Table 3 The efficiency and analytical sensitivity for the Babesia LSU qPCR. The efficiency and analytical sensitivity were determined for the Babesia LSU qPCR using plasmids as template DNA run in 20 intra-assay replicates at 3 copies/well and 5 copies/well. Assays were run with all 3 primers (Bmic-F, B-lsu-F and B-lsu-R2) for both plasmids and each corresponding 2 primer reaction: Bmic-F with B-lsu-R2 for B. microti-like plasmid (pBMIC-LSU) and B-lsu-F with B-lsu-R2 for B. vogeli plasmid (pBCV-LSU)
Primers | Plasmid template | Eff (%) | R 2 | 3 c/well (%) | Cq (range) | 5 c/well (%) | Cq (range) |
|---|---|---|---|---|---|---|---|
2 primers | pBMIC-LSU | 100 | 0.99 | 90 | 34.1–37.2 | 100 | 33.5–37.5 |
3 primers | pBMIC-LSU | 95 | 0.99 | 70 | 33.6–37.3 | 95 | 34.2–37.8 |
2 primers | pBCV-LSU | 94 | 0.99 | 60 | 37.1–39.9 | 100 | 33.3–36.7 |
3 primers | pBCV-LSU | 92 | 0.99 | 60 | 36.8–39.2 | 100 | 33–34.2 |