Table 2.
Summary of LOD preliminary range finding and confirmation in pooled specimens using known concentrations of ATCC VR-1986HK whole inactivated virus spiked into individual negative saliva. The stock concentration provided by ATCC was 4.2 × 105 GCE/µL. For each RNA extraction, 200 µL of each sample was used.
| Viral Conc | Pool size | N | Mean Ct (SD; N = 3) |
Result | % Triggering Reflex | ||||
|---|---|---|---|---|---|---|---|---|---|
| ORF1ab | N gene | S gene | MS2 | RP | |||||
| 1250 GCE/ml (6.3 GCE/rxn) |
1 | 1 | 29.96(0.3) | 28.37(0.34) | 24.94(4.28) | 28.67(1.06) | 23.29(0.45) | Positive | 100% |
| 2 | 29.99(0.43) | 28.51(1.44) | 25.75(5.33) | 28.11(0.47) | 23.04(0.04) | Positive | |||
| 3 | 30.67(1.01) | 28.86(2) | 26.36(2.34) | 28.27(0.28) | 22.88(0.16) | Positive | |||
| 8 | 1 | 32.83(0.62) | 30.18(0.31) | 30.25(0.75) | 27.9(0.5) | 23.19(0.08) | Positive | 100% | |
| 2 | 31.97(0.64) | 30.4(0.1) | 27.73(1.87) | 28.24(0.41) | 23.32(0.11) | Positive | |||
| 3 | 32.36(1.18) | 30.11(2.73) | 25.88(4.4) | 28.51(0.4) | 23.23(0.07) | Positive | |||
| 16 | 1 | 33.37(0.6) | 31.89(1.04) | 33.27(NA) | 27.98(0.57) | 23.02(0.07) | Positive | 100% | |
| 2 | 32.28(NA) | ND | ND | 28.4(0.93) | 23.12(0.02) | Inconclusive | |||
| 3 | ND | 31.38(NA) | 28.46(NA) | 28.9(0.59) | 23.19(0.14) | Positive | |||
| 24 | 1 | 32.65(1.13) | 31.87(NA) | 31.05(NA) | 28.16(0.92) | 23(0.04) | Positive | 95.83% | |
| 2 | 33.74(0.03) | 30.8(NA) | ND | 28.54(0.86) | 23.08(0.04) | Positive | |||
| 3 | 33.76(0.43) | ND | 30.01(0.9) | 29.22(0.37) | 23.03(0.11) | Positive | |||
| 4 | 31.29(NA) | 30.5(NA) | ND | 27.23(0.97) | 22.72(0.05) | Positive | |||
| 5 | 32.58(0.12) | 32.47(NA) | ND | 27.04(0.7) | 22.79(0.22) | Positive | |||
| 6 | 32.96(0.6) | ND | 24.91(NA) | 27.34(1.11) | 22.83(0.07) | Positive | |||
| 7 | 33.76(NA) | ND | ND | 27.63(0.3) | 22.95(0.23) | Inconclusive | |||
| 8 | 30.88(NA) | ND | ND | 27.79(0.77) | 23.17(0.13) | Inconclusive | |||
| 9 | 33.62(NA) | 32.82(NA) | 32.23(0.12) | 27.83(0.37) | 22.7(0.15) | Positive | |||
| 10 | 33.52(NA) | 31.03(NA) | ND | 28.24(0.52) | 23.3(0.06) | Positive | |||
| 11 | 32.73(NA) | ND | 27.8(NA) | 27.89(0.26) | 22.98(0.12) | Positive | |||
| 12 | 33.31(1.73) | 30.85(NA) | 30.5(NA) | 28.03(0.63) | 23.1(0.03) | Positive | |||
| 13 | 32.36(NA) | 31.92(NA) | ND | 27.87(0.44) | 22.96(0.05) | Positive | |||
| 14 | 32.53(NA) | 31.87(0.29) | 30.92(NA) | 28.24(0.09) | 23.09(0.13) | Positive | |||
| 15 | 32.6(0.37) | 31.34(1.44) | 27.65(3.36) | 27.81(0.22) | 22.8(0.18) | Positive | |||
| 16 | 32.54(NA) | ND | ND | 28.37(0.57) | 23.05(0.02) | Inconclusive | |||
| 17 | ND | ND | 29.7(NA) | 27.94(0.3) | 23.14(0.1) | Inconclusive | |||
| 18 | ND | ND | ND | 28.21(0.67) | 23.05(0.03) | Negative | |||
| 19 | 33.07(NA) | ND | 30.85(1.38) | 27.91(0.21) | 23.22(0.11) | Positive | |||
| 20 | 32.86(NA) | ND | 32.46(NA) | 27.79(0.65) | 23.31(0.14) | Positive | |||
| 36 | 1 | 34.12(NA) | 29.38(NA) | ND | 28.36(0.56) | 23.02(0.05) | Positive | 100% | |
| 2 | 33.87(NA) | 30.69(2.08) | 25.13(NA) | 28.6(0.73) | 23.06(0.08) | Positive | |||
| 3 | 33.56(NA) | 25.9(0.05) | ND | 28.13(1.09) | 22.95(0.04) | Positive | |||
| 48 | 1 | 33.78(NA) | 31.32(NA) | ND | 28.05(0.76) | 23.1(0.04) | Positive | 66.67% | |
| 2 | ND | 32.12(NA) | 31.42(NA) | 28.41(0.49) | 23.35(0.05) | Positive | |||
| 3 | ND | ND | ND | 27.86(1.17) | 23.09(0.05) | Negative | |||