A polymerase chain reaction/ligase detection reaction fluorescent microsphere assay to determine Plasmodium falciparum MSP-119 haplotypes.
The merozoite surface protein-1 (MSP-1) is a blood stage antigen currently being tested as a vaccine against Plasmodium falciparum malaria. Determining the MSP-1(19) haplotype(s) present during infection is essential for assessments of MSP-1 vaccine efficacy and studies of protective immunity in human populations. The C-terminal fragment (MSP-1(19)) has four predominant haplotypes based on point mutations resulting in non-synonymous amino acid changes: E-TSR (PNG-MAD20 type), E-KNG (Uganda-PA type), Q-KNG (Wellcome type), and Q-TSR (Indo type). Current techniques using direct DNA sequencing are laborious and expensive. We present an MSP-1(19) allele-specific polymerase chain reaction (PCR)/ligase detection reaction-fluorescent microsphere assay (LDR-FMA) that allows simultaneous detection of the four predominant MSP-1(19) haplotypes with a sensitivity and specificity comparable with other molecular methods and a semi-quantitative determination of haplotype contribution in mixed infections. Application of this method is an inexpensive, accurate, and high-throughput alternative to distinguish the predominant MSP-1(19) haplotypes in epidemiologic studies.
Dent
Arlene E
AE
Center for Global Health and Diseases, Case Western Reserve University School of Medicine, 10900 Euclid Avenue, Cleveland, OH 44106, USA. arlene.dent@case.edu
Yohn
Christopher T
CT
Zimmerman
Peter A
PA
Vulule
John
J
Kazura
James W
JW
Moormann
Ann M
AM
D000328:Adult; D000818:Animals; D002648:Child; D016054:DNA, Protozoan / Q000737:chemistry / Q000235:genetics; D005456:Fluorescent Dyes; D006239:Haplotypes; D006801:Humans; D007630:Kenya; D017780:Malaria Vaccines / Q000276:immunology*; D016778:Malaria, Falciparum / Q000453:epidemiology / Q000469:parasitology* / Q000517:prevention & control; D020066:Merozoite Surface Protein 1 / Q000235:genetics / Q000276:immunology*; D008863:Microspheres; D010963:Plasmodium falciparum / Q000235:genetics / Q000276:immunology*; D016133:Polymerase Chain Reaction / Q000379:methods*; D015995:Prevalence; D012337:RNA, Ribosomal, 18S / Q000737:chemistry / Q000235:genetics; D012680:Sensitivity and Specificity
D016054:DNA, Protozoan; D005456:Fluorescent Dyes; D017780:Malaria Vaccines; D020066:Merozoite Surface Protein 1; D012337:RNA, Ribosomal, 18S
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