Abstract
Purpose: To develop a rapid detection method for Rift Valley fever virus (RVFV) diagnosis.Methods: According to the reference sequences of RVFV published in GenBank, nine overlapping polymerase chain reaction (PCR) primers and four specific reverse transcription loop-mediated isothermal amplification (RT-LAMP) primers were designed using DNAStar and LAMP primer design software, respectively. Based on the synthesis of a conserved part of the RVFV S segment gene sequence using overlapping PCR, RT-LAMP assay was first established and evaluated after a series of tests, including, optimization of reaction conditions, and sensitivity and specificity tests.Result: A target RVFV S segment gene fragment of 288 bp was synthesised. The optimal reaction conditions for RT-LAMP assay were 63 °C for 45 min: the assay has a specific ladder-like pattern of amplification bands from about 120 bp. The lowest target gene copy number of RT-LAMP for RVFV detection was 70 copies. The assay showed good specificity as only the synthesised target RVFV gene was amplified with no amplification for the detection of Peste des petits ruminants virus, Epidemic encephalitis B virus, E. coli, Pasteurella multocida, or Salmonella.Conclusion: This study provides a rapid, sensitive, specific RT-LAMP method for RVFV detection.Keywords: Rift valley fever virus, Overlapping polymerase chain reaction, Reverse transcription loopmediated isothermal amplification, Rapid diagnosis test
Highlights
Rift Valley fever (RVF), resulting from the RVF virus (RVFV), is an acute and febrile infection of both ruminants and humans [1]
RVFV is a typosis in some African countries and areas, but has spread to areas of the Mediterranean coast and Middle East, showing its potential to spread to areas outside of Africa [1]
As these mosquitoes are distributed around the world, RVFV is prone to spread to Europe, America, and other non-epidemic areas [5,8,9,10]
Summary
Rift Valley fever (RVF), resulting from the RVF virus (RVFV), is an acute and febrile infection of both ruminants and humans [1]. It was first found in the Great Rift Valley in Kenya in 1930 [1]. RVFV shows common resistance to the outside environment, it presents strong transmission capability. It can be transmitted by aerosol, touch, or consumption of infected animal tissue, and vertically via the placenta, as well as by mosquito bites [5,6,7]. Due to the fact that there are no efficient vaccines for humans or animals [5], controlling and conducting research on RVF have becoming more urgent and significant
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