Hosea Hosea A, Philip Joseph Y N, Maeda Daniel G, Mahadhy Ally
Government Chemist Laboratory Authority, P. O. Box 2925, Dodoma, Tanzania.
Department of Chemistry, University of Dar es Salaam, P. O. Box 35179, Da es Salaam, Tanzania.
Appl Biochem Biotechnol. 2025 Feb;197(2):1242-1257. doi: 10.1007/s12010-024-05088-x. Epub 2024 Nov 20.
Canine distemper, a viral disease with a global impact on various animals including dogs, foxes, wolves, lions, and leopards, requires early diagnosis for effective treatment and outbreak control. Common laboratory methods, such as enzyme-linked immunosorbent assay, polymerase chain reaction, and viral isolation, face challenges such as extended turnaround times, high costs, and the expertise required. This study has developed a field-based biosensor for detecting the canine distemper virus (CDV), utilising a screen-printed carbon electrode (SPCE) and a computer-assisted portable potentiostat. A 30-mer oligonucleotide capture probe, designed using Primer3 Plus software version 3.3.0, detected hybridisation with the CDV complementary strand through electrochemical analysis via differential pulsed voltammetry. The developed biosensor exhibited good linearity in quantifying the target analyte concentration (0.1 to 12.8 µM), with a detection limit of 0.05 µM, indicating high sensitivity. Specificity tests using complementary and non-complementary sequences confirmed the biosensor's accuracy. The electrode can be reused up to eight times with a residual capacity of 93.72 ± 5.45% after regeneration using a 50 mM NaOH solution. The developed biosensor was also used to detect CDV in biological samples after RNA extraction and amplification. Results from the biosensor aligned with those from reverse transcriptase polymerase chain reaction (RT-PCR) findings, showing 100% agreement. These findings support the potential development of a field-deployable portable device for effectively diagnosing canine distemper in biological samples.
犬瘟热是一种对包括狗、狐狸、狼、狮子和豹子在内的各种动物具有全球影响的病毒性疾病，需要早期诊断以进行有效治疗和疫情控制。常见的实验室方法，如酶联免疫吸附测定、聚合酶链反应和病毒分离，面临着周转时间长、成本高以及所需专业知识等挑战。本研究开发了一种基于现场的生物传感器，用于检测犬瘟热病毒（CDV），该传感器利用了丝网印刷碳电极（SPCE）和计算机辅助便携式恒电位仪。使用Primer3 Plus软件版本3.3.0设计的30聚体寡核苷酸捕获探针，通过差分脉冲伏安法进行电化学分析，检测与CDV互补链的杂交。所开发的生物传感器在定量目标分析物浓度（0.1至12.8 μM）方面表现出良好的线性，检测限为0.05 μM，表明具有高灵敏度。使用互补和非互补序列的特异性测试证实了生物传感器的准确性。该电极可重复使用多达八次，使用50 mM NaOH溶液再生后，剩余容量为93.72±5.45%。所开发的生物传感器还用于在RNA提取和扩增后检测生物样品中的CDV。生物传感器的结果与逆转录聚合酶链反应（RT-PCR）的结果一致，显示出100%的一致性。这些发现支持了开发一种可现场部署的便携式设备以有效诊断生物样品中犬瘟热的潜力。
