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Multiplexed paper test strip for quantitative bacterial detection

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Abstract

Rapid, sensitive, on-site detection of bacteria without a need for sophisticated equipment or skilled personnel is extremely important in clinical settings and rapid response scenarios, as well as in resource-limited settings. Here, we report a novel approach for selective and ultra-sensitive multiplexed detection of Escherichia coli (non-pathogenic or pathogenic) using a lab-on-paper test strip (bioactive paper) based on intracellular enzyme (β-galactosidase (B-GAL) or β-glucuronidase (GUS)) activity. The test strip is composed of a paper support (0.5 × 8 cm), onto which either 5-bromo-4-chloro-3-indolyl-β-d-glucuronide sodium salt (XG), chlorophenol red β-galactopyranoside (CPRG) or both and FeCl3 were entrapped using sol–gel-derived silica inks in different zones via an ink-jet printing technique. The sample was lysed and assayed via lateral flow through the FeCl3 zone to the substrate area to initiate rapid enzyme hydrolysis of the substrate, causing a change from colorless-to-blue (XG hydrolyzed by GUS, indication of nonpathogenic E. coli) and/or yellow to red-magenta (CPRG hydrolyzed by B-GAL, indication of total coliforms). Using immunomagnetic nanoparticles for selective preconcentration, the limit of detection was ~5 colony-forming units (cfu) per milliliter for E. coli O157:H7 and ~20 cfu/mL for E. coli BL21, within 30 min without cell culturing. Thus, these paper test strips could be suitable for detection of viable total coliforms and pathogens in bathing water samples. Moreover, inclusion of a culturing step allows detection of less than 1 cfu in 100 mL within 8 h, making the paper tests strips relevant for detection of multiple pathogens and total coliform bacteria in beverage and food samples.

Pathogen Sensing Paper: Paper strips with ink-jet printed sensing zones can detect low levels of pathogenic or non-pathogenic bacteria. Incorporation of an immunomagnetic separation step results in selective detection of ~25 cfu of H7:O157 bacteria in under 1 h.

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Abbreviations

B-GAL:

β-galactosidase

CI:

Color intensity

cfu:

Colony-forming units

CPRG:

Chlorophenol red β-galactopyranoside

E. coli :

Escherichia coli

GUS:

β-glucuronidase

HB:

Hydrophobic barrier

IMS:

Immunomagnetic separation

LOD:

Limit of detection

MB-Ab:

Antibody-derivatized magnetic beads

PVAm:

Polyvinylamine

XG:

5-Bromo-4-chloro-3-indolyl-β-d-glucuronide sodium salt

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Acknowledgments

The authors thank the Natural Sciences and Engineering Research Council of Canada and the Sentinel-Canadian Network for the Development and Use of Bioactive Paper for funding. The authors also thank the Canada Foundation for Innovation and the Ontario Ministry of Research and Innovation for support of this work. The authors thank Puneet Chavan for his contribution to experimental data. YL holds the Canada Research Chair in Nucleic Acids Chemistry. JDB holds the Canada Research Chair in Bioanalytical Chemistry and Biointerfaces.

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Authors and Affiliations

  1. Department of Chemistry and Chemical Biology, McMaster University, 1280 Main St. West, Hamilton, ON, L8S 4M1, Canada

    S. M. Zakir Hossain, Cory Ozimok, Clémence Sicard & John D. Brennan

  2. Department of Biochemistry and Biomedical Sciences, McMaster University, 1200 Main St. West, Hamilton, ON, L8N 3Z5, Canada

    Sergio D. Aguirre, M. Monsur Ali & Yingfu Li

Authors
  1. S. M. Zakir Hossain
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  2. Cory Ozimok
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  3. Clémence Sicard
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  4. Sergio D. Aguirre
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  5. M. Monsur Ali
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  6. Yingfu Li
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  7. John D. Brennan
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Correspondence to John D. Brennan.

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Hossain, S.M.Z., Ozimok, C., Sicard, C. et al. Multiplexed paper test strip for quantitative bacterial detection. Anal Bioanal Chem 403, 1567–1576 (2012). https://doi.org/10.1007/s00216-012-5975-x

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  • DOI: https://doi.org/10.1007/s00216-012-5975-x

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