Journal article
A Cantilever Biosensor-Based Assay for Toxin-Producing Cyanobacteria Microcystis aeruginosa using 16S rRNA
Environmental science & technology, v 47(21), pp 12333-12341
05 Nov 2013
PMID: 24070168
Featured in Collection : UN Sustainable Development Goals @ Drexel
Abstract
Monitoring of cyanotoxins in source waters is currently done through toxin-targeting assays which suffer from low sensitivity due to poor antibody avidity. We present a biosensor-based method as an alternative for detecting toxin-producing cyanobacteria M. aeruginosa via species-selective region of 16S rRNA at concentrations as low as 50 cells/mL, and over a five-log dynamic range. The cantilever biosensor was immobilized with a 27-base DNA strand that is complementary to the target variable region of 16S rRNA of M. aeruginosa. The cantilever sensor detects mass-changes through shifts in its resonant frequency. Increase in the biosensor's effective mass, caused by hybridization of target strand with the biosensor-immobilized complementary strand, showed consistent and proportional frequency shift to M. aeruginosa concentrations. The sensor hybridization response was verified in situ by two techniques: (a) presence of duplex DNA structure postdetection via fluorescence measurements, and (b) secondary hybridization of nanogold-labeled DNA strands to the captured 16S rRNA strands. The biosensor-based assay, conducted in a flow format (similar to 0.5mL/min), is relatively short, and requires a postextraction analysis time of less than two hours. The two-step detection protocol (primary and secondary hybridization) is less prone to false negatives, and the technique as a whole can potentially provide an early warning for toxin presence in source waters.
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Details
- Title
- A Cantilever Biosensor-Based Assay for Toxin-Producing Cyanobacteria Microcystis aeruginosa using 16S rRNA
- Creators
- Blake N. Johnson - Drexel UniversityRaj Mutharasan - Drexel Univ, Dept Chem & Biol Engn, Philadelphia, PA 19104 USA
- Publication Details
- Environmental science & technology, v 47(21), pp 12333-12341
- Publisher
- American Chemical Society; Washington, DC
- Number of pages
- 9
- Grant note
- R833829 / EPA STAR Grant; United States Environmental Protection Agency
- Resource Type
- Journal article
- Language
- English
- Academic Unit
- Chemical and Biological Engineering
- Web of Science ID
- WOS:000326711300057
- Scopus ID
- 2-s2.0-84887875863
- Other Identifier
- 991019170854404721
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- Web of Science research areas
- Engineering, Environmental
- Environmental Sciences