Zhao Jiangtao, Feng Ruo
Department of Histology and Embryology, School of Basic Medical Sciences, Zhengzhou University, Zhengzhou, People's Republic of China.
Virus Genes. 2019 Feb;55(1):43-50. doi: 10.1007/s11262-018-1612-x. Epub 2018 Nov 13.
Zika virus (ZIKV) is a mosquito-borne flavivirus, which is a pathogen affecting humans in Africa, Asia, and America. It is necessary to detect ZIKV with a rapid and sensitive molecular method to guide timely treatment. In this study, a loop-mediated isothermal amplification (LAMP) assay was described, which is an attractive option as a fast, sensitive, and specific method for ZIKV detection using the NS5 protein coding region and the envelope protein (EP) coding region as target sequences. Two different techniques, a calcein/Mn complex chromogenic method and real-time turbidity monitoring, were employed. The specificity and sensitivity of the LAMP assay were determined. The assay's detection limit was 0.5 × 10 pmol/µl DNA for NS5 protein coding region and 1.12 × 10 pmol/µl DNA for E coding region, respectively, which is a 100-fold increase in sensitivity compared with real-time reverse transcription-polymerase chain reaction (RT-PCR) and conventional PCR. All 12 non-ZIKA respiratory pathogens tested were negative for LAMP detection, indicating the high specificity of the primers for ZIKV. In conclusion, a visual detection LAMP assay was developed, which could be a useful tool for primary quarantine purposes and clinical screening, especially in situations where resources are poor and in point-of-care tests.
寨卡病毒（ZIKV）是一种由蚊子传播的黄病毒，是一种在非洲、亚洲和美洲影响人类的病原体。有必要采用快速灵敏的分子方法检测寨卡病毒，以指导及时治疗。在本研究中，描述了一种环介导等温扩增（LAMP）检测方法，该方法作为一种快速、灵敏且特异的寨卡病毒检测方法颇具吸引力，它使用NS5蛋白编码区和包膜蛋白（EP）编码区作为靶序列。采用了两种不同技术，即钙黄绿素/锰复合物显色法和实时浊度监测法。测定了LAMP检测方法的特异性和灵敏度。该检测方法对NS5蛋白编码区的检测限为0.5×10 pmol/µl DNA，对E编码区的检测限为1.12×10 pmol/µl DNA，与实时逆转录聚合酶链反应（RT-PCR）和传统PCR相比，灵敏度提高了100倍。所检测的12种非寨卡病毒呼吸道病原体的LAMP检测均为阴性，表明寨卡病毒引物具有高度特异性。总之，开发了一种可视化检测LAMP检测方法，它可能是用于初步检疫目的和临床筛查的有用工具，特别是在资源匮乏的情况下和即时检测中。
