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. 2021 Aug 5;11:15962. doi: 10.1038/s41598-021-95402-y

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© The Author(s) 2021

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Specificity assay among Anaplasma species electrophoresed products obtained by RPA-basic primers in the top panel and the multiplex RPA-nfo reactions in corresponding PCRD lateral flow devices in the bottom panel. (A) Gel-based RPA primers. Lane L, 100 bp DNA ladder; lane Am, A. marginale; lane Ao, A. ovis; lane Ap, A. phagocytophilum; lane T, lab-reared tick DNA; lane N, non-template control (NTC, water). (B) Multiplex RPA-nfo primers and probes. Lane C, flow-check line; Lane 2, detects FAM/Biotin labelled amplicons (Anaplasma species); Lane 1, detects DIG/Biotin labelled amplicons (GAPDH—internal control); Am, A. marginale; Ao, A. ovis; Ap, A. phagocytophilum; T, lab-reared tick DNA; N, non-template control (water).