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Improving strategies for diagnosing infection by the four human Plasmodium species parasites is important as field-based epidemiologic and clinical studies focused on malaria become more ambitious. Expectations for malaria diagnostic assays include rapid processing with minimal expertise, very high specificity and sensitivity, and quantitative evaluation of parasitemia to be delivered at a very low cost. Toward fulfilling many of these expectations, we have developed a post–polymerase chain reaction (PCR)/ligase detection reaction–fluorescent microsphere assay (LDR-FMA). This assay, which uses Luminex® FlexMAPTM microspheres, provides simultaneous, semi-quantitative detection of infection by all four human malaria parasite species at a sensitivity and specificity equal to other PCR-based assays. In blinded studies using P. falciparum-infected blood from in vitro cultures, we identified infected and uninfected samples with 100% concordance. Additionally, in analyses of P. falciparum in vitro cultures and P. vivax-infected monkeys, comparisons between parasitemia and LDR-FMA signal intensity showed very strong positive correlations (r > 0.95). Application of this multiplex Plasmodium species LDR-FMA diagnostic assay will increase the speed, accuracy, and reliability of diagnosing human Plasmodium species infections in epidemiologic studies of complex malaria-endemic settings.
Received July 13, 2005. Accepted for publication August 29, 2005.
Acknowledgments: We thank Rajeev Mehlotra, Shannon Bruse, Jodi Thomson, JoAnn Sullivan, Kiet Dan Luc, and Hisashi Fujioka for technical assistance and advice during the course of this study. We also thank Sherry Dunbar, Christopher L. King, Ingrid Felger, and Ivo Mueller for helpful comments and criticisms during the preparation of the manuscript.
Financial support: This study was supported by grants from the National Institutes of Health (NIH) (AI-46919 and AI-52312). Brian T. Grimberg and Jennifer Cole-Tobian received support from NIH grant AI-07024.
* Address correspondence to Peter A. Zimmerman, Center for Global Health and Diseases, Case Western Reserve University School of Medicine, Wolstein Research Building, 4-125, Cleveland, OH 44106-7286. E-mail: paz{at}case.edu
Authors’ addresses: David T. McNamara, Laurin J. Kasehagen, Brian T. Grimberg, Jennifer Cole-Tobian, and Peter A. Zimmerman, Center for Global Health and Diseases, Case Western Reserve University School of Medicine, Wolstein Research Building, 4-125, Cleveland, OH 44106-7286. William E. Collins, Division of Parasitic Diseases, National Center for Infectious Diseases, Centers for Disease Control and Prevention, 4770 Buford Highway, Atlanta, GA 30341.
Reprint requests: Peter A. Zimmerman, Center for Global Health and Diseases, Case Western Reserve University School of Medicine, Wolstein Research Building, 4-125, Cleveland, OH 44106-7286, Telephone: 216-368-0508, Fax: 216-368-4825, E-mail: paz{at}case.edu.
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