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Nanostructured platform for the detection of Neisseria gonorrhoeae using electrochemical impedance spectroscopy and differential pulse voltammetry

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Abstract

We report on a nanocomposite based genosensor for the detection of Neisseria gonorrhoeae, a bacterium causing the sexually transmitted disease gonorrhoea. Amino-labeled probe DNA was covalently immobilized on electrochemically prepared polyaniline and iron oxide (PANI-Fe3O4) nanocomposite film on an indium tin oxide (ITO) electrode. Scanning electron microscopy, transmission electron microscopy, electrochemical impedance spectroscopy (EIS) and differential pulse voltammetry (DPV) techniques have been employed to characterize surface of the modified electrode. The genosensor has detection limits of 1 × 10-15 M and 1 × 10-17 M, respectively, using the EIS and DPV techniques. This biosensor can discriminate a complementary sequence from a single-base mismatch and from non-complementary DNA, and has been utilized for detection of DNA extracted from N. gonorrhoeae culture, and from patient samples with N. gonorrhoeae. It is found to exhibit good specificity for N. gonorrhoeae species and shows no response towards non-gonorrhoeae type of Neisseria species (NgNs) and other gram-negative bacterias (GNBs). The affinity constant for hybridization calculated using the Langmuir adsorption isotherm model is found to be 3.39 × 108 M-1.

Electrochemically deposited nanostructured platform for Neisseria gonorrhoeae detection.

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Acknowledgements

The authors thank Director, National Physical Laboratory, New Delhi, India for the facilities. Renu Singh and Zimple Matharu are thankful to Council of Scientific and Industrial Research (CSIR), India for the award of Senior Research Fellowships. The authors thank Mr. K. N. Sood and Mr. Jai Tawale, NPL for SEM measurements. We thank Dr. G. Sumana, Mrs. Rachna verma, Dr. V. K. Sharma, Professor and Head, Dermatology, AIIMS, Dr. J.C. Samantaray, Professor and Head and Dr. Arti Kapil, Professor, Microbiology, AIIMS, Dr. Manju Bala, Senior Microbiologist, Safdarjang hospital, New Delhi for useful discussions. We acknowledge the financial support received from DST [DST/TSG/ME/2008/18 and GAP-070932], in-house project (OLP-070632D) and Department of Biotechnology, Govt. of India (DBT/GAP070832).

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Authors and Affiliations

  1. Department of Science & Technology Centre on Biomolecular Electronics, Biomedical Instrumentation Section, National Physical Laboratory (Council of Scientific & Industrial Research), Dr. K.S. Krishnan Marg, New Delhi, 110012, India

    Renu Singh, Zimple Matharu, Avanish Kumar Srivastava & Bansi Dhar Malhotra

  2. Department of Microbiology, All India Institute of Medical Sciences, New Delhi, 110002, India

    Seema Sood

  3. School of Biotechnology, Guru Gobind Singh Indraprastha University, Kashmere Gate, Delhi, 110006, India

    Renu Singh & Rajinder Kumar Gupta

  4. Department of Biotechnology, Delhi Technological University, Shahbad Daulatpur, Main Bawana Road, Delhi, 110042, India

    Bansi Dhar Malhotra

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  1. Renu Singh
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  2. Zimple Matharu
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  3. Avanish Kumar Srivastava
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  4. Seema Sood
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Correspondence to Bansi Dhar Malhotra.

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Singh, R., Matharu, Z., Srivastava, A.K. et al. Nanostructured platform for the detection of Neisseria gonorrhoeae using electrochemical impedance spectroscopy and differential pulse voltammetry. Microchim Acta 177, 201–210 (2012). https://doi.org/10.1007/s00604-012-0765-x

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