Attomolar electrochemical detection of the BCR/ABL fusion gene based on an amplifying self-signal metal nanoparticle-conducting polymer hybrid composite.
In the last ten years, conjugated polymers started to be used in the immobilization of nucleic acids via non-covalent interactions. In the present study, we describe the construction and use of an electrochemical DNA biosensor based on a nanostructured polyaniline-gold composite, specifically developed for the detection of the BCR/ABL chimeric oncogene. This chromosome translocation is used as a biomarker to confirm the clinical diagnosis of both chronic myelogenous leukemia (CML) and acute lymphocytic leukemia (ALL). The working principle of the biosensor rests on measuring the conductivity resulting from the non-covalent interactions between the hybrid nanocomposite and the DNA probe. The nanostructured platform exhibits a large surface area that enhances the conductivity. Positive cases, which result from the hybridization between DNA probe and targeted gene, induce changes in the amperometric current and in the charge transfer resistance (RCT) responses. Atomic force microscopy (AFM) images showed changes in the genosensor surface after exposure to cDNA sample of patient with leukemia, evidencing the hybridization process. This new hybrid sensing-platform displayed high specificity and selectivity, and its detection limit is estimated to be as low as 69.4 aM. The biosensor showed excellent analytical performance for the detection of the BCR/ABL oncogene in clinical samples of patients with leukemia. Hence, this electrochemical sensor appears as a simple and attractive tool for the molecular diagnosis of the BCR/ABL oncogene even in early-stage cases of leukemia and for the monitoring of minimum levels of residual disease.
Programa de Pós-Graduação em Bioquímica e Fisiologia, Universidade Federal de Pernambuco, 50670-901 Recife, PE, Brazil.
Karen Y P S
KYPS
Avelino
Programa de Pós-Graduação em Inovação Terapêutica, Universidade Federal de Pernambuco, 50670-901 Recife, PE, Brazil.
Isaac A M
IAM
Frias
Centro de Pesquisas Aggeu Magalhães, Fundação Osvaldo Cruz (Fiocruz), 50670-420 Recife, PE, Brazil; Laboratório de Biologia Molecular, Departamento de Oncologia Pediátrica, Instituto de Medicina Integral Professor Fernando Figueira (IMIP), 50070-550 Recife, PE, Brazil.
Norma
N
Lucena-Silva
Centro de Pesquisas Aggeu Magalhães, Fundação Osvaldo Cruz (Fiocruz), 50670-420 Recife, PE, Brazil; Laboratório de Biologia Molecular, Departamento de Oncologia Pediátrica, Instituto de Medicina Integral Professor Fernando Figueira (IMIP), 50070-550 Recife, PE, Brazil.
Renan G
RG
Gomes
Departamento de Física, Universidade Federal de Pernambuco, 50670-901 Recife, PE, Brazil.
Celso P
CP
de Melo
Programa de Pós-Graduação em Bioquímica e Fisiologia, Universidade Federal de Pernambuco, 50670-901 Recife, PE, Brazil; Programa de Pós-Graduação em Inovação Terapêutica, Universidade Federal de Pernambuco, 50670-901 Recife, PE, Brazil.
Maria D L
MDL
Oliveira
Programa de Pós-Graduação em Bioquímica e Fisiologia, Universidade Federal de Pernambuco, 50670-901 Recife, PE, Brazil; Programa de Pós-Graduação em Inovação Terapêutica, Universidade Federal de Pernambuco, 50670-901 Recife, PE, Brazil. Electronic address: csrandrade@pq.cnpq.br.
César A S
CAS
Andrade
D000814:Aniline Compounds / Q000737:chemistry; D015374:Biosensing Techniques / Q000379:methods; D015342:DNA Probes / Q000737:chemistry / Q000235:genetics; D004273:DNA, Neoplasm / Q000737:chemistry / Q000235:genetics; D004553:Electric Conductivity; D055664:Electrochemical Techniques / Q000379:methods; D016044:Fusion Proteins, bcr-abl / Q000235:genetics*; D006046:Gold / Q000737:chemistry; D006801:Humans; D007938:Leukemia / Q000175:diagnosis / Q000235:genetics; D053768:Metal Nanoparticles / Q000737:chemistry*; D018625:Microscopy, Atomic Force; D053761:Nanocomposites / Q000737:chemistry*; D016133:Polymerase Chain Reaction; D011108:Polymers / Q000737:chemistry*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D000814:Aniline Compounds; D015342:DNA Probes; D004273:DNA, Neoplasm; D011108:Polymers; C416807:polyaniline; D006046:Gold; D016044:Fusion Proteins, bcr-abl
BCR/ABL fusion gene; Biosensor; Electrochemistry; Hybrid nanoparticles; Leukemia