Rapid PCR confirmation of E. coli O157:H7 after evanescent wave fiber optic biosensor detection.
Escherichia coli O157:H7 is an enteric pathogen of public health importance, which is monitored by several government agencies. Many rapid detection tests have been developed to identify foodstuff and water supplies contaminated by E. coli O157:H7. However, these methods can be time consuming (24-48 h) due to the need to culture the bacteria to confirm detection results. Fiber optic biosensors can rapidly detect pathogens from complex matrices, yet confirmation tests can take up to 10h to complete. In addition, fiber optic biosensors can also be used to reduce the impact of PCR inhibitors present in complex matrices such as food and water. This paper presents methodologies to reduce the time necessary for confirmation from 10 to about 2 h, by direct PCR of bacteria from the fiber optic waveguides without the need for culture or enrichment steps.
Simpson
Joyce M
JM
University of South Florida, Department of Biology, Center for Biological Defense, 4202 E. Fowler Ave. SCA110, Tampa, 33620-5200, USA. jsimpson@cas.usf.edu
Lim
Daniel V
DV
D000907:Antibodies, Bacterial / Q000032:analysis* / Q000276:immunology; D015374:Biosensing Techniques / Q000295:instrumentation* / Q000379:methods; D002469:Cell Separation / Q000295:instrumentation / Q000379:methods; D020099:Coated Materials, Biocompatible / Q000737:chemistry; D015169:Colony Count, Microbial / Q000295:instrumentation* / Q000379:methods; D003199:Computer Systems; D004867:Equipment Design; D019544:Equipment Failure Analysis; D019453:Escherichia coli O157 / Q000145:classification / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification*; D005336:Fiber Optic Technology / Q000295:instrumentation* / Q000379:methods; D007118:Immunoassay / Q000295:instrumentation / Q000379:methods; D055100:Optical Fibers; D010783:Photometry / Q000295:instrumentation / Q000379:methods; D016133:Polymerase Chain Reaction / Q000295:instrumentation* / Q000379:methods; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D018511:Systems Integration
D000907:Antibodies, Bacterial; D020099:Coated Materials, Biocompatible