Detection of bacterial 16S rRNA using multivalent dendrimer-reporter enzyme conjugates.
Novel enzyme-oligodeoxynucleotide conjugate was synthesized to improve sensitivity of Escherichia coli 16S rRNA detection on gold electrodes. Thermostable esterase 2 from Alicyclobacillus acidocaldarius was multiply conjugated to a polyamidoamine dendrimer functionalized by one universal detector oligodeoxynucleotide. Three components rRNA/DNA hybridization between capture oligodeoxynucleotide covalently immobilized on a gold electrode, 16S rRNA and the multivalent esterase-dendrimer cluster was used for detection of E. coli. The linear dependence of the electrochemical signals to analyte concentration revealed a detection limit of 50 colony forming units E. coli, which represents a tenfold signal enhancement if compared to the detection limit achieved with monovalent esterase-oligodeoxynucleotide conjugate.
Pöhlmann
Christopher
C
Laboratorium für Biochemie, Universität Bayreuth, Bayreuth, Germany.
Humenik
Martin
M
Sprinzl
Mathias
M
D015374:Biosensing Techniques / Q000295:instrumentation*; D015169:Colony Count, Microbial / Q000295:instrumentation; D050091:Dendrimers / Q000737:chemistry; D004563:Electrochemistry / Q000295:instrumentation*; D004566:Electrodes; D004867:Equipment Design; D019544:Equipment Failure Analysis; D004926:Escherichia coli / Q000235:genetics* / Q000302:isolation & purification*; D004950:Esterases / Q000737:chemistry*; D012329:RNA, Bacterial / Q000032:analysis* / Q000235:genetics; D012336:RNA, Ribosomal, 16S / Q000032:analysis* / Q000235:genetics; D014159:Transducers
D050091:Dendrimers; D012329:RNA, Bacterial; D012336:RNA, Ribosomal, 16S; D004950:Esterases