Electrocatalytic Detection of Pathogenic DNA Sequences and Antibiotic Resistance Markers

Melissa A. Lapierre, Meaghan O'Keefe, Bradford J. Taft, Shana O. Kelley*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

108 Scopus citations

Abstract

The detection of specific DNA sequences using electrochemical readout would permit the rapid and inexpensive detection and identification of bacterial pathogens. A new assay developed for this purpose is described that harnesses a sensitive electrocatalytic process to monitor DNA hybridization. Two sequences belonging to the pathogenic microbe Helicobacterpylori are used to demonstrate the versatility and specificity of the assay: one that codes for an unique H. pylori protein and one that represents a small portion of the 23S rRNA from this organism. Both sequences can be monitored into the nanomolar concentration range. Target sequences introduced to the electrode surface as synthetic oligonucleotides, PCR products, and RNA transcripts are all detected with high specificity. In addition to reporting the presence of pathogen-related sequences, this assay can accurately resolve single-base changes in target sequences. An A2143C substitution within the H. pylori rRNA that confers antibiotic resistance significantly attenuates hybridization to an immobilized probe corresponding to the WT sequence. The single-base mismatch introduced by this mutation slows the kinetics of hybridization and permits discrimination of the two sequences when short hybridization times are employed. The remarkable sensitivity of this label-free assay to small sequence changes may provide the basis of a new method for the detection and genotyping of infectious bacteria using electrochemical methods.

Original languageEnglish (US)
Pages (from-to)6327-6333
Number of pages7
JournalAnalytical Chemistry
Volume75
Issue number22
DOIs
StatePublished - Nov 15 2003
Externally publishedYes

ASJC Scopus subject areas

  • Analytical Chemistry

Fingerprint

Dive into the research topics of 'Electrocatalytic Detection of Pathogenic DNA Sequences and Antibiotic Resistance Markers'. Together they form a unique fingerprint.

Cite this