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. 2019 Sep 11;9:13107. doi: 10.1038/s41598-019-49389-2

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Multiplex detection of pfhrp2 and pfhrp3 genes using the qHRP2/3-del assay. (A) The qHRP2/3-del assay amplifies pfhrp2, pfhrp3 and pfrnr2e2 target sequences in a multiplex qPCR reaction and correctly identifies strains carrying either a pfhrp2 deletion (PfDD2), a pfhrp3 deletion (PfHB3) or no deletion (PfNF54). (B) Performance characteristic of each individual amplification assay, run within the multiplex qHRP2/3-del assay, is shown. Correlation with high linearity between serially diluted WHO international standard for P. falciparum NATs (PfIS) and Cq values was obtained and used to calculate the qPCR efficiency. Cq values above 40 (black line) are considered negative.