Enrichment and detection of escherichia coli O157:H7 from water samples using an antibody modified microfluidic chip

Udara Dharmasiri, Małgorzata A. Witek, Andre A. Adams, John K. Osiri, Mateusz L. Hupert, Thomas S. Bianchi, Daniel L. Roelke, Steven A. Soper

Research output: Contribution to journalArticlepeer-review

95 Scopus citations

Abstract

Low abundant (< 100 cells mL-1) E. coli O157:H7 cells were isolated and enriched from environmental water samples using a microfluidic chip. The poly(methylmethacrylate), PMMA, chip contained 8 devices, each equipped with 16 curvilinear high aspect ratio channels that were covalently decorated with polyclonal anti-O157 antibodies (pAb) and could search for rare cells through a pAb mediated process. The chip could process independently 8 different samples or one sample using 8 different parallel inputs to increase volume processing throughput. After cell enrichment, cells were released and enumerated using benchtop real-time quantitative polymerase chain reaction (PCR), targeting genes which effectively discriminated the O157:H7 serotype from other nonpathogenic bacteria. The recovery of target cells from water samples was determined to be ∼72%, and the limit-of-detection was found to be 6 colony forming units (cfu) using the slt1 gene as a reporter. We subsequently performed analysis of lake and wastewater samples. The simplicity in manufacturing and ease of operation makes this device attractive for the selection of pathogenic species from a variety of water supplies suspected of containing bacterial pathogens at extremely low frequencies.

Original languageEnglish (US)
Pages (from-to)2844-2849
Number of pages6
JournalAnalytical Chemistry
Volume82
Issue number7
DOIs
StatePublished - Apr 1 2010
Externally publishedYes

ASJC Scopus subject areas

  • Analytical Chemistry

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