Joseph M. Antenello,
Author Of 1 Presentation
HIGH THROUGHPUT MULTIPLEX URINE ANTIGEN DETECTION ASSAY FOR THE QUANTITATION OF SEROTYPE SPECIFIC PNEUMOCOCCAL POLYSACCHARIDES TO SUPPORT PNEUMOCOCCAL CONJUGATE VACCINE (PCV) PROGRAM (ID 1240)
- Gowrisankar Rajam, United States of America
- Rebecca J. Grant-Klein,
- Reshma Panemangalore,
- Lauren Ciminera,
- Stephanie Cooper,
- Roshni Patel,
- Katrina Nolan, United States of America
- Yuhua Zhang,
- Jennifer Nguyen, United States of America
- Thomas Steinmetz, United States of America
- Joseph M. Antenello,
- Leonard J. Rubinstein,
- Rocio Murphy,
Abstract
Background
Streptococcus pneumoniae is the most common cause of community acquired pneumonia despite robust vaccinations. With ~100 serotypes (ST) in circulation, surveillance is crucial to estimate the ST specific disease burden and assess vaccine efficacy/coverage. To address these needs, we have developed a high throughput multiplex Pneumococcal Urine Antigen Detection (PnUAD) assay for the quantitation of 15 serotype specific polysaccharides (Ps) in human urine.
Methods
PnUAD was qualified based on the assay precision, ruggedness, specificity, accuracy, limit of detection, selectivity and dilutional linearity. Test samples were individual or pooled normal human urine spiked with full-length vaccine grade polysaccharides or fragmented polysaccharides (<150 kDa).
Results
PnUAD was determined to be serotype specific, precise (<15% RSD), accurate (80%-127% recovery throughout the quantifiable range), and dilutable (<2-fold dilution bias per 10-fold dilution) for each of the 15 Pn serotypes. The PnUAD was also determined to be highly sensitive (LLOQ ranging between 0.0020 – 0.0781 ng/mL across the 15 serotypes), and selective (consistent PnPs recovery for each ST when spiked into different pre-dilutions of urine).
Conclusions
PnUAD met all performance expectations, and is considered qualified to detect serotype specific PnPs in human urine in subjects from epidemiology studies or vaccine clinical trials.