Table 3.
Confirmation of the LOD using pooled clinical samples. Known positive samples with a Ct values ranging from 27 to 31 were used to generate pools of 8, 16, 24, 36 or 48 using 200 µL of known positive sample combined with negative saliva matrix. A total of 100 µL or 200 µL is used for RNA extraction for individual samples and pooled samples, respectively.
| Mean Ct values (SD; N = 3) |
|||||||
|---|---|---|---|---|---|---|---|
| Sample No. | Pool Size | N | MS2 | N gene | ORF1ab | S gene | % Triggering reflex |
| 1 | 1 | 3 | 28.02(0.16) | 27.50(0.73) | 29.57(0.91) | 28.96(0.76) | N/A |
| 8 | 4 | 28.05(0.19) | 28.46(0.16) | 30.69(0.34) | 28.54(0.56) | 100% | |
| 16 | 4 | 28.93(0.58) | 29.97(0.59) | 31.62(0.72) | 28.71(0.59) | 100% | |
| 24 | 12 | 28.45(0.69) | 29.29(1.04) | 31.72(0.41) | 28.9(1.33) | 100% | |
| 32 | 4 | 28.44(0.47) | 29.7(0.71) | 31.55(0.25) | 30.02(0.32) | 100% | |
| 48 | 4 | 28.52(0.5) | 30.04(1.02) | 32.08(0.84) | 29.17(0.81) | 100% | |
| 2 | 1 | 3 | 26.03(0.41) | 26.09(0.21) | 28.32(0.29) | 28.34(2.39) | N/A |
| 8 | 3 | 28.39(0.24) | 27.74(1.74) | 30.25(1.42) | 28.25(1.58) | 100% | |
| 16 | 3 | 25.96(0.97) | 28.64(0.75) | 31.43(0.08) | 29.63(0.69) | 100% | |
| 24 | 12 | 27.67(0.63) | 29.03(0.97) | 32.19(0.4) | 28.48(1.35) | 100% | |
| 32 | 3 | 28.28(0.8) | 29.6(0.39) | 33.13(1) | 29.2(0.09) | 100% | |
| 48 | 3 | 26.96(0.67) | 30.03(0.8) | 33.45(0.2) | 30.13(0.64) | 100% | |
| 3 | 1 | 3 | 27.93(0.06) | 31.13(0.58) | 28.92(1.62) | 27.95(2.31) | N/A |
| 8 | 3 | 29.05(0.22) | 29(0.52) | 30.53(0.3) | 28.89(0.16) | 100% | |
| 24 | 9 | 28.82(1.29) | 30.16(0.85) | 31.9(0.61) | 30.25(0.88) | 100% | |
| 32 | 3 | 31.06(0.4) | 31.49(1.34) | 33.74(0.15) | 32.58(0.21) | 100% | |
| 48 | 3 | 29.61(0.22) | 31.03(0.89) | 33.85(0.54) | 31.02(1.35) | 100% | |
| 4 | 1 | 3 | 25.13(0.03) | 26.89(0.15) | 29.35(0.15) | 27.71(0.24) | N/A |
| 8 | 3 | 26.93(0.06) | 28.84(0.15) | 30.81(0.07) | 29.2(0.18) | 100% | |
| 16 | 3 | 25.19(0.02) | 28.41(0.08) | 30.93(0.07) | 29.48(0.45) | 100% | |
| 24 | 3 | 24.77(0.06) | 29.1(0.17) | 31.17(0.26) | 29.56(0.14) | 100% | |
| 5 | 1 | 3 | 25.88(0.23) | 28.39(0.1) | 31.25(0.21) | 29.75(0.21) | N/A |
| 8 | 3 | 27.13(0.13) | 29.35(0.39) | 33.19(0.39) | 31.46(0.59) | 100% | |
| 16 | 3 | 25.72(0.06) | 29.9(0.11) | 32.53(0.32) | 31.1(0.32) | 100% | |
| 24 | 3 | 26.87(0.07) | 30.11(0.46) | 33.16(0.6) | 31.55(1.22) | 100% | |
| 6 | 1 | 3 | 25.86(1) | 31.54(0.58) | 31.15(0.14) | 31.62(1.03) | N/A |
| 8 | 3 | 27.91(0.11) | 31.61(0.67) | 33.36(0.18) | 32.45(0.33) | 100% | |
| 24 | 9 | 28.55(1.35) | 31.9(0.76) | 33.91(0.81) | 32.26(1) | 100% | |
| 7 | 1 | 3 | 26.65(0.9) | 26.32(0.24) | 26.44(0.04) | 26.36(0.33) | N/A |
| 24 | 6 | 25.41(0.42) | 28.09(0.32) | 29.63(0.19) | 27.76(0.28) | 100% | |
| 8 | 1 | 3 | 28.97(0.1) | 29.77(0.01) | 29.05(0.29) | 28.59(0.63) | N/A |
| 24 | 6 | 28.85(1.18) | 31.96(0.29) | 32.85(0.35) | 29.42(1.14) | 100% | |
| 9 | 1 | 3 | 31.05(1.46) | 30.25(1.18) | 29.55(0.44) | 30.33(0.62) | N/A |
| 24 | 6 | 27.59(0.54) | 31.11(0.34) | 31.5(0.2) | 29.47(0.14) | 100% | |