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Parasitol Int. 2015 Jun;64(3):304-8. doi: 10.1016/j.parint.2014.09.006. Epub 2014 Sep 22.

Improved detection of malaria cases in island settings of Vanuatu and Kenya by PCR that targets the Plasmodium mitochondrial cytochrome c oxidase III (cox3) gene.

Author information

1
Department of Medical Zoology, Graduate School of Medicine, Osaka City University, 1-4-3, Asahi-machi, Abeno-ku, Osaka 545-8585, Japan.
2
Department of Medical Zoology, Graduate School of Medicine, Osaka City University, 1-4-3, Asahi-machi, Abeno-ku, Osaka 545-8585, Japan; Island Malaria Group, Department of Microbiology, Tumor and Cell Biology (MTC), Karolinska Institutet, Nobels väg 16, SE 171 77 Stockholm, Sweden.
3
Island Malaria Group, Department of Microbiology, Tumor and Cell Biology (MTC), Karolinska Institutet, Nobels väg 16, SE 171 77 Stockholm, Sweden.
4
Division of International Health, Research Promotion Project, Oita University Faculty of Medicine, Yufu 879-5593, Japan.
5
Radioisotope Centre, Graduate School of Medicine, Osaka City University, 1-4-3, Asahi-machi, Abeno-ku, Osaka 545-8585, Japan. Electronic address: mkimura@med.osaka-cu.ac.jp.

Abstract

Detection of sub-microscopic parasitemia is crucial for all malaria elimination programs. PCR-based methods have proven to be sensitive, but two rounds of amplification (nested PCR) are often needed to detect the presence of Plasmodium DNA. To simplify the detection process, we designed a nested PCR method whereby only the primary PCR is required for the detection of the four major human Plasmodium species. Primers designed for the detection of the fifth species, Plasmodium knowlesi, were not included in this study due to the absence of appropriate field samples. Compared to the standard 18S rDNA PCR method, our cytochrome c oxidase III (cox3) method detected 10-50% more cases while maintaining high sensitivities (1.00) for all four Plasmodium species in our samples from Vanuatu (n=77) and Kenya (n=76). Improvement in detection efficiency was more substantial for samples with sub-microscopic parasitemia (54%) than those with observable parasitemia (10-16%). Our method will contribute to improved malaria surveillance in low endemicity settings.

KEYWORDS:

Human malaria; Malaria diagnosis; Mitochondrial DNA; Nested PCR; Plasmodium; cox3

PMID:
25256904
DOI:
10.1016/j.parint.2014.09.006
[Indexed for MEDLINE]

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