Abstract
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 × 100 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 × 103 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.
Highlights
Rift Valley fever virus (RVFV) is an arthropod-borne, zoonotic virus that is a significant threat to domestic ruminants and humans (Elliott, 1997)
Synthetic RVFV-RNA samples were used as the template to optimize the amplification conditions for the RVFV RT-loop-mediated isothermal amplification (LAMP)-VF assay
The lowest standard template copy value, 0.935 × 102 copies/μl, was detected at 63°C, and so 63°C was considered as the optimum amplification temperature for the RVFV RT-LAMP-VF assay
Summary
Rift Valley fever virus (RVFV) is an arthropod-borne, zoonotic virus (genus Phlebovirus, family Phenuivirudae) that is a significant threat to domestic ruminants and humans (Elliott, 1997). A RVFV-4seGFP based VNT can be completed in just 48 h (Wichgers Schreur et al, 2017) Since it requires live virus and can be done only in biocontainment facilities, the virus production requires biocontainment facilities to limit the risk of exposure of laboratory personnel to infection (Paweska et al, 2003). Sensitive PCR assays for the detection and quantification of RVFV have been reported, including reverse transcriptase PCR (Garcia et al, 2001; Sall et al, 2002) and real-time RT-PCR (Drosten et al, 2002; Tercero et al, 2019), which require sophisticated equipped laboratories and might be beyond the resources and capabilities of many developing countries. There is still a lack of a sensitive, intuitive reading method, which is more suitable for on-site testing
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