Detection of PCR products of Escherichia coli O157:H7 in human stool samples using surface plasmon resonance (SPR).
A method for the rapid detection of verotoxin-producing Escherichia coli O157:H7 in stools was evaluated. Strains possessing Shiga toxin-2 (stx-2) genes were isolated from stool samples and amplified using oligonucleotide primers. Stools spiked with cultured E. coli O157:H7 (strain 298 or strain 1646) were detected to be polymerase chain reaction (PCR) positive at 10(2) cfu per 0.1 g of stool. Stool samples from patients and healthy carriers showed a high correlation between positive results for a PCR and the presence of verotoxin-producing E. coli O157:H7, confirmed by isolation of serotype O157:H7 on sorbitol MacConkey medium (10 of 10 stool samples). These PCR products could be detected using a BIAcore 2000 surface plasmon resonance device using peptide nucleic acid as a sensor probe. In this report we use this method for the rapid detection of DNA from significant pathogenic organisms.
Kai
E
E
Research Center for Advanced Science and Technology, The University of Tokyo, Komaba, Japan.
Ikebukuro
K
K
Hoshina
S
S
Watanabe
H
H
Karube
I
I
D002353:Carrier State / Q000382:microbiology; D004269:DNA, Bacterial / Q000032:analysis; D004927:Escherichia coli Infections / Q000382:microbiology; D019453:Escherichia coli O157 / Q000235:genetics / Q000302:isolation & purification*; D005243:Feces / Q000382:microbiology*; D006801:Humans; D020135:Peptide Nucleic Acids / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods*; D022641:Shiga Toxin 2 / Q000235:genetics* / Q000378:metabolism; D020349:Surface Plasmon Resonance / Q000379:methods*
D004269:DNA, Bacterial; D020135:Peptide Nucleic Acids; D022641:Shiga Toxin 2