Development of an immunochromatographic test strip to detect pneumolysin by monoclonal antibody capture method

Autor: Somya Sephalika, Nirmal K. Mohakud, Ayman K. Johargy, Naif A. Jalal, Farkad Bantun, Aditya K. Panda, Raju K. Mandal, Shafiul Haque, Bikash Ranjan Sahu
Jazyk: angličtina
Rok vydání: 2024
Předmět:
Zdroj: Journal of King Saud University: Science, Vol 36, Iss 6, Pp 103213- (2024)
Druh dokumentu: article
ISSN: 1018-3647
DOI: 10.1016/j.jksus.2024.103213
Popis: Background: The currently available methods for the detection of Streptococcus pneumoniae are labour-intensive, and involve time-consuming steps needing skilled technicians to perform the assays. Therefore, we developed a rapid lateral flow based immunochromatographic test strip for detection of S. pneumoniae using pneumolysin using monoclonal antibody capture approach. Methodology − The strip was designed by assembling a sample pad, conjugation pad, nitrocellulose membrane and adsorbent pad supported by a plastic backing card. Optimization of control spot was done by using three different concentrations of monoclonal antibody (0.1 µg, 0.25 g and 0.5 µg). Monoclonal antibody coupled with gold nanoparticles was immobilized on conjugate pad. Recombinant pneumolysin at three different concentrations (10 ng/ml, 50 ng/ml and 100 ng/ml) was used. Diluted pneumolysin was poured onto the sample pad which migrated through the conjugate pad containing gold antibody conjugate and the antigen–antibody complex migrated to the test spot to bind to the polyclonal antibody. The remaining free antibody bound to the control spot to visualize a control spot. Results: Control spot was optimized at 0.5 µg of mouse IgG. Of three different concentrations of Pneumolysin, the spot was best visualized at 50 ng/ml Pneumolysin concentration. Conclusion: Our ICT based device could detect a commercially available recombinant Pneumolysin of S. pneumoniae. Future study is directed to validate the strip for detection of Pneumolysin using biological fluids of patients (particularly children) infected with S. pneumoniae.
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