Rapid, specific and sensitive electrochemical detection of foodborne bacteria.
Electrochemical biochips are an emerging tool for point-of-care diagnostic systems in medicine, food and environmental monitoring. In the current study, a thermostable reporter enzyme, esterase 2 (EST2) from Alicyclobacillus acidocaldarius, is used for specific and sensitive detection of bacteria by one-step rRNA/DNA hybridization between a bacterium-specific capture oligodeoxynucleotide (ODN), bacterial 16S rRNA and an uniform EST2-ODN reporter conjugate. The detection limit corresponds to approximately 500 colony forming units (cfu) Escherichia coli. Beside high sensitivity, the application of electrochemical biochips allows discrimination of two gram-negative and two gram-positive bacteria demonstrating the specificity and the potential for parallel detection of microorganisms. The feasibility of identification of foodborne bacteria was studied with meat juice contaminated with E. coli. This detection system has the capability to be applied for monitoring of bacterial food contamination.
Pöhlmann
Christopher
C
Laboratorium für Biochemie, Universität Bayreuth, Universitätsstrasse 30, 95440 Bayreuth, Germany.
Wang
Yiran
Y
Humenik
Martin
M
Heidenreich
Bernd
B
Gareis
Manfred
M
Sprinzl
Mathias
M
D001407:Bacillus / Q000201:enzymology / Q000235:genetics*; D001426:Bacterial Proteins / Q000138:chemical synthesis / Q000378:metabolism; D001483:Base Sequence; D015374:Biosensing Techniques / Q000191:economics / Q000295:instrumentation / Q000379:methods*; D055664:Electrochemical Techniques / Q000191:economics / Q000295:instrumentation / Q000379:methods*; D004950:Esterases / Q000138:chemical synthesis / Q000378:metabolism*; D006090:Gram-Negative Bacteria / Q000201:enzymology / Q000235:genetics; D006094:Gram-Positive Bacteria / Q000201:enzymology / Q000235:genetics; D008460:Meat / Q000032:analysis / Q000382:microbiology; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization / Q000379:methods; D009838:Oligodeoxyribonucleotides / Q000138:chemical synthesis / Q000235:genetics*; D012329:RNA, Bacterial / Q000032:analysis / Q000235:genetics / Q000302:isolation & purification; D012336:RNA, Ribosomal, 16S / Q000032:analysis* / Q000235:genetics / Q000302:isolation & purification; D012680:Sensitivity and Specificity; D013997:Time Factors
D001426:Bacterial Proteins; D009838:Oligodeoxyribonucleotides; D012329:RNA, Bacterial; D012336:RNA, Ribosomal, 16S; D004950:Esterases