Evaluation of a new rapid molecular diagnostic system for Plasmodium falciparum combined with DNA filter paper, loop-mediated isothermal amplification, and melting curve analysis.
Falciparum malaria is a fatal infection without immediate diagnosability or treatment. There are shortages of clinicians and examiners skilled in the treatment of malaria in non-endemic countries, including Japan. This study was performed to evaluate a novel rapid molecular diagnostic system consisting of loop-mediated isothermal amplification (LAMP) combined with DNA filter paper (FTA card) and melting curve analysis. Combining LAMP with melting curve analysis enabled diagnosis of Plasmodium falciparum more accurately with relative ease. FTA cards could be used to clarify problems regarding storage, infectivity, and transportation. The LAMP assay was carried out at a constant temperature of 63 degrees C for 90 min. The diagnostic system (malaria-LAMP) accurately diagnosed malaria (47 samples from Thailand and 50 from Zimbabwe) with 97.8% sensitivity and 85.7% specificity as compared with microscopic methods, indicating the usefulness of this combined system.
Yamamura
Mariko
M
Department of Internal Medicine, Teikyo University School of Medicine, Tokyo 173-0003, Japan. yamariko829@excite.co.jp
Makimura
Koichi
K
Ota
Yasuo
Y
D000818:Animals; D016054:DNA, Protozoan / Q000032:analysis* / Q000235:genetics; D005374:Filtration; D006801:Humans; D007564:Japan; D016778:Malaria, Falciparum / Q000097:blood / Q000175:diagnosis* / Q000469:parasitology; D008969:Molecular Sequence Data; D010963:Plasmodium falciparum / Q000235:genetics / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012337:RNA, Ribosomal, 18S / Q000032:analysis* / Q000235:genetics; D012680:Sensitivity and Specificity; D013048:Specimen Handling / Q000379:methods*; D044366:Transition Temperature
D016054:DNA, Protozoan; D012337:RNA, Ribosomal, 18S