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.

[1]  V. Sharma,et al.  Semi-automated fluorogenic PCR assays (TaqMan) forrapid detection of Escherichia coli O157:H7 and other shiga toxigenic E. coli. , 1999, Molecular and cellular probes.

[2]  Arun K. Bhunia,et al.  Detection of Low Levels of Listeria monocytogenes Cells by Using a Fiber-Optic Immunosensor , 2004, Applied and Environmental Microbiology.

[3]  F. Ligler,et al.  Quantitating staphylococcal enterotoxin B in diverse media using a portable fiber-optic biosensor. , 1996, Analytical biochemistry.

[4]  F. Gunzer,et al.  Rapid Detection of Enterohemorrhagic Escherichia coliby Real-Time PCR with Fluorescent Hybridization Probes , 2001, Journal of Clinical Microbiology.

[5]  D. Lim,et al.  Detection of microorganisms and toxins with evanescent wave fiber-optic biosensors , 2003, Proc. IEEE.

[6]  X. Nou,et al.  Characterization of O157:H7 and other Escherichia coli isolates recovered from cattle hides, feces, and carcasses. , 2004, Journal of food protection.

[7]  P. Hatfield,et al.  Escherichia coli O157:H7 in microbial flora of sheep , 1996, Journal of clinical microbiology.

[8]  M. Doyle,et al.  An outbreak of Escherichia coli O157:H7 infections traced to jerky made from deer meat. , 1997 .

[9]  James C. Paton,et al.  Detection and Characterization of Shiga ToxigenicEscherichia coli by Using Multiplex PCR Assays forstx1, stx2,eaeA, Enterohemorrhagic E. coli hlyA,rfbO111, andrfbO157 , 1998, Journal of Clinical Microbiology.

[10]  H. Sheng,et al.  Rectal Administration of Escherichia coli O157:H7: Novel Model for Colonization of Ruminants , 2004, Applied and Environmental Microbiology.

[11]  S. Kern,et al.  Sodium boric acid: a Tris-free, cooler conductive medium for DNA electrophoresis. , 2004, BioTechniques.

[12]  D. Lane.  16S/23S rRNA sequencing , 1991 .

[13]  N. Pace,et al.  Differential amplification of rRNA genes by polymerase chain reaction , 1992, Applied and environmental microbiology.

[14]  K. E. Saxton,et al.  The Effect of Cattle Grazing on Indicator Bacteria in Runoff From a Pacific Northwest Watershed , 1982 .

[15]  G.P. Anderson,et al.  Development of an evanescent wave fiber optic biosensor , 1994, IEEE Engineering in Medicine and Biology Magazine.

[16]  Daniel V. Lim,et al.  DIRECT DETECTION OF ESCHERICHIA COLI 0157:H7 IN UNPASTEURIZED APPLE JUICE WITH AN EVANESCENT WAVE BIOSENSOR , 2001 .

[17]  A. Borczyk,et al.  Sequence heterogeneity of the eae gene and detection of verotoxin-producing Escherichia coli using serotype-specific primers , 1994, Epidemiology and Infection.

[18]  A. Bhunia,et al.  Analysis of environmental Escherichia coli isolates for virulence genes using the TaqMan® PCR system , 2003, Journal of applied microbiology.

[19]  D. Lim,et al.  Confirmation of viable E. coli O157:H7 by enrichment and PCR after rapid biosensor detection. , 2003, Journal of microbiological methods.

[20]  D A McCrae,et al.  Rapid detection of Escherichia coli O157:H7 in ground beef using a fiber-optic biosensor. , 1999, Journal of food protection.

[21]  D. Lim,et al.  A rapid and automated fiber optic-based biosensor assay for the detection of Salmonella in spent irrigation water used in the sprouting of sprout seeds. , 2004, Journal of food protection.

[22]  D. Demarco,et al.  Detection of Escherichia coli O157:H7 in 10- and 25-gram ground beef samples with an evanescent-wave biosensor with silica and polystyrene waveguides. , 2002, Journal of food protection.

[23]  J. Brown,et al.  Detection of Clostridium botulinum toxin A using a fiber optic-based biosensor. , 1992, Analytical biochemistry.

[24]  P. Mead,et al.  Escherichia coli O157:H7 , 1998, The Lancet.

[25]  S. Phatak,et al.  Persistence of enterohemorrhagic Escherichia coli O157:H7 in soil and on leaf lettuce and parsley grown in fields treated with contaminated manure composts or irrigation water. , 2004, Journal of food protection.

[26]  D. Myers,et al.  ESCHERICHIA COLI AND FECAL-COLBFORM BACTERIA AS INDICATORS OF RECREATIONAL WATER QUALITY , 1993 .

[27]  K P Flint,et al.  The long-term survival of Escherichia coli in river water. , 1987, The Journal of applied bacteriology.

[28]  George P. Anderson,et al.  Water quality monitoring using an automated portable fiber optic biosensor: RAPTOR , 2001, SPIE Optics East.

[29]  M. Louie,et al.  PCR Detection of Escherichia coli O157:H7 Directly from Stools: Evaluation of Commercial Extraction Methods for Purifying Fecal DNA , 2000, Journal of Clinical Microbiology.

[30]  D. Lim,et al.  A rapid detection method for Vaccinia virus, the surrogate for smallpox virus , 2004, Biosensors and Bioelectronics.

[31]  L. Slutsker,et al.  Escherichia coli O157:H7 gastroenteritis and the hemolytic uremic syndrome: an emerging infectious disease. , 1999, Annual review of medicine.

[32]  Frances S. Ligler,et al.  Fiber-Optic Biosensor for the Detection of Hazardous Materials , 1993 .

[33]  C. Schutt,et al.  The enhancement of PCR amplification by low molecular weight amides. , 2001, Nucleic acids research.

[34]  E. Stackebrandt,et al.  Nucleic acid techniques in bacterial systematics , 1991 .

[35]  D. Demarco,et al.  RECOVERY OF ESCHERICHIA COLI O157:H7 FROM FIBER OPTIC WAVEGUIDES USED FOR RAPID BIOSENSOR DETECTION , 2002 .