Using multiplex amplicon pcr technology to efficiently and timely generate rift valley fever virus sequence data for genomic surveillance

Simple item page
dc.contributor.authorJuma, John
dc.contributor.authorChristoffels, Alan
dc.contributor.authorRoesel, Kristina
dc.date.accessioned2023-04-12T08:14:39Z
dc.date.available2023-04-12T08:14:39Z
dc.date.issued2023
dc.description.abstractRift Valley fever (RVF) is a febrile vector-borne disease endemic in Africa and continues to spread in new territories. It is a climate-sensitive disease mostly triggered by abnormal rainfall patterns. The disease is associated with high mortality and morbidity in both humans and livestock. RVF is caused by the Rift Valley fever virus (RVFV) of the genus Phlebovirus in the family Phenuiviridae. It is a tripartite RNA virus with three genomic segments: small (S), medium (M) and large (L). Pathogen genomic sequencing is becoming a routine procedure and a powerful tool for understanding the evolutionary dynamics of infectious organisms, including viruses. Inspired by the utility of amplicon-based sequencing demonstrated in severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) and Ebola, Zika and West Nile viruses, we report an RVFV sample preparation based on amplicon multiplex polymerase chain reaction (amPCR) for template enrichment and reduction of background host contamination. The technology can be implemented rapidly to characterize and genotype RVFV during outbreaks in a near-real-time manner. To achieve this, we designed 74 multiplex primer sets covering the entire RVFV genome to specifically amplify the nucleic acid of RVFV in clinical samples from an animal tissue. Using this approach, we demonstrate achieving complete RVFV genome coverage even from samples containing a relatively low viral load. We report the first primer scheme approach of generating multiplex primer sets for a tripartite virus which can be replicated for other segmented viruses.en_US
dc.identifier.citationJuma, J., Konongoi, S. L., Nsengimana, I., Mwangi, R., Akoko, J., Nyamota, R., Muli, C., et al. (2023). Using Multiplex Amplicon PCR Technology to Efficiently and Timely Generate Rift Valley Fever Virus Sequence Data for Genomic Surveillance. Viruses, 15(2), 477. MDPI AG. Retrieved from http://dx.doi.org/10.3390/v15020477en_US
dc.identifier.urihttps://doi.org/10.3390/v15020477
dc.identifier.urihttp://hdl.handle.net/10566/8747
dc.language.isoenen_US
dc.publisherMDPIen_US
dc.subjectampliconen_US
dc.subjectmultiplex PCRen_US
dc.subjectenrichmenten_US
dc.subjectcultureen_US
dc.subjectgenomeen_US
dc.subjectRift Valley feveren_US
dc.titleUsing multiplex amplicon pcr technology to efficiently and timely generate rift valley fever virus sequence data for genomic surveillanceen_US
dc.typeArticleen_US

Files

Original bundle
Now showing 1 - 1 of 1
Thumbnail Image
Name:
Juma J_Using Multiplex Amplicon PCR Technology to Efficiently and Timely Generate Rift Valley Fever Virus Sequence Data for Genomic Surveillance_2023.pdf
Size:
686.65 KB
Format:
Adobe Portable Document Format
Description:
Download
License bundle
Now showing 1 - 1 of 1
No Thumbnail Available
Name:
license.txt
Size:
1.71 KB
Format:
Item-specific license agreed upon to submission
Description:
Download

Collections

Research Articles (Biotechnology)