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Miniaturized 96-well ELISA chips for staphylococcal enterotoxin B detection using portable colorimetric detector

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Abstract

A previously developed fluorescence sensing platform, combining spatial illumination using electroluminescence (EL) semiconductor strips with charge coupled device (CCD)-based detection (EL-CCD), was adapted to a new 96-well chip for colorimetric immunological assays, enhancing the capabilities of the EL-CCD platform. The modified system was demonstrated using a colorimetric-based enzyme linked immunosorbent assay (ELISA) for detection of staphylococcal enterotoxin B (SEB). Limits of detection (LODs) of 3.9 ng/mL (±2.4 ng/mL) SEB were determined with the ELISA chip measured using the EL-CCD platform, following a standard 4-h ELISA protocol. The LODs were comparable to those obtained using standard 96-well ELISA plates measured using a standard laboratory 96-well plate reader. The miniature 96-well ELISA chip however required as little as 5-µL samples, representing a tenfold reduction in sample volume compared to a standard 96-well ELISA plates. The ELISA chip also demonstrated detection of SEB spiked into various food matrices (milk, mushrooms, and mayonnaise) using limited-to-no sample preparation, with LODs ranging from 3.9 to 18.5 ng/mL depending on the matrix. The EL-CCD platform is versatile, capable of multi-mode detection (e.g., fluorescent and colorimetric along with solution and solid phase assays), and could readily be applied to other field portable or point-of-care applications.

Detection of SEB using miniature ELISA chips coupled with a portable electroluminiscent-charge couple device (EL-CCD) detection system.

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Acknowledgments

This work was supported in part by the Office of Public Health emergency Preparedness (OPHEP) IAG 224-05-655 (to A. Rasooly and by FDA contract) and HHSF223200610765P (to Dr. Yordan Kostov) and CDRH/OSEL/Division of Biology funds. The mention of commercial products, their sources, or their use in connection with material reported herein is not to be construed as either an actual or implied endorsement of such products by the Department of Health and Human Services

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Authors and Affiliations

  1. Division of Biology, Office of Science and Engineering Laboratories, CDRH, FDA, Silver Spring, MD, 20993, USA

    Kim E. Sapsford, Jesse Francis, Steven Sun & Avraham Rasooly

  2. Center for Advanced Sensor Technology, University of Maryland Baltimore County (UMBC), Baltimore, MD, 21227, USA

    Jesse Francis, Steven Sun & Yordan Kostov

  3. National Cancer Institute (NIH/NCI), 6130 Executive Blvd. EPN, Room 6035A, Rockville, MD, 20892, USA

    Avraham Rasooly

Authors
  1. Kim E. Sapsford
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  2. Jesse Francis
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  3. Steven Sun
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  4. Yordan Kostov
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  5. Avraham Rasooly
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Corresponding author

Correspondence to Avraham Rasooly.

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Sapsford, K.E., Francis, J., Sun, S. et al. Miniaturized 96-well ELISA chips for staphylococcal enterotoxin B detection using portable colorimetric detector. Anal Bioanal Chem 394, 499–505 (2009). https://doi.org/10.1007/s00216-009-2730-z

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  • DOI: https://doi.org/10.1007/s00216-009-2730-z

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