Wang Chunfeng, Zhang Lianfeng, Shen Xuanmei
Digestive System Department, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, China.
Nucleosides Nucleotides Nucleic Acids. 2013;32(2):59-68. doi: 10.1080/15257770.2013.763976.
The object of this study was to develop a simple, rapid, specific, and highly sensitive method to detect HCV core antigen. A nucleic acid aptamer was designed with the high specificity and sensitivity in a nucleic acid lateral flow strip to compete with HCV core antigen and DNA probes. The lower detection limit of the test strip was calculated to be 10 pg/mL with the scanner and 100 pg/mL with naked eyes. Results showed that there were no cross-interactions with other proteins such as HCV NS3, E1/E2 antigens, HIV p24 antigens, or BSA proteins (HCV unrelated protein). When the viral load exceeded 10(4) copies/mL, the positive coincidence rates of ELISA and strip detection, when compared with the HCV RNA assay, were 98.44% and 97.28%, respectively. The results indicated that the ELISA detection and strip assay were in good agreement with the measured value. The results indicated that a nucleic acid lateral flow strip was a simple, rapid, specific, highly sensitive, and cost-effective field-based method for detecting HCV core antigen. The strip assay is an acceptable alternative to diagnose HCV core antigen and to investigate its epidemiology in clinical laboratories lacking specialized equipment and skills.
本研究的目的是开发一种简单、快速、特异且高度灵敏的检测丙型肝炎病毒（HCV）核心抗原的方法。设计了一种核酸适配体，其在核酸侧向流动条带中具有高特异性和灵敏度，可与HCV核心抗原及DNA探针竞争。经扫描仪检测，试纸条的最低检测限为10 pg/mL，肉眼检测为100 pg/mL。结果显示，该试纸条与其他蛋白质如HCV NS3、E1/E2抗原、HIV p24抗原或牛血清白蛋白（与HCV无关的蛋白质）无交叉反应。当病毒载量超过10⁴拷贝/mL时，与HCV RNA检测相比，ELISA和试纸条检测的阳性符合率分别为98.44%和97.28%。结果表明，ELISA检测和试纸条检测结果与测量值高度一致。结果表明，核酸侧向流动条带是一种简单、快速、特异、高度灵敏且经济高效的基于现场的HCV核心抗原检测方法。在缺乏专业设备和技术的临床实验室中，试纸条检测是诊断HCV核心抗原及其流行病学调查的一种可接受的替代方法。
