A polymerase chain reaction/ligase detection reaction fluorescent microsphere assay to determine Plasmodium falciparum MSP-119 haplotypes
Dent, Arlene E. ; Yohn, Christopher T. ; Zimmerman, Peter A. ; Vulule, John M. ; Kazura, James W. ; Moormann, Ann M.
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Student Authors
Faculty Advisor
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UMass Chan Affiliations
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Keywords
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
Subject Area
Embargo Expiration Date
Link to Full Text
Abstract
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.
Source
Am J Trop Med Hyg. 2007 Aug;77(2):250-5. Link to article on publisher's site