Jayanath Nileththi Yasendra, Nguyen Loc Thai, Vu Thu Thi, Tran Lam Dai
Department of Food Science & Technology, Faculty of Agriculture, University of Peradeniya Sri Lanka.
Food Engineering and Bioprocess Technology, School of Environment, Resources and Development (SERD), Asian Institute of Technology PO Box 4, Klong Luang 12120, Pathumthani Bangkok Thailand
RSC Adv. 2018 Oct 12;8(61):34954-34959. doi: 10.1039/c8ra07235c. eCollection 2018 Oct 10.
The objective of this study was to develop a simple, inexpensive prototype device for rapid detection of hepatitis B virus (HBV). The device was able to simultaneously amplify, detect and quantify the target HBV DNA. The system was fabricated from a custom-made electrochemical set-up of which the temperature was thermostatically controlled by a water bath. Real-time monitoring of HBV DNA was accomplished by measuring the response of redox indicator in the reaction mixture. Concentration of HBV DNA in the samples was determined from the peak high ratio (PHR) and threshold time relationship. The signal was processed by sigmoidal model fitting to enhance the accuracy of the results. Key parameters including concentrations of redox indicator and reaction temperatures were optimized. Sensitivity and specificity of the method toward HBV DNA were evaluated. The prototype was capable of real-time amplification and detection of HBV DNA with concentration as low as 6.18 fg μl. The test showed high specificity against HBV DNA. The system was also able to detect HBV positive serum directly with simple thermal pretreatment instead of tedious DNA extraction. The electrochemical set-up was compatible with microfluidic platforms and can be readily adapted for efficient and high throughput point-of-care (POC) diagnosis of HBV.
本研究的目的是开发一种用于快速检测乙型肝炎病毒（HBV）的简单、低成本原型设备。该设备能够同时扩增、检测和定量目标HBV DNA。该系统由定制的电化学装置制成，其温度由水浴恒温控制。通过测量反应混合物中氧化还原指示剂的响应来实现对HBV DNA的实时监测。根据峰高比（PHR）和阈值时间关系确定样品中HBV DNA的浓度。通过S形模型拟合对信号进行处理，以提高结果的准确性。对包括氧化还原指示剂浓度和反应温度在内的关键参数进行了优化。评估了该方法对HBV DNA的灵敏度和特异性。该原型能够实时扩增和检测浓度低至6.18 fg/μl的HBV DNA。该测试对HBV DNA显示出高特异性。该系统还能够通过简单的热预处理直接检测HBV阳性血清，而无需繁琐的DNA提取。该电化学装置与微流控平台兼容，并且可以很容易地适用于高效、高通量的HBV即时检测（POC）诊断。
