High-throughput sequence typing of T-cell epitope polymorphisms in Plasmodium falciparum circumsporozoite protein

Mol Biochem Parasitol. 2000 Mar 5;106(2):273-82. doi: 10.1016/s0166-6851(99)00221-2.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alleles
  • Amino Acid Sequence
  • Animals
  • Antigens, Protozoan / genetics*
  • Base Sequence
  • DNA Primers / genetics
  • DNA, Protozoan / genetics
  • Epitopes / genetics
  • Gene Frequency
  • Humans
  • Malaria, Falciparum / parasitology
  • Molecular Sequence Data
  • Oligonucleotide Probes / genetics
  • Plasmodium falciparum / genetics*
  • Plasmodium falciparum / immunology*
  • Plasmodium falciparum / isolation & purification
  • Polymerase Chain Reaction
  • Polymorphism, Genetic*
  • Protozoan Proteins / genetics*
  • Protozoan Proteins / immunology*
  • T-Lymphocytes / immunology

Substances

  • Antigens, Protozoan
  • DNA Primers
  • DNA, Protozoan
  • Epitopes
  • Oligonucleotide Probes
  • Protozoan Proteins
  • circumsporozoite protein, Protozoan

Associated data

  • GENBANK/AF181833
  • GENBANK/AF181834
  • GENBANK/AF181835