Electrochemical DNA biosensor for bovine papillomavirus detection using polymeric film on screen-printed electrode.
A new electrochemical DNA biosensor for bovine papillomavirus (BPV) detection that was based on screen-printed electrodes was comprehensively studied by electrochemical methods of cyclic voltammetry (CV) and differential pulse voltammetry (DPV). A BPV probe was immobilised on a working electrode (gold) modified with a polymeric film of poly-L-lysine (PLL) and chitosan. The experimental design was carried out to evaluate the influence of polymers, probe concentration (BPV probe) and immobilisation time on the electrochemical reduction of methylene blue (MB). The polymer poly-L-lysine (PLL), a probe concentration of 1 μM and an immobilisation time of 60 min showed the best result for the BPV probe immobilisation. With the hybridisation of a complementary target sequence (BPV target), the electrochemical signal decreased compared to a BPV probe immobilised on the modified PLL-gold electrode. Viral DNA that was extracted from cattle with papillomatosis also showed a decrease in the MB electrochemical reduction, which suggested that the decreased electrochemical signal corresponded to a bovine papillomavirus infection. The hybridisation specificity experiments further indicated that the biosensor could discriminate the complementary sequence from the non-complementary sequence. Thus, the results showed that the development of analytical devices, such as a biosensor, could assist in the rapid and efficient detection of bovine papillomavirus DNA and help in the prevention and treatment of papillomatosis in cattle.
Laboratório de Imunopatologia Keizo Asami (LIKA), Universidade Federal de Pernambuco-UFPE, Av. Prof. Moraes Rego, s/n, Campus Universitário, 50670-901 Recife, PE, Brazil. galvesn23@gmail.com
Gustavo A
GA
Nascimento
Elaine V M
EV
Souza
Danielly S
DS
Campos-Ferreira
Mariana S
MS
Arruda
Carlos H M
CH
Castelletti
Marcela S O
MS
Wanderley
Marek H F
MH
Ekert
Danyelly
D
Bruneska
José L
JL
Lima-Filho
D000818:Animals; D015374:Biosensing Techniques / Q000379:methods*; D052688:Bovine papillomavirus 1 / Q000302:isolation & purification*; D002417:Cattle / Q000821:virology*; D048271:Chitosan / Q000737:chemistry; D004279:DNA, Viral / Q000032:analysis*; D055664:Electrochemical Techniques / Q000379:methods*; D004566:Electrodes; D006046:Gold / Q000737:chemistry; D008751:Methylene Blue / Q000737:chemistry; D009693:Nucleic Acid Hybridization / Q000379:methods; D010084:Oxidation-Reduction; D030361:Papillomavirus Infections / Q000175:diagnosis; D011107:Polylysine / Q000737:chemistry; D012680:Sensitivity and Specificity
D004279:DNA, Viral; D011107:Polylysine; D006046:Gold; D048271:Chitosan; D008751:Methylene Blue