Application of droplet digital PCR to validate rift valley fever vaccines

Hoai J. Ly, Nandadeva Lokugamage, Tetsuro Ikegami

Research output: Chapter in Book/Report/Conference proceedingChapter

2 Scopus citations

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.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages207-220
Number of pages14
Volume1403
DOIs
StatePublished - Apr 1 2016

Publication series

NameMethods in Molecular Biology
Volume1403
ISSN (Print)10643745

Keywords

  • Copy number validation
  • Droplet digital PCR
  • MP-12 vaccine
  • Rift valley fever
  • Variant RNA detection

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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  • Cite this

    Ly, H. J., Lokugamage, N., & Ikegami, T. (2016). Application of droplet digital PCR to validate rift valley fever vaccines. In Methods in Molecular Biology (Vol. 1403, pp. 207-220). (Methods in Molecular Biology; Vol. 1403). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-3387-7_10