Research Article
Electrochemical DNA Biosensor for Sequences Related to the Human Papillomavirus Type 16 using Methylene Blue
Elaine VM Souza1,3*, Gustavo A Nascimento1, Nataly Amorim de Santana1, Danielly S. Campos-Ferreira1, Juliana de Araújo Bibiano1, Mariana S Arruda1, Danyelly Bruneska1,2 and José L Lima-Filho1,2
1Laboratory of Immunopathology Keizo Asami (LIKA), Federal University of Pernambuco (UFPE), AV. Moraes s/n, 50670-901 Recife, PE, Brazil
2Department of Biochemistry, Federal University of Pernambuco (UFPE), AV. 12 Professor Moraes Rego s/n, 50670-901 Recife, PE, Brazil
3Nursing Department, Universidade Federal de Alagoas-UFAL, Arapiraca 15 Campus, AV. Manoel Severino Barbosa, s/n, Bom Sucesso-Arapiraca-AL, CEP:57309-005, Brazil
- Corresponding Author:
- Souza EVM
1 Laboratory of Immunopathology Keizo Asami (LIKA)
Federal University of Pernambuco (UFPE)
AV. Moraes s/n, 50670-901 Recife, PE, Brazil
Tel: +55 82 9931 0827
Fax: 82-3214 +55 1172
E-mail: elainevms@yahoo.com.br
Received Date: May 10, 2014; Accepted Date: July 28, 2014; Published Date: August 05, 2014
Citation: Souza EVM, Nascimento GA, Santana NA, Campos-Ferreira DS, Bibiano JA, et al. (2014) Electrochemical DNA Biosensor for Sequences Related to the Human Papillomavirus Type 16 using Methylene Blue. Biosens J 3:107. doi: 10.4172/2090-4967.1000107
Copyright: © 2014 Souza EVM, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Abstract
Several studies show that infections with high-risk human papillomaviruses (HPV), mainly HPV type 16, can lead to the development of tumors as the cervical cancer. A rapid and precise diagnosis of the precancerous lesions by HPV is extremely important for a successful treatment. The present paper describes an electrochemical DNA biosensor for specific sequence detection of the E1 HPV 16, using the methylene blue (MB) as indicator hybridization. In order to develop the sensor, a 21-mer ssDNA probe related to E1 HPV gene was immobilized on the work electrode (WE). The obtained optimum conditions for immobilization of HPVE1.16P on the activated WE was with an immobilization time of 5 min and probe concentration of 8 μM. The hybridization between the probe and its complementary sequence was studied by Differential Pulse Voltammetry (DPV). Some hybridization experiments with non-complementary oligonucleotides were carried out to study the selectively of the biosensor. The results showed that this DNA biosensor could be used for detection E1 HPV gene due to selectivity of the electrochemical measurement system. A detection limit of probe immobilized on electrode to its complementary sequence was 1.49 nM.