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Omp85 genosensor for detection of human brain bacterial meningitis

Sandip Kumar Dash1, Minakshi Sharma, Shashi Khare

  • 1CSIR-Institute of Genomics and Integrative Biology, Mall Road, Delhi, 110007, India.

Biotechnology Letters
|March 9, 2013
PubMed

Insights

This study presents a novel Omp85 genosensor for rapid Neisseria meningitidis detection. The biosensor accurately identifies bacterial DNA in cerebrospinal fluid, offering a promising tool for meningitis diagnosis.

Area of Science:

  • Biosensor technology
  • Molecular diagnostics
  • Microbiology

Background:

  • Neisseria meningitidis is a major cause of bacterial meningitis.
  • Rapid and accurate diagnostic methods are crucial for effective meningitis treatment.
  • Current diagnostic techniques may require significant time and resources.

Purpose of the Study:

  • To develop and characterize a novel electrochemical genosensor for the specific detection of Neisseria meningitidis.
  • To evaluate the sensitivity, specificity, and stability of the Omp85 genosensor.
  • To assess the potential of the genosensor for diagnosing meningitis directly from cerebrospinal fluid.

Main Methods:

  • Immobilization of a 5'-thiolated DNA probe targeting the Omp85 virulence gene onto a screen-printed gold electrode.
  • Hybridization with Neisseria meningitidis single-stranded genomic DNA (ssG-DNA) from patient cerebrospinal fluid (CSF).
  • Electrochemical detection using cyclic voltammetry, differential pulse voltammetry (DPV), and electrochemical impedance spectroscopy.

Main Results:

  • The Omp85 genosensor demonstrated high sensitivity, detecting as little as 6 ng of ssG-DNA in 6 μl of CSF.
  • The assay provided results within 30 minutes, including a 1-minute response time.
  • The genosensor exhibited good stability, maintaining performance for 12 months at 4°C.
  • DPV analysis showed a sensitivity of 2.6 (μA/cm(2))/ng with a high regression coefficient (R(2) = 0.954).

Conclusions:

  • The developed Omp85 genosensor offers a rapid, sensitive, and stable platform for Neisseria meningitidis detection.
  • This biosensor holds significant potential for the early and accurate diagnosis of meningitis directly from clinical samples.
  • The electrochemical approach provides a cost-effective and accessible diagnostic tool for resource-limited settings.

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