A nanoparticle amplification based quartz crystal microbalance DNA sensor for detection of Escherichia coli O157 : H7

Abstract

A quartz crystal microbalance (QCM) DNA sensor, based on the nanoparticle amplification method, was developed for detection of Escherichia coli O157:H7. A thiolated single-stranded DNA (ssDNA) probe specific to E. coli O157:H7 eaeA gene was immobilized onto the QCM sensor surface through self-assembly. The hybridization was induced by exposing the ssDNA probe to the complementary target DNA, and resulted in the mass change and therefore frequency change of the QCM. Streptavidin conjugated Fe3O4 nanoparticles (average diameter = 145 nm) were used as ``mass enhancers'' to amplify the frequency change. Synthesized biotinylated oligonucleotides as well as E. coli O157:H7 eaeA gene fragments (151 bases) amplified using asymmetric PCR with biotin labeled primers were tested. As low as 10-12 M synthesized oligonucleotides and 2.67 x 10(2) Colony forming unit (CFU)/ml E. coli 0157:H7 cells can be detected by the sensor. Linear correlation. between frequency change andlogarithmic numberof bacterial cell concentration was found for E. coliO157:H7 from 2.67 x 10(2) to 2.67 x 10(6) CFU/ml. (c) 2005 Elsevier B.V. All rights reserved.