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...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2005-12-01
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| Series: | Annals of Clinical Microbiology and Antimicrobials |
| Online Access: | http://www.ann-clinmicrob.com/content/4/1/22 |
| _version_ | 1828237688235360256 |
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| author | Kar Kalipada Ledizet Michel Li Li Koski Raymond A Kazmierczak Barbara I |
| author_facet | Kar Kalipada Ledizet Michel Li Li Koski Raymond A Kazmierczak Barbara I |
| author_sort | Kar Kalipada |
| collection | DOAJ |
| 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> |
| first_indexed | 2024-04-12T20:51:16Z |
| format | Article |
| id | doaj.art-2ed6955d3cd3451da19d686614e751a3 |
| institution | Directory Open Access Journal |
| issn | 1476-0711 |
| language | English |
| last_indexed | 2024-04-12T20:51:16Z |
| publishDate | 2005-12-01 |
| publisher | BMC |
| record_format | Article |
| series | Annals of Clinical Microbiology and Antimicrobials |
| spelling | doaj.art-2ed6955d3cd3451da19d686614e751a32022-12-22T03:17:06ZengBMCAnnals 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 |
| 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 |
| title | 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_short | 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 |
| url | http://www.ann-clinmicrob.com/content/4/1/22 |
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