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7X multiplexed, optofluidic detection of nucleic acids for antibiotic-resistance bacterial screening - PubMed

  • ️Wed Jan 01 2020

7X multiplexed, optofluidic detection of nucleic acids for antibiotic-resistance bacterial screening

G G Meena et al. Opt Express. 2020.

Abstract

Rapid and accurate diagnosis of bacterial infections resistant to multiple antibiotics requires development of new bio-sensors for differentiated detection of multiple targets. This work demonstrates 7x multiplexed detection for antibiotic-resistance bacterial screening on an optofluidic platform. We utilize spectrally multiplexed multi-spot excitation for simultaneous detection of nucleic acid strands corresponding to bacterial targets and resistance genes. This is enabled by multi-mode interference (MMI) waveguides integrated in an optofluidic device. We employ a combinatorial three-color labeling scheme for the nucleic acid assays to scale up their multiplexing capability to seven different nucleic acids, representing three species and four resistance genes.

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Conflict of interest statement

A.R.H and H.S have financial interest in Fluxus Inc. which is developing optofluidic devices.

Figures

Fig. 1.
Fig. 1.

(a) Schematic view of experimental setup and optofluidic chip. (b) Top down optical microscope image of a chip (Scale bar: 200μm) (c) Color coded fluorescent dye image of the patterns generated by the MMI waveguide in the LC waveguide.

Fig. 2.
Fig. 2.

(a) Schematic image of magnetic-bead based assays with targets combinatorieally labeled with dark red, red and green colors. (b) Color codes used to label the seven different antibiotic resistance gene/species targets [DR-Dark Red, R-Red, G-Green].

Fig. 3.
Fig. 3.

Single-target experiments. (a) E.Coli detection: (i) Detected fluorescence signals when beads with E. coli) targets are flowed through the chip and excited by the MMI waveguide; colored (black) symbols on each event represent correctly (incorrectly) identified targets. The dashed line indicates the threshold for particle detection. (ii) Zoomed-in signal from a single bead. (iii) Bar histogram map of S(t)C of a correctly identified E. coli signal. (b) and (c) same analysis for IMP and E. aerogenes targets.

Fig. 4.
Fig. 4.

(a) Histogram of S(t)C values of all the detected single colored (DR) E. coli-carrying beads. Most of the signals dominate only in the dark red channel ((b) and (c) are from two -color (DR-R) IMP targets and three-color (DR-R-G) E. aerogenes targets).

Fig. 5.
Fig. 5.

Fluorescence signals from all seven bacterial targets are flowed simultaneously through the chip, excited by all three lasers and identified; dashed line: threshold for target detection.

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