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Development, validation and evaluation of a rapid PCR-nucleic acid lateral flow immuno-assay for the detection of Plasmodium and the differentiation between Plasmodium falciparum and Plasmodium vivax

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dc.contributor.author Mens, P. F.
dc.contributor.author Moers, A.
dc.contributor.author de Bes, L. M.
dc.contributor.author Flint, J.
dc.contributor.author Sak, J .R.
dc.contributor.author Keereecharoen, L.
dc.contributor.author Van Overmeir, C.
dc.contributor.author Verweij, J. J.
dc.contributor.author Hallett, R. L.
dc.contributor.author Wihokhoen, B.
dc.contributor.author Proux, S.
dc.contributor.author Schallig, H. D.
dc.contributor.author van Amerongen, A.
dc.date.accessioned 2012-12-12T13:12:54Z
dc.date.available 2012-12-12T13:12:54Z
dc.date.issued 2012
dc.identifier.issn 1475-2875
dc.identifier.doi http://dx.doi.org/10.1186/1475-2875-11-279
dc.identifier.other ITG-B7B
dc.identifier.other DBM
dc.identifier.other U-MALAR
dc.identifier.other JIF
dc.identifier.other DOI
dc.identifier.other FTA
dc.identifier.other E-only
dc.identifier.other URL
dc.identifier.other Abstract
dc.identifier.other UPD53
dc.identifier.uri http://hdl.handle.net/10390/7242
dc.description.abstract BACKGROUND: Molecular tools are very sensitive and specific and could be an alternative for the diagnosis of malaria. The complexity and need for expensive equipment may hamper implementation and, therefore, simplifications to current protocols are warranted. METHODS: A PCR detecting the different Plasmodium species and differentiating between Plasmodium falciparum and Plasmodium vivax was developed and combined with a nucleic acid lateral flow immuno-assay (PCR-NALFIA) for amplicon detection. The assay was thoroughly evaluated for the analytical sensitivity and specificity in the laboratory, the robustness and reproducibility in a ring trial and accuracy and predictive value in a field trial. RESULTS: The analytical sensitivity and specificity were 0.978 (95% CI: 0.932-0.994) and 0.980 (95% CI: 0.924-0.997), respectively, and were slightly less sensitive for the detection of P. vivax than for P. falciparum. The reproducibility tested in three laboratories was very good (k = 0.83). This evaluation showed that the PCR machine used could influence the results. Accuracy was evaluated in Thailand and compared to expert microscopy and rapid diagnostic tests (RDTs). The overall and P. falciparum-specific sensitivity and specificity was good ranging from 0.86-1 and 0.95-0.98 respectively, compared to microscopy. Plasmodium vivax detection was better than the sensitivity of RDT, but slightly less than microscopy performed in this study. CONCLUSION: PCR-NALFIA is a sensitive, specific and robust assay able to identify Plasmodium species with good accuracy. Extensive testing including a ring trial can identify possible bottlenecks before implementation and is therefore essential to perform in additon to other evaluations. en
dc.language English en
dc.subject Protozoal diseases en
dc.subject Malaria en
dc.subject Plasmodium falciparum en
dc.subject Plasmodium vivax en
dc.subject Vectors en
dc.subject Mosquitoes en
dc.subject Anopheles en
dc.subject Detection en
dc.subject Differentiation en
dc.subject Polymerase chain reaction en
dc.subject PCR en
dc.subject Immunoassay en
dc.subject Nucleic acids en
dc.subject Lateral flow tests en
dc.subject Development en
dc.subject Validation en
dc.subject Evaluation en
dc.subject Sensitivity en
dc.subject Specificity en
dc.subject Laboratory techniques and procedures en
dc.title Development, validation and evaluation of a rapid PCR-nucleic acid lateral flow immuno-assay for the detection of Plasmodium and the differentiation between Plasmodium falciparum and Plasmodium vivax en
dc.type Article-E en
dc.citation.issue 279 en
dc.citation.jtitle Malaria Journal en
dc.citation.volume 11 en
dc.citation.pages 1-10 en
dc.identifier.pmid http://www.ncbi.nlm.nih.gov/pubmed/22900750
dc.identifier.url http://www.malariajournal.com/content/11/1/279
dc.citation.jabbreviation Malar J en


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