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病毒载量测量对于人类免疫缺陷病毒(HIV)阳性患者长期临床护理来说是一个必不可少的工具。然而,考虑到病毒载量测量所需的仪器体积、成本和操作的复杂性,在医疗基础设施较差的偏远地区(尤其是在被HIV感染人群比例较高的地区)普及标准的病毒载量测量仪器是较为困难的。为提高该检测方法的普及性,人们已经开始开发可以进行即时检测的病毒载量检测平台,然而尚没有解决办法能够同时满足低成本、便携、易于操作等多种实际要求。本文通过运用微流体和微型硅晶片平台,对经过最低程度处理的含有HIV的全血样本进行了逆转录环介导等温扩增(RT-LAMP),并利用智能手机进行了荧光检测。集成实验检测结果表明,一滴约60 n L的反应液滴中仅有的3个病毒依然可以通过RT-LAMP技术被检测到,这相当于每微升全血样品中只有670个病毒。该技术在数字化RT-LAMP方法上具有重要意义,扩展该技术能够实现对HIV阳性患者在临床护理中采集指血进行病毒载量检测。研究结果显示,病毒载量检测过程所需的各个步骤,从血滴的准备到RT-LAMP反应的成像,都可以集成为晶片实验并且可以和移动设备兼容。
Viral load measurement is an essential tool for long-term clinical care in HIV-positive patients. However, given the volume, cost and operational complexity of instrumentation required for viral load measurement, standard viral load is widespread in remote areas where medical infrastructure is poor, especially in areas with a high HIV-infected population Measuring instruments is more difficult. In order to improve the popularity of this detection method, a viral load detection platform that can perform real-time detection has been developed. However, there are still no solutions to meet the practical requirements of low cost, portability and ease of operation. In this work, RT-LAMP was performed on a minimally-processed HIV-containing whole blood sample using microfluidics and micro-silicon wafer platforms, and fluorescence detection was performed using a smartphone. The integrated laboratory test results showed that only 3 of a drop of approximately 60 nL of reaction droplets could still be detected by RT-LAMP technology, which corresponds to only 670 viruses per microliter of whole blood samples. This technique is of great importance in the digital RT-LAMP method. The expansion of this technique enables the detection of viral load in HIV-positive patients by finger-picking blood from clinical care. The results show that the various steps required for the viral load testing process, from preparation of blood drops to imaging of RT-LAMP responses, can be integrated into wafer experiments and are compatible with mobile devices.