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. 2024 Nov 6;14:26884. doi: 10.1038/s41598-024-77164-5

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© The Author(s) 2024

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Assay Optimization. Shown are Cq values measured from an ATB participant sample. a, Assay multiplexing: SP - Single-target reaction, DP - Double-target reaction and MP - Multiplex, mean expression levels. Comparison was performed using the Kruskal-Wallis test and Dunn’s Test with Bonferroni corrected p-values for multiple comparisons. A p-value of 0.05 or less was considered significant and * is equivalent to p < 0.05. b, Assay precision testing line graphs showing assay read-out Cq values. Assay technical repeat runs 1–6 were performed on 4th of November 2022 and assay technical repeat runs 7 to 14 were performed on 6th February 2023. Percentage values within the graph represent the Cq CVs between runs for target genes.