title: Application of Droplet Digital PCR to Validate Rift Valley Fever Vaccines.
abstract: Droplet Digital™ polymerase chain reaction (ddPCR™) is a promising technique that quantitates the absolute concentration of nucleic acids in a given sample. This technique utilizes water-in-oil emulsion technology, a system developed by Bio-Rad Laboratories that partitions a single sample into thousands of nanoliter-sized droplets and counts nucleic acid molecules encapsulated in each individual particle as one PCR reaction. This chapter discusses the applications and methodologies of ddPCR for development of Rift Valley fever (RVF) vaccine, using an example that measures RNA copy numbers of a live-attenuated MP-12 vaccine from virus stocks, infected cells, or animal blood. We also discuss how ddPCR detects a reversion mutant of MP-12 from virus stocks accurately. The use of ddPCR improves the quality control of live-attenuated vaccines in the seed lot systems.
authors.affiliation: Department of Pathology, The University of Texas Medical Branch at Galveston, 301 University Blvd., Galveston, TX, 77555, USA.
authors.forename: Hoai J
authors.initials: HJ
authors.lastname: Ly
authors.affiliation: Department of Pathology, The University of Texas Medical Branch at Galveston, 301 University Blvd., Galveston, TX, 77555, USA.
authors.forename: Nandadeva
authors.initials: N
authors.lastname: Lokugamage
authors.affiliation: Department of Pathology, The University of Texas Medical Branch at Galveston, 301 University Blvd., Galveston, TX, 77555, USA. teikegam@utmb.edu.
authors.forename: Tetsuro
authors.initials: T
authors.lastname: Ikegami
mesh_terms: D016133:Polymerase Chain Reaction / Q000379:methods*; D012367:RNA, Viral / Q000235:genetics; D012296:Rift Valley fever virus / Q000276:immunology*; D014613:Vaccines, Attenuated / Q000276:immunology*; D014765:Viral Vaccines / Q000276:immunology*
chemical_list: D012367:RNA, Viral; D014613:Vaccines, Attenuated; D014765:Viral Vaccines
keywords: Copy number validation; Droplet digital PCR; MP-12 vaccine; Rift Valley fever; Variant RNA detection