Xia Xiaoming, Yu Yongxin, Hu Linghao, Weidmann Manfred, Pan Yingjie, Yan Shuling, Wang Yongjie
Laboratory of Quality and Safety Risk Assessment for Aquatic Products on Storage and Preservation, Ministry of Agriculture, Shanghai, China.
Arch Virol. 2015 Apr;160(4):987-94. doi: 10.1007/s00705-015-2357-7. Epub 2015 Feb 6.
Infectious hypodermal and hematopoietic necrosis virus (IHHNV) causes mortality or runt deformity syndrome in penaeid shrimps and is responsible for significant economic losses in the shrimp aquaculture industry. Here, we describe a novel real-time isothermal recombinase polymerase amplification (RPA) assay developed for IHHNV detection. Using IHHNV plasmid standards and DNA samples from a variety of organisms, we evaluated the ability of the IHHNV-RPA assay to detect IHHNV based on analysis of its sensitivity, specificity, rapidity, and reproducibility. Probit analysis of eight independent experimental replicates indicated satisfactory performance of the RPA assay, which is sufficiently sensitive to detect as few as 4 copies of the IHHNV genome within 7 min at 39 °C with 95 % reliability. Therefore, this rapid RPA method has great potential for applications, either in field use or as a point of care diagnostic technique.
传染性皮下和造血组织坏死病毒（IHHNV）可导致对虾出现死亡或侏儒畸形综合征，并给对虾养殖业造成重大经济损失。在此，我们描述了一种用于检测IHHNV的新型实时等温重组酶聚合酶扩增（RPA）检测方法。使用IHHNV质粒标准品和来自多种生物的DNA样本，我们基于对其灵敏度、特异性、快速性和可重复性的分析，评估了IHHNV-RPA检测方法检测IHHNV的能力。对八个独立实验复制品的概率分析表明RPA检测方法性能良好，该方法在39℃下7分钟内能够灵敏地检测到低至4个拷贝的IHHNV基因组，可靠性达95%。因此，这种快速的RPA方法无论是在现场使用还是作为即时诊断技术都具有巨大的应用潜力。
