The online version of this article (https://doi.org/10.1186/s12936-018-2201-0) contains supplementary material, which is available to authorized users.
Maria Gruenberg and Clara Antunes Moniz contributed equally to this work
A distinctive feature of Plasmodium vivax infections is the overall low parasite density in peripheral blood. Thus, identifying asymptomatic infected individuals in endemic communities requires diagnostic tests with high sensitivity. The detection limits of molecular diagnostic tests are primarily defined by the volume of blood analysed and by the copy number of the amplified molecular marker serving as the template for amplification. By using mitochondrial DNA as the multi-copy template, the detection limit can be improved more than tenfold, compared to standard 18S rRNA targets, thereby allowing detection of lower parasite densities. In a very low transmission area in Brazil, application of a mitochondrial DNA-based assay increased prevalence from 4.9 to 6.5%. The usefulness of molecular tests in malaria epidemiological studies is widely recognized, especially when precise prevalence rates are desired. Of concern, however, is the challenge of demonstrating test accuracy and quality control for samples with very low parasite densities. In this case, chance effects in template distribution around the detection limit constrain reproducibility. Rigorous assessment of false positive and false negative test results is, therefore, required to prevent over- or under-estimation of parasite prevalence in epidemiological studies or when monitoring interventions.
Additional file 1: Table S1. Gene IDs of P. vivax 18S rRNA (small subunit rRNA gene).
Additional file 2: Table S2. Assay conditions for P. vivax qPCR targeting cox1 (Pv-mtCOX1 assay).
Additional file 3: Table S3. Performance Pv-mtCOX1 qPCR.
Additional file 4: Table S4. Limit of detection of Pv-mtCOX1.
Additional file 5: Figure S1. Fold-difference in template copy numbers detected by molecular marker Pv-mtCOX1 versus marker Pv18S rRNA.
Additional file 6: Figure S2. LAMP reaction detected with hydroxynaphtol blue (HNB).
Additional file 7: Figure S3. Real-time LAMP reaction with calcein detection.
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- Plasmodium vivax molecular diagnostics in community surveys: pitfalls and solutions
Clara Antunes Moniz
Natalie Ellen Hofmann
Wuelton Marcelo Monteiro
Gisely Cardoso de Melo
Andre M. Siqueira
- BioMed Central
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