Table 3. Analytical sensitivity (limit of detection, LoD) and analytical precision of the multiplex qPCR assay.
| Average Ct values depending on the copy number of target DNA | ||||||||
|---|---|---|---|---|---|---|---|---|
| Target organism | Performances | 30,000 copies | 3,000 copies | 300 copies | 30 copies | 10 copies | 5 copies | N.C. |
| Cryptosporidium parvum | ||||||||
| Average Ct value | 26.05 | 29.2 | 32.9 | 36.26 | 39.31 | 39.41 | - | |
| SD | 0.11 | 0.22 | 0.45 | 0.43 | 2.22 | 1.58 | - | |
| CV | 0.41% | 0.74% | 1.37% | 1.17% | 5.65% | 4.00% | - | |
| Detection rate | 100% | 100% | 100% | 100% | 67% | 38% | - | |
| Giardia lamblia | ||||||||
| Average Ct value | 27.46 | 30.77 | 34.15 | 37.24 | 40.98 | 39.26 | - | |
| SD | 0.23 | 0.27 | 0.46 | 0.75 | 2.23 | 0.97 | - | |
| CV | 0.83% | 0.86% | 1.34% | 2.01% | 5.45% | 2.47% | - | |
| Detection rate | 100% | 100% | 100% | 100% | 100% | 33% | 0% | |
| Entamoeba histolytica | ||||||||
| Average Ct value | 26.52 | 30.02 | 33.36 | 36.94 | 39.28 | 39.01 | - | |
| SD | 0.17 | 0.22 | 0.39 | 1.69 | 1.72 | 1.77 | - | |
| CV | 0.64% | 0.74% | 1.16% | 4.58% | 4.37% | 4.55% | - | |
| Detection rate | 100% | 100% | 100% | 96% | 54% | 25% | 0% | |
| Dientamoeba fragilis | ||||||||
| Average Ct value | 27.73 | 31.15 | 34.7 | 37.53 | 40.72 | 41.44 | - | |
| SD | 0.39 | 0.32 | 0.65 | 0.73 | 2.11 | 1.95 | - | |
| CV | 1.40% | 1.04% | 1.86% | 1.96% | 5.19% | 4.71% | - | |
| Detection rate | 100% | 100% | 100% | 96% | 63% | 71% | 0% | |
| Blastocystis hominis | ||||||||
| Average Ct value | 30.47 | 34.08 | 37.29 | 40.63 | 42.25 | 40.77 | - | |
| SD | 0.41 | 0.51 | 0.59 | 0.82 | 1.55 | 1.4 | - | |
| CV | 1.35% | 1.51% | 1.58% | 2.01% | 3.68% | 3.43% | - | |
| Detection rate | 100% | 100% | 100% | 100% | 46% | 17% | 0% | |
| Clonorchis sinensis | ||||||||
| Average Ct value | 25.73 | 29.28 | 32.23 | 36.23 | 39.47 | 39.2 | - | |
| SD | 0.12 | 0.28 | 0.35 | 1.07 | 2.68 | 1.84 | - | |
| CV | 0.48% | 0.97% | 1.09% | 2.96% | 6.78% | 4.70% | - | |
| Detection rate | 100% | 100% | 100% | 100% | 79% | 50% | 0% | |
| Metagonimus yokogawai | ||||||||
| Average Ct value | 25.88 | 29.39 | 32.24 | 35.65 | 38.38 | 39.51 | - | |
| SD | 0.09 | 0.17 | 0.65 | 1.77 | 1.31 | 2.11 | - | |
| CV | 0.35% | 0.57% | 2.02% | 4.96% | 3.42% | 5.34% | - | |
| Detection rate | 100% | 100% | 100% | 100% | 100% | 58% | 0% | |
| Gymnophalloides seoi | ||||||||
| Average Ct value | 30.15 | 33.94 | 37.2 | 40.14 | 41.64 | 40.21 | - | |
| SD | 0.45 | 0.33 | 0.42 | 0.76 | 1.56 | 1.66 | - | |
| CV | 1.51% | 0.97% | 1.12% | 1.89% | 3.74% | 4.12% | - | |
| Detection rate | 100% | 100% | 100% | 100% | 54% | 13% | 0% | |
Ct = threshold cycle; CV = coefficient of variation; N.C. = negative control; SD = standard deviation.