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Plasmodium ovale malaria has been reported in various countries in southeast Asia, but never in Cambodia. Using a species-specific polymerase chain reaction (PCR) targeting the small subunit (SSU) ribosomal RNA (rRNA) gene, we detected P. ovale in nearly 4% of the inhabitants of a northeastern Cambodian village. Plasmodium ovale was associated with at least one other Plasmodium species, and two quadruple infections were detected. The diagnosis was confirmed by microscopy and by SSU rRNA PCR product sequencing. The sequences shared 96–99% identity with published sequences, and displayed a substantial heterogeneity with 2–4 different haplotypes per sample. Nine distinct SSU rRNA haplotypes were identified, including seven novel variants. Phylogenetic analysis showed two major genetic clusters, suggesting amplification of two distinct gene sets and/or P. ovale variants from each sample. Our data indicate that P. ovale was overlooked in Cambodia until now, and call for the implementation of larger prevalence surveys and accurate diagnosis methods in this country.
Received April 7, 2004. Accepted for publication November 12, 2004.
Acknowledgments: We thank the staff of the National Center for Parasitology, Entomology and Malaria Control as well as the staff of the European Commission National Malaria Control Program for sample collection and for the initial examination of the blood slides.
Financial support: This work was supported by the PAL+ program of the French Ministry of Research and New Technologies.
Authors’ addresses: Sandra Incardona, Sophy Chy, Sina Nhem, and Rithy Sem, Institut Pasteur du Cambodge, 5 Boulevard Monivong, BP 983, Phnom Penh, Cambodia, Telephone: 855-23-368-036, Fax: 855-23-725-606, E-mails: sandra{at}pasteur-kh.org, chsophy{at}pasteur-kh.org, sandra{at}pasteur-kh.org, and srithy{at}pasteur-kh.org. Lim Chiv, Sean Hewitt, and Socheat Doung, National Center for Parasitology, Entomology and Malaria Control, 372 Boulevard Monivong/Rue 322, Phnom Penh, Cambodia, Telephone: 855-23-219-271, Fax: 855-23-219-271, E-mails: cnm{at}bigpond.com.kh, hewitt_se{at}hotmail.com, and socheatd{at}cnm.gov.kh. Odile Mercereau-Puijalon, Unité d’Immunologie Moléculaire des Parasites, Centre National de la Recherche Scientifique, Unité de Recherche Associee 2581, Institut Pasteur, 25 Rue du Docteur Roux, 75724 Paris Cedex 15, France, Telephone: 33-1-45-68-86-23, Fax: 33-1-40-61-35-41, E-mail: omp{at}pasteur.fr. Thierry Fandeur, Unité Mixte de Recherche, Institut National de Recherche Agronomique d’Immunologie Parasitaire, Faculté des Sciences Pharmaceutiques, 31 Avenue Monge, Parc Grandmont, 37200 Tours, France, E-mail: thierry.fandeur{at}univ-tours.fr.
Reprint requests: Sandra Incardona, Institut Pasteur du Cambodge, 5 Boulevard Monivong, BP 983, Phnom Penh, Cambodia.
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