PMID- 15320634
OWN - NLM
STAT- MEDLINE
DCOM- 20040914
LR  - 20191210
IS  - 1387-2176 (Print)
IS  - 1387-2176 (Linking)
VI  - 6
IP  - 2
DP  - 2004 Jun
TI  - Development of a novel hand-held immunoassay for the detection of 
      enterohemorrhagic Escherichia coli O157:H7.
PG  - 125-30
AB  - Escherichia coli O157:H7 is an important human pathogen responsible for numerous 
      outbreaks of hemorrhagic colitis and hemolytic uremic syndrome world-wide. A 
      portable detection device is needed for field testing and point-of-care testing. 
      Poly(dimethylsiloxane) (PDMS) is a solid substrate well suited for adsorption of 
      macromolecules. The purpose of this study was to develop a hand-held 
      enzyme-linked immunosorbent assay (ELISA) for the detection of E. coli O157:H7 
      antigens. The prototype consisted of three modules: one for loading reagents, a 
      second for immunosensor detection, and a third for discharge of wastes. Reagent 
      delivery was achieved by using 1-ml syringes embedded within the loading module. 
      The detection module was based on a PDMS layer on which varying concentrations of 
      E. coli O157:H7 antigens, and negative controls (Lactobacillus rhamnosus R011 and 
      phosphate-buffered saline) were passively adsorbed. Commercially available goat 
      anti-E. coli O157:H7 antibody was used as a primary antibody, a donkey anti-goat 
      IgG conjugated with horseradish peroxidase was used as a secondary antibody, and 
      a precipitating substrate was employed for colorimetric detection. Results 
      obtained with the prototype were compared to those obtained using a conventional 
      nitrocellulose membrane-based immunoassay. Using the prototype, E. coli O157:H7 
      antigens, in quantities from 4 to 400 ng, were accurately detected. This 
      detection limit was comparable to that observed using conventional dot-blot 
      assays. The PDMS layer could be re-used without loss of sensitivity. Colorimetric 
      detection could be visualized readily without the need for sophisticated 
      equipment. Furthermore, this device can be adapted to perform diagnostics for 
      other microbial pathogens currently detected using immunoassay methodology.
FAU - Lin, Frank Y H
AU  - Lin FY
AD  - Research Institute, Hospital for Sick Children, University of Toronto, Toronto, 
      Ontario, Canada.
FAU - Sherman, Philip M
AU  - Sherman PM
FAU - Li, Dongqing
AU  - Li D
LA  - eng
PT  - Comparative Study
PT  - Evaluation Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Validation Study
PL  - United States
TA  - Biomed Microdevices
JT  - Biomedical microdevices
JID - 100887374
RN  - 0 (Dimethylpolysiloxanes)
RN  - 0 (Membranes, Artificial)
RN  - 0 (Silicones)
RN  - 63148-62-9 (baysilon)
SB  - IM
MH  - Colorimetry/*methods
MH  - *Dimethylpolysiloxanes
MH  - Environmental Monitoring/instrumentation/methods
MH  - Enzyme-Linked Immunosorbent Assay/*instrumentation/methods
MH  - Equipment Design
MH  - Equipment Failure Analysis
MH  - Escherichia coli O157/*isolation & purification
MH  - Immunoassay/*instrumentation/methods
MH  - Immunoblotting/instrumentation/methods
MH  - Membranes, Artificial
MH  - Miniaturization
MH  - Reproducibility of Results
MH  - Sensitivity and Specificity
MH  - *Silicones
EDAT- 2004/08/24 05:00
MHDA- 2004/09/15 05:00
CRDT- 2004/08/24 05:00
PHST- 2004/08/24 05:00 [pubmed]
PHST- 2004/09/15 05:00 [medline]
PHST- 2004/08/24 05:00 [entrez]
AID - 10.1023/b:bmmd.0000031749.02570.75 [doi]
PST - ppublish
SO  - Biomed Microdevices. 2004 Jun;6(2):125-30. doi: 
      10.1023/b:bmmd.0000031749.02570.75.