Han Qiuxue, Zhang Shengnan, Liu Dongping, Yan Feihu, Wang Hualei, Huang Pei, Bi Jinhao, Jin Hongli, Feng Na, Cao Zengguo, Gao Yuwei, Chi Hang, Yang Songtao, Zhao Yongkun, Xia Xianzhu
Institute of Laboratory Animal Science, Chinese Academy of Medical Sciences (CAMS) and Comparative Medicine Center, Peking Union Medical College (PUMC), Beijing, China.
Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary Medicine, Academy of Military Medical Sciences, Changchun, China.
Front Microbiol. 2020 Nov 13;11:590732. doi: 10.3389/fmicb.2020.590732. eCollection 2020.
Rift Valley fever (RVF) is a severe infectious disease, which can through mosquito bites, direct contact and aerosol transmission infect sheep, goats, people, camels, cattle, buffaloes, and so on. In this paper, a conserved region of the S RNA segment of Rift Valley fever virus (RVFV) ZH501 strain was used as target sequence. The RVFV RT-LAMP-VF assay was successfully established combined reverse transcription-loop-mediated isothermal amplification with a vertical flow visualization strip. The detection limit is up to 1.94 × 10 copies/μl of synthesized RVFV-RNA. RNA extracted from cell culture of an inactivated RVFV-BJ01 strain was also used as templates, and the detection limit is 1.83 × 10 copies/μl. In addition, there was no cross-reactivity with other viruses that can cause similar fever symptoms. The RVFV-LAMP-VF assay exhibited very high levels of diagnostic sensitivity, which had 100-fold more sensitive than RVFV real-time RT-PCR assay. Accordingly, the RVFV RT-LAMP-VF assay developed in this study is suitable for the rapid and sensitive diagnosis of RVFV without specialized equipment and can rapidly complete detection within 60 min, and the results are visible by vertical flow visualization strip within 5 min.
裂谷热（RVF）是一种严重的传染病，可通过蚊虫叮咬、直接接触和气溶胶传播感染绵羊、山羊、人、骆驼、牛、水牛等。本文以裂谷热病毒（RVFV）ZH501株S RNA片段的保守区为靶序列，成功建立了将逆转录-环介导等温扩增与垂直流可视化条带相结合的RVFV RT-LAMP-VF检测方法。检测限高达1.94×10拷贝/μl合成的RVFV-RNA。从灭活的RVFV-BJ01株细胞培养物中提取的RNA也用作模板，检测限为1.83×10拷贝/μl。此外，与其他可引起类似发热症状的病毒无交叉反应。RVFV-LAMP-VF检测方法具有很高的诊断灵敏度，比RVFV实时RT-PCR检测方法灵敏100倍。因此，本研究建立的RVFV RT-LAMP-VF检测方法适用于在无需专门设备的情况下对RVFV进行快速灵敏诊断，可在60分钟内快速完成检测，结果在5分钟内通过垂直流可视化条带可见。
