Department of Quantitative Health Sciences; Department of Pediatrics
Adult; Animals; Child; DNA, Protozoan; Fluorescent Dyes; Haplotypes; Humans; Kenya; Malaria Vaccines; Malaria, Falciparum; Merozoite Surface Protein 1; Microspheres; Plasmodium falciparum; Polymerase Chain Reaction; Prevalence; RNA, Ribosomal, 18S; Sensitivity and Specificity
Biostatistics | Epidemiology | Health Services Research | Immunology and Infectious Disease | Pediatrics
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.
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Copyright © 2007 by The American Society of Tropical Medicine and Hygiene.
Am J Trop Med Hyg. 2007 Aug;77(2):250-5. Link to article on publisher's site
The American journal of tropical medicine and hygiene
Dent, Arlene E.; Yohn, Christopher T.; Zimmerman, Peter A.; Vulule, John M.; Kazura, James W.; and Moormann, Ann M., "A polymerase chain reaction/ligase detection reaction fluorescent microsphere assay to determine Plasmodium falciparum MSP-119 haplotypes" (2007). Quantitative Health Sciences Publications and Presentations. 399.