title: Omp85 genosensor for detection of human brain bacterial meningitis.
abstract: The 5'-thiolated DNA probe based on specific virulence gene, Omp85, was immobilized onto a screen-printed gold electrode followed by hybridization with 6-100 ng/6 μl (5.9 × 10(5)-9.3 × 10(6 )c.f.u.) of Neisseria meningitidis single stranded genomic DNA (ssG-DNA) for 10 min at 25 °C from the cerebrospinal fluid (CSF) of a meningitis patient. The Omp85 genosensor can detect as little as 6 ng ssG-DNA in 6 μl CSF of a human brain meningitis patient in 30 min including a response time of 1 min by cyclic voltammetry, differential pulse voltammetry (DPV) and electrochemical impedance. The sensitivity of the genosensor electrode was 2.6(μA/cm(2))/ng using DPV with regression coefficient (R(2)) 0.954. The genosensor was characterized using Fourier transform infrared spectroscopy and atomic force microscopy. Omp85 genosensor was stable for 12 months at 4 °C with 12 % loss in DPV current.
authors.affiliation: CSIR-Institute of Genomics and Integrative Biology, Mall Road, Delhi, 110007, India.
authors.forename: Sandip Kumar
authors.initials: SK
authors.lastname: Dash
authors.forename: Minakshi
authors.initials: M
authors.lastname: Sharma
authors.forename: Shashi
authors.initials: S
authors.lastname: Khare
authors.forename: Ashok
authors.initials: A
authors.lastname: Kumar
mesh_terms: D001425:Bacterial Outer Membrane Proteins / Q000235:genetics*; D001431:Bacteriological Techniques / Q000379:methods*; D002555:Cerebrospinal Fluid / Q000382:microbiology*; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D006801:Humans; D008585:Meningitis, Meningococcal / Q000175:diagnosis*; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D008969:Molecular Sequence Data; D009345:Neisseria meningitidis / Q000235:genetics / Q000302:isolation & purification*; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D013997:Time Factors
chemical_list: D001425:Bacterial Outer Membrane Proteins; D004269:DNA, Bacterial; C115176:Omp85 protein, Neisseria