International journal of biological macromolecules · 2017
Chitosan stabilized gold nanoparticle mediated self-assembled gliP nanobiosensor for diagnosis of Invasive Aspergillosis.
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Bhatnagar Ira, Mahato Kuldeep, Ealla Kranthi Kiran Reddy, Asthana Amit, Chandra Pranjal
Аннотация
Diagnosis of Invasive Aspergillosis (IA) casused by Aspergillus fumigatus in miniaturized setting is challenging with great importance in human health. In this direction, we have designed a sensitive electrochemical nanobiosensor for diagnosis of IA through detecting the virulent glip target gene (glip-T) in a miniaturized experimetal setting. The sensor probe was fabricated using 1,6-Hexanedithiol and chitosan stabilized gold nanoparticle mediated self-assembly of glip probes (glip-P) on gold electrode. It was characterized by UV-visible spectroscopy, cyclic voltametry and electrochemical impedance spectroscopy. The ability of sensor to detect glip-T was analysed based on the hybridyzation reaction and the signal obtained using toluidine blue as indicator molecule. Analytical parameters were optimized in terms of glip-P concentration, temperature, reaction time, and concentration of toluidine blue. The biosensor showed the dynamic range between 1 × 10-14- 1 × 10-2 M with the detection limit of 0.32 ± 0.01 × 10-14(RSD < 5.2%). The regeneration of biosensor was evaluated and the interference due to non-target oligonucleotide sequences was evaluated individualy as well as in mixed sample to validate the high selectivity of the designed sensor. The stability of the designed sensor was examined and practical applicability of biosensor was tested by detecting glip-T in real sample environment.