title: Molecular diagnosis of malaria in the field: development of a novel 1-step nucleic acid lateral flow immunoassay for the detection of all 4 human Plasmodium spp. and its evaluation in Mbita, Kenya.
abstract: Microscopy is frequently used for malaria diagnosis, but at low parasitemia, it becomes less sensitive and time consuming. Molecular tools allow for specific/sensitive diagnosis, but current formats, such as polymerase chain reaction (PCR) combined with gel electrophoresis and real-time PCR assays, are difficult to implement in resource-poor settings. Development of a simple, fast, sensitive, and specific detection system, nucleic acid lateral flow immunoassay (NALFIA) for amplified pan-Plasmodium PCR products, is described. The NALFIA lower detection limit is 0.3 to 3 parasites/microL, 10-fold more sensitive than gel electrophoresis analysis. Evaluating 650 clinically suspected malaria cases with the pan-Plasmodium assay under field conditions (rural Kenya) revealed that NALFIA detected more positives than microscopy (agreement, 95%; kappa value = 0.85), and there was an excellent agreement between gel electrophoresis and NALFIA (98.5%; kappa value = 0.96). In conclusion, NALFIA is more sensitive than microscopy and a good alternative to detect PCR products while circumventing using electricity or expensive equipment, making NALFIA the 1st step toward molecular field diagnosis.
authors.affiliation: Koninklijk Instituut voor de Tropen (KIT)/Royal Tropical Institute, KIT Biomedical Research, 1105 AZ Amsterdam, The Netherlands. p.mens@kit.nl
authors.forename: Petra F
authors.initials: PF
authors.lastname: Mens
authors.forename: Aart
authors.initials: A
authors.lastname: van Amerongen
authors.forename: Patrick
authors.initials: P
authors.lastname: Sawa
authors.forename: Piet A
authors.initials: PA
authors.lastname: Kager
authors.forename: Henk D F H
authors.initials: HD
authors.lastname: Schallig
mesh_terms: D000818:Animals; D002675:Child, Preschool; D016054:DNA, Protozoan / Q000032:analysis* / Q000235:genetics; D004587:Electrophoresis, Agar Gel; D005260:Female; D006801:Humans; D007118:Immunoassay; D007630:Kenya; D008288:Malaria / Q000175:diagnosis*; D008297:Male; D008853:Microscopy; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D010961:Plasmodium / Q000235:genetics / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity
chemical_list: D016054:DNA, Protozoan