Ultrasensitive immunoassay for detection of Citrus tristeza virus in citrus sample using disposable microfluidic electrochemical device

Autor(es)

Freitas, Tayane A. ^[1] ; Proenca, Camila A. ^[1] ; Baldo, Thaisa A. ^[1] ; Materon, Elsa M. ^[1] ; Wong, Ademar ^[1] ; Magnani, Rodrigo F. ^{[1, 2]} ; Faria, Ronaldo C. ^[1]

Afiliação do(s) autor(es)

^[1] Univ Fed Sao Carlos, Dept Chem, BR-13565905 Sao Carlos, SP - Brazil

^[2] Fundecitrus, Dept Pesquisa & Desenvolvimento, BR-14807040 Araraquara, SP - Brazil

• Tipo de documento: Artigo Científico
• Fonte: Talanta; v. 205, DEC 1 2019.
• Citações Web of Science: 2

Resumo

Tristeza is a disease that affects citrus crops in general, caused by the Citrus tristeza virus (CTV). It is considered an economically important virus diseases in citrus, which is present in the main citrus producing regions all around the world. Early detection of CTV is crucial to avoid any epidemics and substantial economic losses for the citrus growers. Consequently, the development of rapid, accurate, and sensitive methods capable of detecting the virus in the early stages of the disease is highly desired. Based on that, a low-cost and rapid magneto-immunoassay methodology to detect the capsid protein from CTV (CP-CTV) was proposed. For this, magnetic beads were decorated with antibodies anti-CP-CTV and horseradish peroxidase enzyme (HRP) and applied for the capture and separation of CP-CTV from the sample solutions. The magnetically captured biomarker was detected using a simple disposable microfluidic electrochemical device (D mu FED) constructed by rapid prototyping technique and composed by an array of immunosensors. In D mu FED, the electrodes were modified with monoclonal antibody anti-CP-CTV and the detection was carried out using amperometry, based on the hydroquinone/H2O2 catalytic redox reaction due to the presence of HRP label in an immune-sandwich structure. The proposed immunoassay presented excellent linearity with a wide linear range of concentration of 1.95-10.0 x 10(3) fg mL(-1) and ultralow detection limit of 0.3 fg mL(1). The disposable device was successfully applied for the detection of Citrus tristeza virus in healthy and infected plant samples, where it showed good agreements with the comparative method of enzyme-linked immunosorbent assay (ELISA). The developed immunoassay methodology showed a sensitive and selective way in the detection of CTV. Hence, it can be considered as a promising analytical alternative for rapid and low-cost diagnosis of Tristeza disease in citrus. (AU)

• Processo FAPESP: 17/24053-7 - Desenvolvimentos de métodos eletroquímicos simples para a detecção de biomarcadores visando o diagnóstico de doenças
• Beneficiário: Ronaldo Censi Faria
• Modalidade de apoio: Auxílio à Pesquisa - Regular
• Processo FAPESP: 16/00991-5 - Estudo da Interação de Enzimas de Detoxificação com Fármacos Quimioterápicos e sua relação com a Quimiorresistência
• Beneficiário: Elsa María Materón Vásques
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado
• Processo FAPESP: 15/19890-1 - Desenvolvimento de métodos para diagnóstico de doenças utilizando partículas magnéticas e arranjo de imunossensores em sistema microfluídico descartável
• Beneficiário: Ronaldo Censi Faria
• Modalidade de apoio: Auxílio à Pesquisa - Regular