Electrochemical DNA biosensor for the detection of human papillomavirus E6 gene inserted in recombinant plasmid

被引:28
作者
Campos-Ferreira, Danielly S. [1 ]
Souza, Elaine V. M. [1 ]
Nascimento, Gustavo A. [1 ]
Zanforlin, Deborah M. L. [1 ]
Arruda, Mariana S. [1 ]
Beltrao, Monique F. S. [1 ]
Melo, Aila L. [1 ]
Bruneska, Danyelly [1 ,2 ]
Lima-Filho, Jose L. [1 ,2 ]
机构
[1] Univ Fed Pernambuco UFPE, LIKA, Ave Prof Moraes S-N, BR-50670901 Recife, PE, Brazil
[2] Univ Fed Pernambuco UFPE, Dept Bioquim, Ave Prof Moraes Rego S-N, BR-50670901 Recife, PE, Brazil
关键词
Electrochemical DNA biosensor; Direct DNA detection; Inosine-probe; Human papillomavirus 16; Electrophoresis standard method; PENCIL GRAPHITE ELECTRODE; HUMAN INTERLEUKINE-2 GENE; HYBRIDIZATION BIOSENSOR; HPV INFECTION; REAL SAMPLES; AMES TEST; OLIGONUCLEOTIDE; ACID; VACCINES; SENSORS;
D O I
10.1016/j.arabjc.2014.05.023
中图分类号
O6 [化学];
学科分类号
0703 ;
摘要
In the current study, we describe a novel, simple, inexpensive, sensitive, specific, stable and label-free electrochemical DNA biosensor used to identify a target gene cloned into a plasmid. The biosensor was designed with a 23-mer oligonucleotide of guanine-free, which was immobilized on the pencil graphite electrode (PGE) for E6 gene detection from human papillomavirus 16 type (HPV16). The E6 gene was used due to its clinical importance. The optimal probe concentration was obtained in 500 nM. The hybridization detection showed a good linearity in the range of 40-5,000 pg/mu L with a detection limit of 16 pg/mu L. The electrochemical method showed higher sensitivity and specificity when compared with the agarose gel electrophoresis assay. This technology could be postulated as a new and attractive alternative for cloning analysis in plasmids. (C) 2014 Production and hosting by Elsevier B.V. on behalf of King Saud University.
引用
收藏
页码:443 / 450
页数:8
相关论文
共 65 条
[1]  
[Anonymous], 2000, DESIGN ANAL EXPT
[2]  
Armbruster David A, 2008, Clin Biochem Rev, V29 Suppl 1, pS49
[3]  
Azam F, 2010, J PAK MED ASSOC, V60, P676
[4]   Aligned carbon nanotube thin films for DNA electrochemical sensing [J].
Berti, F. ;
Lozzi, L. ;
Palchetti, I. ;
Santucci, S. ;
Marrazza, G. .
ELECTROCHIMICA ACTA, 2009, 54 (22) :5035-5041
[5]   The role of inflammation in HPV carcinogenesis [J].
Boccardo, Enrique ;
Lepique, Ana P. ;
Villa, Luisa L. .
CARCINOGENESIS, 2010, 31 (11) :1905-1912
[6]   Human papillomavirus genome variants [J].
Burk, Robert D. ;
Harari, Ariana ;
Chen, Zigui .
VIROLOGY, 2013, 445 (1-2) :232-243
[7]   Global Analysis of Extracytoplasmic Stress Signaling in Escherichia coli [J].
Bury-Mone, Stephanie ;
Nomane, Yanoura ;
Reymond, Nancie ;
Barbet, Romain ;
Jacquet, Eric ;
Imbeaud, Sandrine ;
Jacq, Annick ;
Bouloc, Philippe .
PLOS GENETICS, 2009, 5 (09)
[8]   Electrochemical DNA biosensor for human papillomavirus 16 detection in real samples [J].
Campos-Ferreira, Danielly S. ;
Nascimento, Gustavo A. ;
Souza, Elaine V. M. ;
Souto-Maior, Maria A. ;
Arruda, Mariana S. ;
Zanforlin, Deborah M. L. ;
Ekert, Marek H. F. ;
Bruneska, Danyelly ;
Lima-Filho, Jose L. .
ANALYTICA CHIMICA ACTA, 2013, 804 :258-263
[9]   DETECTION OF ENZYMATICALLY AMPLIFIED HUMAN IMMUNODEFICIENCY VIRUS-DNA BY OLIGONUCLEOTIDE SOLUTION HYBRIDIZATION AND BY INCORPORATION OF RADIOLABELED DEOXYNUCLEOTIDES [J].
CARMAN, WF ;
WILLIAMSON, C .
JOURNAL OF CLINICAL MICROBIOLOGY, 1989, 27 (11) :2570-2573
[10]   HPV infection and cervical disease: A review [J].
Carter, Jonathan R. ;
Ding, Zongqun ;
Rose, Barbara R. .
AUSTRALIAN & NEW ZEALAND JOURNAL OF OBSTETRICS & GYNAECOLOGY, 2011, 51 (02) :103-108