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Carbon composite-based DNA sensor for detection of bacterial meningitis caused by Neisseria meningitidis

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Abstract

Carbon/1-octadecanethiol-carboxylated multiwalled carbon nanotubes (cMWCNT) composite was used to construct a DNA sensor for detection of human bacterial meningitis caused by Neisseria meningitidis. The carbon composite electrode was used to covalently immobilize 5′-amine-labeled 19-mer single-stranded DNA (ssDNA) probe, which was hybridized with 1.35 × 102–3.44 × 104 pM (0.5–128 ng/5 μl) of single-stranded genomic DNA (ssG-DNA) of N. meningitidis for 10 min at room temperature (RT). The surface topography of the DNA sensor was characterized by using scanning electron microscopy (SEM) and Fourier transform infrared spectroscopy (FTIR) while electrochemically characterized by electrochemical impedance. The immobilization of ssDNA probe and hybridization with ssG-DNA were detected electrochemically by cyclic voltammetry (CV) and differential pulse voltammetry (DPV) at RT in 30 min with a response time of 1 min. The DNA sensor showed high pathogenic specificity and can distinguish among complement, noncomplement, one base mismatch, and triple base mismatch oligomer targets. The limit of detection (LOD) and sensitivity of the sensor were approximately 68 pM and 38.095 (μA/cm2)/nM of ssG-DNA, respectively, using DPV. The improved sensitivity and LOD of the sensor can be attributed to the higher efficiency of probe immobilization due to high surface area-to-volume ratio and good electrical activity of cMWCNT.

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Acknowledgment

S.K. Dash thanks University Grant Commission for providing senior research fellowship.

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Authors and Affiliations

  1. Council of Scientific and Industrial Research (CSIR)-Institute of Genomics and Integrative Biology, Mall Road, New Delhi, Delhi, 110007, India

    Sandip K. Dash, Akhilesh Kumar & Ashok Kumar

  2. Department of Zoology, Maharshi Dayanand University, Rohtak, Haryana, 124001, India

    Minakshi Sharma

  3. National Centre for Disease Control, Sham Nath Marg, New Delhi, Delhi, 110054, India

    Shashi Khare

Authors
  1. Sandip K. Dash
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  2. Minakshi Sharma
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  3. Akhilesh Kumar
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  4. Shashi Khare
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  5. Ashok Kumar
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Corresponding author

Correspondence to Ashok Kumar.

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Fig. S1

UV spectra of MWCNT at 190-320 nm A after sonication in H2SO4:HNO3 (3:1, v/v) for 0, 4, 6, 8, 10, and 12 h (a-f), B after sonication in autoclaved ddH2O for 0, 4, 6, 8, and 12 h (a-e) (PDF 81 kb)

Fig. S2

A Cyclic voltammogram of carbon (a) and carbon/ODT-cMWCNT composite (b), B Differential pulse voltammogram of carbon (a) and carbon/ODT-cMWCNT composite (b) (PDF 158 kb)

Fig. S3

Cyclic voltammogram of carbon/ODT-MWCNT/ssDNA (a), after hybridization with 4.3x103 pM of ssG-DNA of N. meningitidis for 2, 5, 10, 15, and 20 min (b-f) (PDF 166 kb)

Fig. S4

A Plot between CV relative I pc with respect to carbon/ODT-MWCNT/ssDNA and different concentrations of hybridizing ssG-DNA (0.0-3.44 × 104 pM) from Fig. 4B inset, inset shows 0.0-5.40 × 102 pM region of the plot for calculation of the slope, sensitivity, and LOD of the sensor using CV. B Plot between DPV relative I p with respect to carbon/ODT-MWCNT/ssDNA and different concentrations of hybridizing ssG-DNA (0.0-3.44 × 104 pM) from Fig. 5A inset, inset shows 0.0-5.40 × 102 pM region of the plot for calculation of slope, sensitivity, and LOD of the sensor using DPV (PDF 218 kb)

Fig. S5

Plot between % I p (DPV) of carbon/ODT-MWCNT/ssDNA and time measured over a period of 180 days at intervals of 15 days, the values shown are averages of three readings taken at same conditions (PDF 112 kb)

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Dash, S.K., Sharma, M., Kumar, A. et al. Carbon composite-based DNA sensor for detection of bacterial meningitis caused by Neisseria meningitidis . J Solid State Electrochem 18, 2647–2659 (2014). https://doi.org/10.1007/s10008-014-2525-9

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  • DOI: https://doi.org/10.1007/s10008-014-2525-9

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