Analytical and Bioanalytical Chemistry

, Volume 403, Issue 6, pp 1567–1576 | Cite as

Multiplexed paper test strip for quantitative bacterial detection

  • S. M. Zakir Hossain
  • Cory Ozimok
  • Clémence Sicard
  • Sergio D. Aguirre
  • M. Monsur Ali
  • Yingfu Li
  • John D. Brennan
Original Paper

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.

Figure

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.

Keywords

Bacteria detection Bioactive paper sensor Colorimetric 

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

Supplementary material

216_2012_5975_MOESM1_ESM.pdf (210 kb)
ESM 1(PDF 209 kb)

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Copyright information

© Springer-Verlag 2012

Authors and Affiliations

  • S. M. Zakir Hossain
    • 1
  • Cory Ozimok
    • 1
  • Clémence Sicard
    • 1
  • Sergio D. Aguirre
    • 2
  • M. Monsur Ali
    • 2
  • Yingfu Li
    • 2
  • John D. Brennan
    • 1
  1. 1.Department of Chemistry and Chemical BiologyMcMaster UniversityHamiltonCanada
  2. 2.Department of Biochemistry and Biomedical SciencesMcMaster UniversityHamiltonCanada

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