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01.12.2012 | Research | Ausgabe 1/2012 Open Access

Malaria Journal 1/2012

Real-time PCR assay for discrimination of Plasmodium ovale curtisi and Plasmodium ovale wallikeri in the Ivory Coast and in the Comoros Islands

Zeitschrift:
Malaria Journal > Ausgabe 1/2012
Autoren:
Frédérique Bauffe, Jérôme Desplans, Christophe Fraisier, Daniel Parzy
Wichtige Hinweise

Electronic supplementary material

The online version of this article (doi:10.​1186/​1475-2875-11-307) contains supplementary material, which is available to authorized users.
Frédérique Bauffe, Jérôme Desplans contributed equally to this work.

Competing interests

The authors declare that they have no competing interests.

Authors’ contributions

FB, CF and JD performed the sequencing and carried out the molecular genetic studies. FB and JD performed the RDT and designed the probes. DP and JD conceived and funded the project. FB and JD prepared the first draft of the paper and all authors contributed to the writing of the report and have reviewed and approved the final version.

Abstract

Background

Plasmodium ovale is one of the five malaria species infecting humans. Recent data have shown that the name of this neglected species masks two distinct genotypes also called curtisi and wallikeri. Some authors show that these species could be sympatric. These two subspecies are not differentiated by microscopy techniques and malaria rapid diagnostic tests. This diagnostic defect is the result of low parasitaemia, antigenic polymorphism and absence of antibodies performance and requires the use of sequencing techniques. An accurate and easy discrimination detection method is necessary.

Methods

A new molecular assay was developed to easily identify the two genotypes of P. ovale. This tool allowed the study of 90 blood samples containing P. ovale, confirmed by molecular biology techniques, which were obtained from patients with imported malaria.

Results

The new marker was validated on well genotyped samples. The genotype of 90 P. ovale samples mainly imported from the Ivory Coast and the Comoros Islands was easily and quickly realized. The distribution of the two subspecies was described with a significant number of samples and showed that the two genotypes were present in the studied countries.

Conclusion

This work confirms the presence of the two species in the same country for the first time, in the Ivory Coast and the Comoros Islands. A better genotyping of P. ovale types may improve a better characterization of the clinical pathophysiology for each.
Zusatzmaterial
Additional file 1: Alignment of PocLDH and PowLDH. Using PoLDH (Genbank AY486058) as reference, obtained sequences for P. o. curtisi (Poc_LDH) and P. o. wallikeri (Pow_LDH) were aligned in nucleotide (A) and translated protein (B) with CLUSTAL 2.0.12 multiple sequence alignment software. The Sequence PocLDH and PowLDH were obtained from samples Po22 and Po23 respectively. In A, the reference sequence AY486058 is related to P. o. curtisi. Differences in Pow sequences are box-shaded in grey. In (B), the full sequence of Pf-pLDH protein was added like another reference (Pf13_141). The amino-acid switch between Poc and Pow sequences are indicated with their amino-acid number. All other differences are box-shaded, conservative in grey and non-conservative in white. (PDF 46 KB)
12936_2012_2465_MOESM1_ESM.pdf
Additional file 2: Detail of the sequencing results per sample. The genotyping result is indicated for each sample (P. o. curtisi (C) and P. o. wallikeri (W)) with reference sequences obtained by ldh or SSuRNA and results of POCPOW marker. Corresponding parasitaemia and RDT results according to the antigen used are indicated. neg: RDT negative result; pos: RDT positive result, T2: pan signal. (DOC 126 KB)
12936_2012_2465_MOESM2_ESM.doc
Authors’ original file for figure 1
12936_2012_2465_MOESM3_ESM.pdf
Literatur
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