Applied Biochemistry and Biotechnology

, Volume 171, Issue 1, pp 198–208

rmpM Genosensor for Detection of Human Brain Bacterial Meningitis in Cerebrospinal Fluid

  • Sandip Kumar Dash
  • Minakshi Sharma
  • Shashi Khare
  • Ashok Kumar
Article

DOI: 10.1007/s12010-013-0339-3

Cite this article as:
Dash, S.K., Sharma, M., Khare, S. et al. Appl Biochem Biotechnol (2013) 171: 198. doi:10.1007/s12010-013-0339-3
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Abstract

Human brain bacterial meningitis is a life-threatening disease caused mainly by Neisseria meningitidis, lead to damage of the outer membrane covering (meninges) of brain or even death. The usual methods of diagnosis are either time-consuming or have some limitations. The specific rmpM (reduction-modifiable protein M) virulent gene based genosensor is more sensitive, specific, and can detect N. meningitidis directly from the patient cerebrospinal fluid in 30 min including 1-min response time. 5′-Thiol-labeled single-stranded DNA (ssDNA) probe was immobilized onto screen-printed gold electrode (SPGE) and hybridized with denatured (95 °C) single-stranded genomic DNA (ssG-DNA) for 10 min at 25 °C. The electrochemical response was measured by cyclic voltammetry, differential pulse voltammetry (DPV) and electrochemical impedance using redox indicators. The sensitivity of the genosensor was 9.5087 (μA/cm2)/ng with DPV and limit of detection was 3 ng/6 μL ssG-DNA. The immobilization of the ssDNA probe and hybridization with ssG-DNA from N. meningitidis was characterized by atomic force microscopy and Fourier transform infrared spectroscopy. The rmpM genosensor was stable for 6 months at 4 °C with 10 % loss in initial DPV current. The advantage of rmpM genosensor is to detect bacterial meningitis simultaneously in multiple patients using SPGE array during an outbreak of the disease.

Keywords

Bacterial meningitis Genosensor Neisseria meningitidis rmpM genosensor 

Copyright information

© Springer Science+Business Media New York 2013

Authors and Affiliations

  • Sandip Kumar Dash
    • 1
  • Minakshi Sharma
    • 2
  • Shashi Khare
    • 3
  • Ashok Kumar
    • 1
  1. 1.CSIR-Institute of Genomics and Integrative BiologyDelhiIndia
  2. 2.Department of ZoologyM. D. UniversityRohtakIndia
  3. 3.National Centre for Disease ControlDelhiIndia

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