatment strategies for infected patients.</p> http://www.ann-clinmicrob.com/content/4/1/22
collection DOAJ
language English
format Article
sources DOAJ
author Kar Kalipada
Ledizet Michel
Li Li
Koski Raymond A
Kazmierczak Barbara I
spellingShingle Kar Kalipada
Ledizet Michel
Li Li
Koski Raymond A
Kazmierczak Barbara I
An indirect enzyme-linked immunosorbent assay for rapid and quantitative assessment of Type III virulence phenotypes of <it>Pseudomonas aeruginosa </it>isolates
Annals of Clinical Microbiology and Antimicrobials
author_facet Kar Kalipada
Ledizet Michel
Li Li
Koski Raymond A
Kazmierczak Barbara I
author_sort Kar Kalipada
title An indirect enzyme-linked immunosorbent assay for rapid and quantitative assessment of Type III virulence phenotypes of <it>Pseudomonas aeruginosa </it>isolates
title_short An indirect enzyme-linked immunosorbent assay for rapid and quantitative assessment of Type III virulence phenotypes of <it>Pseudomonas aeruginosa </it>isolates
title_full An indirect enzyme-linked immunosorbent assay for rapid and quantitative assessment of Type III virulence phenotypes of <it>Pseudomonas aeruginosa </it>isolates
title_fullStr An indirect enzyme-linked immunosorbent assay for rapid and quantitative assessment of Type III virulence phenotypes of <it>Pseudomonas aeruginosa </it>isolates
title_full_unstemmed An indirect enzyme-linked immunosorbent assay for rapid and quantitative assessment of Type III virulence phenotypes of <it>Pseudomonas aeruginosa </it>isolates
title_sort indirect enzyme-linked immunosorbent assay for rapid and quantitative assessment of type iii virulence phenotypes of <it>pseudomonas aeruginosa </it>isolates
publisher BMC
series Annals of Clinical Microbiology and Antimicrobials
issn 1476-0711
publishDate 2005-12-01
description <p>Abstract</p> <p>Background</p> <p>The presence of a Type III secretion system in clinical isolates of <it>Pseudomonas aeruginosa </it>is associated with severe disease and poor outcomes in infections caused by this pathogen. We describe an indirect enzyme-linked immunosorbent assay that rapidly and quantitatively detects two exotoxins, ExoU and ExoT, and two structural components, PopD and PcrV, of the <it>P. aeruginosa </it>Type III secretion system after in-vitro growth in a calcium-free minimal medium.</p> <p>Methods</p> <p>We used this assay to characterize the Type III secretion phenotype of 74 clinical isolates of <it>P. aeruginosa</it>. Findings were compared with results of standard immunoblotting and correlated with Type III secretion-dependent virulence of isolates toward cultured epithelial cells.</p> <p>Results</p> <p>Results of the ELISA assay were concordant with immunoblot detection of the secreted antigens for 73 of 74 isolates. The Type III secretion phenotype assessed by this immunoassay predicted bacterial virulence toward epithelial cells in vitro for all but five of the clinical isolates.</p> <p>Conclusion</p> <p>The availability of an ELISA assay for rapid detection of Type III secreted virulence factors will facilitate large clinical studies to examine whether the Type III secretion phenotype of a <it>P. aeruginosa </it>isolate predicts the course of clinical disease in a patient and should be taken into account in determining optimal treatment strategies for infected patients.</p>
url http://www.ann-clinmicrob.com/content/4/1/22
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