Comparison of RT-qPCR and RT-ddPCR with Rift valley fever virus (RVFV) RNA

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作者
Changwoo Park
Dongju Park
Zohaib Ul Hassan
Sang Ho Choi
Seil Kim
机构
[1] Korea Research Institute of Standards and Science,Microbiological Analysis Team, Group for Biometrology
[2] Korea Research Institute of Chemical Technology,Convergent Research Center for Emerging Virus Infection
[3] Seoul National University,Department of Agricultural Biotechnology
[4] Theragen Bio Co.,Department of Bio
[5] Ltd.,Analysis Science
[6] University of Science & Technology,Center for Food and Bioconvergence
[7] Seoul National University,undefined
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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.
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