High-throughput sequence typing of T-cell epitope polymorphisms in Plasmodium falciparum circumsporozoite protein.
We report a method for typing polymorphisms at the T-cell epitopes within the Th2R and Th3R regions of the Plasmodium falciparum circumsporozoite protein (CSP). This method combines the use of PCR and sequence specific oligonucleotide probes (PCR-SSOP), and allows the identification of single nucleotide polymorphisms in these epitope regions. PCR-SSOP is a robust and a high-throughput sequence typing technique which has the same specificity and fidelity as direct sequencing. This method has been developed specifically for the assessment of the protective efficacy of RTS,S/SBAS2 vaccine against the 3D7 strain of P. falciparum (RTS,S/SBAS2 vaccine contains a part of the 3D7 CSP protein) in a phase IIb trial in Gambia which has been completed recently. PCR-SSOP could be used to determine the allelic frequencies of other parasite antigens and their geographical distribution.
Alloueche
A
A
Department of Infectious and Tropical Diseases, London School of Hygiene and Tropical Medicine, Keppel Street, London, UK. ali.alloueche@lshtm.ac.uk
Silveira
H
H
Conway
D J
DJ
Bojang
K
K
Doherty
T
T
Cohen
J
J
Pinder
M
M
Greenwood
B M
BM
D000483:Alleles; D000595:Amino Acid Sequence; D000818:Animals; D000953:Antigens, Protozoan / Q000235:genetics*; D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics; D016054:DNA, Protozoan / Q000235:genetics; D000939:Epitopes / Q000235:genetics; D005787:Gene Frequency; D006801:Humans; D016778:Malaria, Falciparum / Q000469:parasitology; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes / Q000235:genetics; D010963:Plasmodium falciparum / Q000235:genetics* / Q000276:immunology* / Q000302:isolation & purification; D016133:Polymerase Chain Reaction; D011110:Polymorphism, Genetic; D015800:Protozoan Proteins / Q000235:genetics* / Q000276:immunology*; D013601:T-Lymphocytes / Q000276:immunology
D000953:Antigens, Protozoan; D017931:DNA Primers; D016054:DNA, Protozoan; D000939:Epitopes; D015345:Oligonucleotide Probes; D015800:Protozoan Proteins; C041595:circumsporozoite protein, Protozoan