A high-throughput method for quantifying alleles and haplotypes of the malaria vaccine candidate Plasmodium falciparum merozoite surface protein-1 19 kDa.

Malaria Journal
Shannon L TakalaChristopher V Plowe

Abstract

Malaria vaccine efficacy may be compromised if the frequency of non-target alleles increases following vaccination with a genetically polymorphic target. Methods are needed to monitor genetic diversity in polymorphic vaccine antigens, but determining which genetic variants of such antigens are present in infected individuals is complicated by the frequent occurrence of mixed infections. Pyrosequencing was used to determine allele frequencies at each of six single nucleotide polymorphisms in the Plasmodium falciparum blood-stage vaccine antigen merozoite surface protein 1 19 kDa (MSP-119) in field samples from a vaccine-testing site in Mali. Mixtures of MSP-119 clones were created to validate a haplotype-estimating algorithm that uses maximum likelihood methods to determine the most probable combination of haplotypes given the allele frequencies for an infection and the haplotypes known to be circulating in the population. Fourteen unique MSP-119 haplotypes were identified among 351 genotyped infections. After adjustment to a standard curve, Pyrosequencing provided accurate and precise estimates of allele frequencies in mixed infections. The haplotype-estimating algorithm provided accurate estimates of haplotypes in mixed infect...Continue Reading

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Citations

Sep 20, 2008·Current Genetics·Karen Hayton, Xin-Zhuan Su
Feb 1, 2013·Molecular Biology and Evolution·Darren KessnerJohn Novembre
Nov 25, 2006·PLoS Clinical Trials·Mahamadou A TheraUNKNOWN Mali FMP1 Working Group
Mar 16, 2007·PLoS Medicine·Shannon L TakalaChristopher V Plowe
Feb 13, 2015·The Journal of Infectious Diseases·Elena ArtimovichShannon Takala-Harrison
Mar 9, 2007·Molecular and Biochemical Parasitology·Jeffrey J BabonAnthony A Holder
Apr 24, 2013·Statistics in Medicine·Leonore WiggerVolker Roth
Aug 14, 2013·Molecular Ecology Resources·Sabine Giessler, Justyna Wolinska

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Methods Mentioned

BETA
PCR
Assay

Software Mentioned

Pyrosequencing Assay Design
R Foundation for Statistical Computing
R
Pyrosequencing

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