Diagnostic systems that can deliver highly specific and sensitive detection of hepatitis A virus (HAV) in food and water are of particular interest in many fields including food safety, biosecurity and control of outbreaks. Our aim was the development of an electrochemical method based on DNA hybridization to detect HAV. A ssDNA probe specific for HAV (capture probe) was designed and tested on DNAs from various viral and bacterial samples using Nested-Reverse Transcription Polymerase Chain Reaction (nRT-PCR). To develop the electrochemical device, a disposable gold electrode was functionalized with the specific capture probe and tested on complementary ssDNA and on HAV cDNA. The DNA hybridization on the electrode was measured through the monitoring of the oxidative peak potential of the indicator tripropylamine by cyclic voltammetry. To prevent non-specific binding the gold surface was treated with 3% BSA before detection. High resolution atomic force microscopy (AFM) confirmed the efficiency of electrode functionalization and on-electrode hybridization. The proposed device showed a limit of detection of 0.65 pM for the complementary ssDNA and 6.94 fg/µL for viral cDNA. For a comparison, nRT-PCR quantified the target HAV cDNA with a limit of detection of 6.4 fg/µL. The DNA-sensor developed can be adapted to a portable format to be adopted as an easy-to- use and low cost method for screening HAV in contaminated food and water. In addition, it can be useful for rapid control of HAV infections as it takes only a few minutes to provide the results.

在食品安全，生物安全和疫情控制等许多领域中，可以对食物和水中的甲型肝炎病毒（HAV）进行高度特异性和灵敏检测的诊断系统尤为重要。我们的目标是开发一种基于DNA杂交检测HAV的电化学方法。设计了对HAV特异的ssDNA探针（捕获探针），并使用嵌套逆转录聚合酶链反应（nRT-PCR）对来自各种病毒和细菌样本的DNA进行了测试。为了开发电化学装置，将一次性金电极与特异性捕获探针进行功能化，并在互补ssDNA和HAV cDNA上进行测试。通过用循环伏安法监测指示剂三丙胺的氧化峰电位来测量电极上的DNA杂交。为了防止非特异性结合，在检测之前，将金表面用3％BSA处理。高分辨率原子力显微镜（AFM）证实了电极功能化和电极上杂交的效率。拟议的设备显示互补ssDNA的检测限为0.65 pM，病毒cDNA的检测限为6.94 fg / µL。为了进行比较，nRT-PCR对目标HAV cDNA进行了定量，检测限为6.4 fg / µL。所开发的DNA传感器可适应便携式格式，以作为一种易于使用且低成本的方法来筛查受污染的食物和水中的HAV。此外，它对快速控制HAV感染很有用，因为只需几分钟即可提供结果。高分辨率原子力显微镜（AFM）证实了电极功能化和电极上杂交的效率。拟议的设备显示互补ssDNA的检测限为0.65 pM，病毒cDNA的检测限为6.94 fg / µL。为了进行比较，nRT-PCR对目标HAV cDNA进行了定量，检测限为6.4 fg / µL。所开发的DNA传感器可适应便携式格式，以作为一种易于使用且低成本的方法来筛查受污染的食物和水中的HAV。此外，它对快速控制HAV感染很有用，因为只需几分钟即可提供结果。高分辨率原子力显微镜（AFM）证实了电极功能化和电极上杂交的效率。拟议的设备显示互补ssDNA的检测限为0.65 pM，病毒cDNA的检测限为6.94 fg / µL。为了进行比较，nRT-PCR对目标HAV cDNA进行了定量，检出限为6.4 fg / µL。所开发的DNA传感器可适应便携式格式，以作为一种易于使用且低成本的方法来筛查受污染的食物和水中的HAV。此外，它对快速控制HAV感染很有用，因为只需几分钟即可提供结果。nRT-PCR对目标HAV cDNA进行定量，检出限为6.4 fg / µL。所开发的DNA传感器可适应便携式格式，以作为一种易于使用且低成本的方法来筛查受污染的食物和水中的HAV。此外，它对快速控制HAV感染很有用，因为只需几分钟即可提供结果。nRT-PCR对目标HAV cDNA进行定量，检出限为6.4 fg / µL。所开发的DNA传感器可适应便携式格式，以作为一种易于使用且低成本的方法来筛查受污染的食物和水中的HAV。此外，它对快速控制HAV感染很有用，因为只需几分钟即可提供结果。