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Strategies to improve the surface plasmon resonance-based immmunodetection of bacterial cells

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Abstract

We have made a comparison of (a) different surface chemistries of SPR sensor chips (such as carboxymethylated dextran and carboxymethylated C1) and (b) of different assay formats (direct, sandwich and subtractive immunoassay) in order to improve the sensitivity of the determination of the model bacteria Acidovorax avenae subsp. citrulli (Aac). The use of the carboxymethylated sensor chip C1 resulted in a better sensitivity than that of carboxymethylated dextran CM5 in all the assay formats. The direct assay format, in turn, exhibits the best sensitivity. Thus, the combination of a carboxymethylated sensor chip C1 with the direct assay format resulted in the highest sensitivity for Aac, with a limit of detection of 1.6 × 106 CFU mL-1. This SPR immunosensor was applied to the detection of Aac in watermelon leaf extracts spiked with the bacteria, and the lower LOD is 2.2 × 107 CFU mL−1.

Possible strategies to improve the surface plasmon resonance-based immmunodetection of bacterial cells Acidovorax avenae subsp. citrulli (Aac) was used as a model pathogen. Two different sensor surfaces (carboxymethylated dextran CM5 and carboxymethylated C1) were compared. Direct detection, sandwich system and subtractive assay were investigated. The combination of a C1 chip with the direct assay format resulted in the highest sensitivity for Aac, with a limit of detection of 1.6*106 CFU mL−1

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Acknowledgments

This project and a PhD scholarship to RC were supported by National Science and Technology Development Agency (NSTDA, Thailand). NK was partially supported by a Marie Curie Fellowship under a project entitled “PathFinder” (project number 910608). We are grateful to Prof. Dr. Morakot Tanticharoen for her mentorship on Biosensor Program at NSTDA.

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Authors and Affiliations

  1. National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency (NSTDA), 113 Thailand Science Park, Phahonyothin Road, Pathumthani, 12120, Thailand

    Ratthaphol Charlermroj, Oraprapai Gajanandana, Orawan Himananto & Nitsara Karoonuthaisiri

  2. Institute for Global Food Security, School of Biological Sciences, Queen’s University Belfast, David Keir Building, Stranmillis Road, Belfast, BT9 5AG, UK

    Ratthaphol Charlermroj, Michalina Oplatowska, Irene R. Grant, Nitsara Karoonuthaisiri & Christopher T. Elliott

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  1. Ratthaphol Charlermroj
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  2. Michalina Oplatowska
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  3. Oraprapai Gajanandana
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  4. Orawan Himananto
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  5. Irene R. Grant
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  6. Nitsara Karoonuthaisiri
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  7. Christopher T. Elliott
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Correspondence to Ratthaphol Charlermroj.

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Charlermroj, R., Oplatowska, M., Gajanandana, O. et al. Strategies to improve the surface plasmon resonance-based immmunodetection of bacterial cells. Microchim Acta 180, 643–650 (2013). https://doi.org/10.1007/s00604-013-0975-x

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  • DOI: https://doi.org/10.1007/s00604-013-0975-x

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