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Journal of Clinical Microbiology, June 2004, p. 2403-2410, Vol. 42, No. 6
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.6.2403-2410.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Development of a Multiplex PCR-Ligase Detection Reaction Assay for Diagnosis of Infection by the Four Parasite Species Causing Malaria in Humans

David T. McNamara, Jodi M. Thomson, Laurin J. Kasehagen, and Peter A. Zimmerman*

The Center for Global Health and Diseases, Case Western Reserve University School of Medicine, Cleveland, Ohio 44106-7286

Received 22 August 2003/ Returned for modification 28 October 2003/ Accepted 11 January 2004

The diagnosis of infections caused by Plasmodium species is critical for understanding the nature of malarial disease, treatment efficacy, malaria control, and public health. The demands of field-based epidemiological studies of malaria will require faster and more sensitive diagnostic methods as new antimalarial drugs and vaccines are explored. We have developed a multiplex PCR-ligase detection reaction (LDR) assay that allows the simultaneous diagnosis of infection by all four parasite species causing malaria in humans. This assay exhibits sensitivity and specificity equal to those of other PCR-based assays, identifying all four human malaria parasite species at levels of parasitemias equal to 1 parasitized erythrocyte/µl of blood. The multiplex PCR-LDR assay goes beyond other PCR-based assays by reducing technical procedures and by detecting intraindividual differences in species-specific levels of parasitemia. Application of the multiplex PCR-LDR assay will provide the sensitivity and specificity expected of PCR-based diagnostic assays and will contribute new insight regarding relationships between the human malaria parasite species and the human host in future epidemiological studies.

* Corresponding author. Mailing address: The Center for Global Health and Diseases, Case Western Reserve University School of Medicine, Fourth Floor, Wolstein Research Building, 2103 Cornell Rd., Cleveland, OH 44106-7286. Phone: (216) 368-0508. Fax: (216) 368-4825. E-mail: paz{at}po.cwru.edu.

Journal of Clinical Microbiology, June 2004, p. 2403-2410, Vol. 42, No. 6
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.6.2403-2410.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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