Microbiological Analysis Team, Group for Biometrology, Korea Research Institute of Standards and Science, Daejeon, 34113, Republic of Korea.
Convergent Research Center for Emerging Virus Infection, Korea Research Institute of Chemical Technology, Daejeon, 34114, Republic of Korea.
Sci Rep. 2023 Feb 22;13(1):3085. doi: 10.1038/s41598-023-29023-y.
Rift valley fever (RVF) is an important zoonotic disease caused by the Rift valley fever virus (RVFV) which can affect ruminants and humans. In this study, a comparison was done of the reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and reverse transcription-droplet digital PCR (RT-ddPCR) assays with synthesized RVFV RNA, cultured viral RNA, and mock clinical RVFV RNA samples. The genomic segments (L, M, and S) of three RVFV strains (BIME01, Kenya56, and ZH548) were synthesized and used as templates for in vitro transcription (IVT). Both the RT-qPCR and RT-ddPCR assays for RVFV did not react with any of the negative reference viral genomes. Thus, both the RT-qPCR and RT-ddPCR assays are specific to RVFV. The comparison of both the RT-qPCR and RT-ddPCR assays with serially diluted templates showed that the LoD of both assays are similar, and a concordant of the results was observed. The LoD of both assays reached the practical measurable minimum concentration. Taken altogether, the sensitivity of the RT-qPCR and RT-ddPCR assays is similar, and the material measured by RT-ddPCR can be used as a reference material for RT-qPCR.
裂谷热（RVF）是一种由裂谷热病毒（RVFV）引起的重要人畜共患病，可影响反刍动物和人类。在这项研究中，对合成的 RVFV RNA、培养的病毒 RNA 和模拟临床 RVFV RNA 样本的逆转录定量聚合酶链反应（RT-qPCR）和逆转录-数字液滴 PCR（RT-ddPCR）检测进行了比较。三种 RVFV 株（BIME01、肯尼亚 56 和 ZH548）的基因组片段（L、M 和 S）被合成，并用作体外转录（IVT）的模板。用于 RVFV 的 RT-qPCR 和 RT-ddPCR 检测均未与任何阴性参考病毒基因组发生反应。因此，RT-qPCR 和 RT-ddPCR 检测均对 RVFV 具有特异性。用连续稀释模板对两种 RT-qPCR 和 RT-ddPCR 检测的比较表明，两种检测的 LoD 相似，并且观察到结果一致。两种检测的 LoD 均达到了实际可测量的最小浓度。总的来说，RT-qPCR 和 RT-ddPCR 检测的灵敏度相似，并且 RT-ddPCR 测量的材料可用作 RT-qPCR 的参考材料。
