Poonam Nandal, Rahul khatri, Vishakha Aggarwal,Ravina,Minakshi Prasad,Hari Mohan and Chandra S. Pundir
The 5’-thiolated DNA probe of Meq oncogenic gene consisting of 24 deoxynucleotide, was reduced by Dithiothreitol (DTT) and immobilized covalently via thiolated bond onto gold electrode followed by hybridization with 1-4 ng/μl of Marek’s Disease Virus (MDV) Single-Stranded (ss) DNA probe from tumor sample of chicken for 10 min at room temp. The Meqgenosensor could detect 1 ngss-DNA in tumor samples in min by Cyclic Voltammetry (CV), Differential Pulse Voltammetry (DPV) and Electrochemical Impedance (EI). The ssDNA modified Au electrode was characterized by Fourier Transform Infrared Spectroscopy (FTIR) and Scanning Electron Microscopy (SEM). An amperometric genosensor was constructed for detection of Meq gene by connecting ssDNA electrode modified Au electrode (working electrode) with Ag/ AgCl electrode (reference electrode) and Pt wire (as auxiliary electrode) through potentiostat. The hybridization between ssDNA probe and target ssDNA was detected by reduction in current generated by interaction of Methylene Blue (MB) with free G’ of single stranded oligonucleotide. The ssDNA Au electrode showed optimum current within 10 min at pH8, 250C. This is the first report on construction of genosensor for the simple, fast and specific detection of MDV in chickens.
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