Evaluation of a novel real-time PCR assay for the detection, identification and quantification of Plasmodium species causing malaria in humans
Abstract Background The entry of PCR-based techniques into malaria diagnostics has improved the sensitivity and specificity of the detection of Plasmodium infections. It has been shown that humans are regularly infected by at least six different Plasmodium species. The MC004 real-time PCR assay for...
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ftdoajarticles:oai:doaj.org/article:f95ace76bced4b4f9b629c795136b9c8 2023-05-15T15:14:12+02:00 Evaluation of a novel real-time PCR assay for the detection, identification and quantification of Plasmodium species causing malaria in humans Kim van Bergen Toon Stuitje Robert Akkers Eric Vermeer Rob Castel Theo Mank 2021-07-01T00:00:00Z https://doi.org/10.1186/s12936-021-03842-8 https://doaj.org/article/f95ace76bced4b4f9b629c795136b9c8 EN eng BMC https://doi.org/10.1186/s12936-021-03842-8 https://doaj.org/toc/1475-2875 doi:10.1186/s12936-021-03842-8 1475-2875 https://doaj.org/article/f95ace76bced4b4f9b629c795136b9c8 Malaria Journal, Vol 20, Iss 1, Pp 1-12 (2021) Malaria Real-time PCR Melting curve analysis Identification and quantification of Plasmodium species Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 article 2021 ftdoajarticles https://doi.org/10.1186/s12936-021-03842-8 2022-12-31T04:29:13Z Abstract Background The entry of PCR-based techniques into malaria diagnostics has improved the sensitivity and specificity of the detection of Plasmodium infections. It has been shown that humans are regularly infected by at least six different Plasmodium species. The MC004 real-time PCR assay for malaria diagnosis is a novel single-tube assay that has been developed for the purpose of simultaneously detecting all Plasmodium species known to infect humans, and discrimination between Plasmodium falciparum, Plasmodium vivax, Plasmodium malariae, Plasmodium ovale wallikeri, Plasmodium ovale curtisi, Plasmodium knowlesi (including differentiation of three strains) and Plasmodium cynomolgi (including differentiation of three strains). Detection and identification of Plasmodium species relies on molecular beacon probe-based melting curve analysis. In addition, this assay might be used to quantify the parasitaemia of at least P. falciparum by calculating the level of parasitaemia directly from the Cq-value. Methods The samples used in this study comprised reference samples, patient samples, and synthetic controls. The following analytical performance characteristics of the MC004 assay were determined: analytical specificity, limit of detection, the ability to detect mixed infections, and the potential to determine the level of parasitaemia of P. falciparum, including assessment of within-run and between-run precisions. Results No false positive or false negative results were observed. The limit of detection of P. falciparum was 1 × 10–3 IU/mL (WHO standard). Mixed infections with P. falciparum and non-falciparum species were correctly identified. A calibration curve could be established to quantify the parasitaemia of at least P. falciparum. The within-run and between-run precisions were less than 20% CV at the tested parasitaemia levels of 0.09%, 0.16%, 2.15% and 27.27%. Conclusion Based upon the analytical performance characteristics that were determined, the MC004 assay showed performance suitable for use in ... Article in Journal/Newspaper Arctic Directory of Open Access Journals: DOAJ Articles Arctic Malaria Journal 20 1 |
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Directory of Open Access Journals: DOAJ Articles |
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ftdoajarticles |
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English |
topic |
Malaria Real-time PCR Melting curve analysis Identification and quantification of Plasmodium species Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 |
spellingShingle |
Malaria Real-time PCR Melting curve analysis Identification and quantification of Plasmodium species Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 Kim van Bergen Toon Stuitje Robert Akkers Eric Vermeer Rob Castel Theo Mank Evaluation of a novel real-time PCR assay for the detection, identification and quantification of Plasmodium species causing malaria in humans |
topic_facet |
Malaria Real-time PCR Melting curve analysis Identification and quantification of Plasmodium species Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 |
description |
Abstract Background The entry of PCR-based techniques into malaria diagnostics has improved the sensitivity and specificity of the detection of Plasmodium infections. It has been shown that humans are regularly infected by at least six different Plasmodium species. The MC004 real-time PCR assay for malaria diagnosis is a novel single-tube assay that has been developed for the purpose of simultaneously detecting all Plasmodium species known to infect humans, and discrimination between Plasmodium falciparum, Plasmodium vivax, Plasmodium malariae, Plasmodium ovale wallikeri, Plasmodium ovale curtisi, Plasmodium knowlesi (including differentiation of three strains) and Plasmodium cynomolgi (including differentiation of three strains). Detection and identification of Plasmodium species relies on molecular beacon probe-based melting curve analysis. In addition, this assay might be used to quantify the parasitaemia of at least P. falciparum by calculating the level of parasitaemia directly from the Cq-value. Methods The samples used in this study comprised reference samples, patient samples, and synthetic controls. The following analytical performance characteristics of the MC004 assay were determined: analytical specificity, limit of detection, the ability to detect mixed infections, and the potential to determine the level of parasitaemia of P. falciparum, including assessment of within-run and between-run precisions. Results No false positive or false negative results were observed. The limit of detection of P. falciparum was 1 × 10–3 IU/mL (WHO standard). Mixed infections with P. falciparum and non-falciparum species were correctly identified. A calibration curve could be established to quantify the parasitaemia of at least P. falciparum. The within-run and between-run precisions were less than 20% CV at the tested parasitaemia levels of 0.09%, 0.16%, 2.15% and 27.27%. Conclusion Based upon the analytical performance characteristics that were determined, the MC004 assay showed performance suitable for use in ... |
format |
Article in Journal/Newspaper |
author |
Kim van Bergen Toon Stuitje Robert Akkers Eric Vermeer Rob Castel Theo Mank |
author_facet |
Kim van Bergen Toon Stuitje Robert Akkers Eric Vermeer Rob Castel Theo Mank |
author_sort |
Kim van Bergen |
title |
Evaluation of a novel real-time PCR assay for the detection, identification and quantification of Plasmodium species causing malaria in humans |
title_short |
Evaluation of a novel real-time PCR assay for the detection, identification and quantification of Plasmodium species causing malaria in humans |
title_full |
Evaluation of a novel real-time PCR assay for the detection, identification and quantification of Plasmodium species causing malaria in humans |
title_fullStr |
Evaluation of a novel real-time PCR assay for the detection, identification and quantification of Plasmodium species causing malaria in humans |
title_full_unstemmed |
Evaluation of a novel real-time PCR assay for the detection, identification and quantification of Plasmodium species causing malaria in humans |
title_sort |
evaluation of a novel real-time pcr assay for the detection, identification and quantification of plasmodium species causing malaria in humans |
publisher |
BMC |
publishDate |
2021 |
url |
https://doi.org/10.1186/s12936-021-03842-8 https://doaj.org/article/f95ace76bced4b4f9b629c795136b9c8 |
geographic |
Arctic |
geographic_facet |
Arctic |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
Malaria Journal, Vol 20, Iss 1, Pp 1-12 (2021) |
op_relation |
https://doi.org/10.1186/s12936-021-03842-8 https://doaj.org/toc/1475-2875 doi:10.1186/s12936-021-03842-8 1475-2875 https://doaj.org/article/f95ace76bced4b4f9b629c795136b9c8 |
op_doi |
https://doi.org/10.1186/s12936-021-03842-8 |
container_title |
Malaria Journal |
container_volume |
20 |
container_issue |
1 |
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1766344677435375616 |