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Validation of an Immunodiagnostic Assay for Detection of 13 Streptococcus pneumoniae Serotype-Specific Polysaccharides in Human Urine

Validation of an Immunodiagnostic Assay for Detection of 13 Streptococcus pneumoniae... Validation of an Immunodiagnostic Assay for Detection of 13 Streptococcus pneumoniae Serotype-Specific Polysaccharides in Human Urine Michael W. Pride a , Susanne M. Huijts b , Kangjian Wu a , Victor Souza a , Sherry Passador a , Chunyan Tinder a , Esther Song a , Arik Elfassy a , Lisa McNeil a , Ronald Menton c , Roger French c , Janice Callahan d , Chris Webber e , William C. Gruber f , Marc J. M. Bonten b , g and Kathrin U. Jansen a a Vaccine Research East and Early Development, Pfizer Research, Pearl River, New York, New York, USA b Julius Center for Public Health and Primary Care, University Medical Center Utrecht, Utrecht, The Netherlands c Biotechnology Unit and Oncology Clinical Research Statistics, Pfizer Research, Cambridge, Massachusetts, USA, and Pearl River, New York, USA d Callahan Associates Inc., La Jolla, California, USA e Vaccine Clinical Research, Pfizer Research, Maidenhead, United Kingdom f Vaccine Clinical Research, Pfizer Research, Pearl River, New York, USA g Department of Medical Microbiology, University Medical Center Utrecht, Utrecht, The Netherlands ABSTRACT To improve the clinical diagnosis of pneumococcal infection in bacteremic and nonbacteremic community-acquired pneumonia (CAP), a Luminex technology-based multiplex u rinary a ntigen d etection (UAD) diagnostic assay was developed and validated. The UAD assay can simultaneously detect 13 different serotypes of Streptococcus pneumoniae by capturing serotype-specific S. pneumoniae polysaccharides (PnPSs) secreted in human urine. Assay specificity is achieved by capturing the polysaccharides with serotype-specific monoclonal antibodies (MAbs) on spectrally unique microspheres. Positivity for each serotype was based on positivity cutoff values calculated from a standard curve run on each assay plate together with positive- and negative-control urine samples. The assay is highly specific, since significant signals are detected only when each PnPS was paired with its homologous MAb-coated microspheres. Validation experiments demonstrated excellent accuracy and precision. The UAD assay and corresponding positivity cutoff values were clinically validated by assessing 776 urine specimens obtained from patients with X-ray-confirmed CAP. The UAD assay demonstrated 97% sensitivity and 100% specificity using samples obtained from patients with bacteremic, blood culture-positive CAP. Importantly, the UAD assay identified Streptococcus pneumoniae (13 serotypes) in a proportion of individuals with nonbacteremic CAP, a patient population for which the pneumococcal etiology of CAP was previously difficult to assess. Therefore, the UAD assay provides a specific, noninvasive, sensitive, and reproducible tool to support vaccine efficacy as well as epidemiological evaluation of pneumococcal disease, including CAP, in adults. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Clinical and Vaccine Immunology American Society For Microbiology

Validation of an Immunodiagnostic Assay for Detection of 13 Streptococcus pneumoniae Serotype-Specific Polysaccharides in Human Urine

Clinical and Vaccine Immunology , Volume 19 (8): 1131 – Aug 1, 2012

Abstract

Validation of an Immunodiagnostic Assay for Detection of 13 Streptococcus pneumoniae Serotype-Specific Polysaccharides in Human Urine Michael W. Pride a , Susanne M. Huijts b , Kangjian Wu a , Victor Souza a , Sherry Passador a , Chunyan Tinder a , Esther Song a , Arik Elfassy a , Lisa McNeil a , Ronald Menton c , Roger French c , Janice Callahan d , Chris Webber e , William C. Gruber f , Marc J. M. Bonten b , g and Kathrin U. Jansen a a Vaccine Research East and Early Development, Pfizer Research, Pearl River, New York, New York, USA b Julius Center for Public Health and Primary Care, University Medical Center Utrecht, Utrecht, The Netherlands c Biotechnology Unit and Oncology Clinical Research Statistics, Pfizer Research, Cambridge, Massachusetts, USA, and Pearl River, New York, USA d Callahan Associates Inc., La Jolla, California, USA e Vaccine Clinical Research, Pfizer Research, Maidenhead, United Kingdom f Vaccine Clinical Research, Pfizer Research, Pearl River, New York, USA g Department of Medical Microbiology, University Medical Center Utrecht, Utrecht, The Netherlands ABSTRACT To improve the clinical diagnosis of pneumococcal infection in bacteremic and nonbacteremic community-acquired pneumonia (CAP), a Luminex technology-based multiplex u rinary a ntigen d etection (UAD) diagnostic assay was developed and validated. The UAD assay can simultaneously detect 13 different serotypes of Streptococcus pneumoniae by capturing serotype-specific S. pneumoniae polysaccharides (PnPSs) secreted in human urine. Assay specificity is achieved by capturing the polysaccharides with serotype-specific monoclonal antibodies (MAbs) on spectrally unique microspheres. Positivity for each serotype was based on positivity cutoff values calculated from a standard curve run on each assay plate together with positive- and negative-control urine samples. The assay is highly specific, since significant signals are detected only when each PnPS was paired with its homologous MAb-coated microspheres. Validation experiments demonstrated excellent accuracy and precision. The UAD assay and corresponding positivity cutoff values were clinically validated by assessing 776 urine specimens obtained from patients with X-ray-confirmed CAP. The UAD assay demonstrated 97% sensitivity and 100% specificity using samples obtained from patients with bacteremic, blood culture-positive CAP. Importantly, the UAD assay identified Streptococcus pneumoniae (13 serotypes) in a proportion of individuals with nonbacteremic CAP, a patient population for which the pneumococcal etiology of CAP was previously difficult to assess. Therefore, the UAD assay provides a specific, noninvasive, sensitive, and reproducible tool to support vaccine efficacy as well as epidemiological evaluation of pneumococcal disease, including CAP, in adults.

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References (49)

Publisher
American Society For Microbiology
Copyright
Copyright © 2012 by the American society for Microbiology.
ISSN
1556-6811
eISSN
1556-679X
DOI
10.1128/CVI.00064-12
pmid
22675155
Publisher site
See Article on Publisher Site

Abstract

Validation of an Immunodiagnostic Assay for Detection of 13 Streptococcus pneumoniae Serotype-Specific Polysaccharides in Human Urine Michael W. Pride a , Susanne M. Huijts b , Kangjian Wu a , Victor Souza a , Sherry Passador a , Chunyan Tinder a , Esther Song a , Arik Elfassy a , Lisa McNeil a , Ronald Menton c , Roger French c , Janice Callahan d , Chris Webber e , William C. Gruber f , Marc J. M. Bonten b , g and Kathrin U. Jansen a a Vaccine Research East and Early Development, Pfizer Research, Pearl River, New York, New York, USA b Julius Center for Public Health and Primary Care, University Medical Center Utrecht, Utrecht, The Netherlands c Biotechnology Unit and Oncology Clinical Research Statistics, Pfizer Research, Cambridge, Massachusetts, USA, and Pearl River, New York, USA d Callahan Associates Inc., La Jolla, California, USA e Vaccine Clinical Research, Pfizer Research, Maidenhead, United Kingdom f Vaccine Clinical Research, Pfizer Research, Pearl River, New York, USA g Department of Medical Microbiology, University Medical Center Utrecht, Utrecht, The Netherlands ABSTRACT To improve the clinical diagnosis of pneumococcal infection in bacteremic and nonbacteremic community-acquired pneumonia (CAP), a Luminex technology-based multiplex u rinary a ntigen d etection (UAD) diagnostic assay was developed and validated. The UAD assay can simultaneously detect 13 different serotypes of Streptococcus pneumoniae by capturing serotype-specific S. pneumoniae polysaccharides (PnPSs) secreted in human urine. Assay specificity is achieved by capturing the polysaccharides with serotype-specific monoclonal antibodies (MAbs) on spectrally unique microspheres. Positivity for each serotype was based on positivity cutoff values calculated from a standard curve run on each assay plate together with positive- and negative-control urine samples. The assay is highly specific, since significant signals are detected only when each PnPS was paired with its homologous MAb-coated microspheres. Validation experiments demonstrated excellent accuracy and precision. The UAD assay and corresponding positivity cutoff values were clinically validated by assessing 776 urine specimens obtained from patients with X-ray-confirmed CAP. The UAD assay demonstrated 97% sensitivity and 100% specificity using samples obtained from patients with bacteremic, blood culture-positive CAP. Importantly, the UAD assay identified Streptococcus pneumoniae (13 serotypes) in a proportion of individuals with nonbacteremic CAP, a patient population for which the pneumococcal etiology of CAP was previously difficult to assess. Therefore, the UAD assay provides a specific, noninvasive, sensitive, and reproducible tool to support vaccine efficacy as well as epidemiological evaluation of pneumococcal disease, including CAP, in adults.

Journal

Clinical and Vaccine ImmunologyAmerican Society For Microbiology

Published: Aug 1, 2012

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