Title:	Development of electrochemical biosensor for detection of staphylococcal enterotoxin A.
Authors:	PIMENTA, M. G. R. FURTADO, R. F. BORGES, M. de F. ALVES, C. R.
Affiliation:	Maria Gardeny Ribeiro Pimenta, UECE; ROSELAYNE FERRO FURTADO, CNPAT; MARIA DE FATIMA BORGES, CNPAT; Carlúcio Roberto Alves, UECE.
Date Issued:	2010
Citation:	In: CONGRESSO BRASILEIRO DE BIOTECNOLOGIA, 3., 2010, Fortaleza. Programa e Resumos. Fortaleza: Sociedade Brasileira de Biotecnologia, 2010. p. 131.
Description:	Staphylococcal food poisoning is one of the rnost prevalent causes of gastroe~teritis worldwide, which ís caused by the ingestion of food with p~eformed toxin. Staphylococcal enterotoxrns (SEs)_are low molecular weight proteins produced by some strams of Staphylococcus aureus. lhese enterotoxrns are heat sta~le, résistant to stornach proteases and stable over a wide range of pH. ~owadays, the methods for detect,on of these enterotoxins are rnade by expensive immunological or biochemical kits. ln the last decades, special àttention has been given on the potential of using biosensors for specific analysis by selective binding the iarget cornpounds. ln this context, the detection based on biosensor may be a desirable tool dueto high sensitivity, specificity, reproducibility and low cost. ln this study, were prepared screen-printed electrodes modified with anti-SEA and subsequent blocking by 3% BSA and 500 mM glydne. ln the process of (lification of the electrode was used the self-assernbly rnonolayer technique (SAM). For this, 10 µl 5 mg rnL· 1 protein A was dropped on the electrode, Next step, added 10 µl of 0.5 µg µL- 1 antibody a'nti-Staphylococcal enterotoxin A (lgG), BSA 3% or glycine (500 mM), 10 µl of 0.1 µg µl'' entero.toxin taphylococcal A (SEA), 10 µL of antibody anti-staphylococcal enterotoxin conjugated to peroxídase. lhe solutions were prepared with phosphate. buffer (pH 7.0). Each immobílization procedure was reafized Jor one hour with subsequent washes in distilled water. Cyclic voltammetry measures were performed in Potentiostat/GalvanostatAutolab PGSTAllO0. lhe glass cell of 10 ml was used anda conventional system 'th three electrodes: a platinum auxiliary electrode, a reference electrode of Ag/AgCI saturated with KG 3 M and a screen-printed electrode as working electrode. The biosensor showed good linearity in phosphate buffer. lhe cyclic voltamrnetry demonstrated good stability of biosensor after fifty scans. Prelirninary studies indicated that 3% BSA was better blocking agent than 500 rnM glycine. lhe choose of the blocking agent important for the developrnent of biosensor_more sensitive and specific. Until now, the results are very omising for the development of biosensor for detection of staphylococcal enterotoxin in food.
Keywords:	Biossenssor
Type of Material:	Resumo em anais e proceedings
Access:	openAccess
Appears in Collections:	Resumo em anais de congresso (CNPAT)