Table 1.
Limit of detection (LOD) confirmation using 625 and 313 viral genome copy equivalents per ml (GCE/ml) of individual specimens. A total of twenty independent RNA extractions were performed, followed by RT-PCR reactions, each performed in triplicate for each RNA extraction.
| Viral Conc | Replicate | Mean Ct (SD; N = 3) |
Interpretation | % Pos | ||||
|---|---|---|---|---|---|---|---|---|
| ORF1ab | N | S | MS2 | RP | ||||
| 625 GCE/ml (3.1 GCE/rxn) |
1 | 28.21(1.8) | 25.66(NA) | ND | 28.7(0.68) | 25.58(0.05) | Positive | 100% |
| 2 | 31.09(0.15) | ND | 29.79(NA) | 29.44(1.23) | 25.28(0.08) | Positive | ||
| 3 | 30.69(0.59) | 30.86(NA) | 28(NA) | 29.9(0.7) | 25.41(0.09) | Positive | ||
| 4 | 31.26(0.95) | ND | 29.82(NA) | 29.74(0.65) | 25.43(0.22) | Positive | ||
| 5 | 30.75(0.97) | ND | 29.85(0.64) | 29.99(0.84) | 25.6(0.06) | Positive | ||
| 6 | 30.56(0.32) | 30.12(NA) | 29.99(0.19) | 28.95(0.31) | 25.13(0.65) | Positive | ||
| 7 | 31.75(1.04) | ND | 29.31(2.36) | 29.16(0.76) | 25.68(0.13) | Positive | ||
| 8 | 30.07(1.41) | 31.46(0.81) | ND | 28.64(0.44) | 25.67(0.29) | Positive | ||
| 9 | 31.38(0.57) | 28.72(0.96) | 29.89(0.05) | 28.89(0.72) | 25.33(0.04) | Positive | ||
| 10 | 31.12(0.26) | 31.38(NA) | 29.53(NA) | 29.44(0.68) | 25.42(0.12) | Positive | ||
| 11 | 31.01(0.85) | ND | 30.7(0.25) | 29.82(0.69) | 25.26(0.13) | Positive | ||
| 12 | 30.66(0.92) | ND | 29.58(NA) | 29.71(0.55) | 25.23(0.1) | Positive | ||
| 13 | 30.05(1.04) | ND | 28.83(NA) | 30.03(0.58) | 25.45(0.23) | Positive | ||
| 14 | 30.78(0.01) | ND | 30.43(NA) | 29.87(0.66) | 25.85(0.06) | Positive | ||
| 15 | 29.07(3.14) | ND | 29.4(NA) | 29.1(0.71) | 25.55(0.27) | Positive | ||
| 16 | 30.6(0.07) | 31.15(NA) | 28.29(0.47) | 28.88(0.69) | 25.62(0.08) | Positive | ||
| 17 | 30.83(NA) | ND | 30.26(NA) | 28.76(0.56) | 25.43(0.09) | Positive | ||
| 18 | 31.2(0.81) | 30.74(NA) | ND | 29.38(0.72) | 25.62(0.07) | Positive | ||
| 19 | 32.07(0.7) | ND | 30.91(NA) | 29.58(0.78) | 25.61(0.08) | Positive | ||
| 20 | 31.75(0.79) | 32.07(NA) | ND | 29.35(0.88) | 25.17(0.22) | Positive | ||
| 313 GCE/ml (1.6 GCE/rxn |
1 | 30.22(0.94) | 30.06(NA) | 30.79(0.53) | 29.72(0.58) | 25.37(0.19) | Positive | 55% |
| 2 | 31.15(0.6) | 28.85(0.93) | ND | 29.63(0.83) | 25.49(0.1) | Positive | ||
| 3 | 31.22(0.05) | 28.9(NA) | ND | 29.26(0.76) | 25.54(0.1) | Positive | ||
| 4 | 30.06(0.69) | 34.79(NA) | ND | 28.29(0.6) | 25.07(0.41) | Positive | ||
| 5 | 30.49(NA) | ND | 30.79(NA) | 28.7(0.59) | 25.32(0.05) | Positive | ||
| 6 | ND | ND | ND | 29.35(0.77) | 25.46(0.1) | Negative | ||
| 7 | 31.55(1.28) | ND | ND | 29.4(0.91) | 25.36(0.07) | Inconclusive | ||
| 8 | 30.45(0.33) | ND | 29.7(NA) | 29.14(0.82) | 25.02(0.11) | Positive | ||
| 9 | 31.64(0.78) | ND | 31.66(NA) | 29.43(1.04) | 25.37(0.05) | Positive | ||
| 10 | 30.55(0.88) | ND | 30.55(NA) | 29.38(0.68) | 25.59(0.13) | Positive | ||
| 11 | 31.07(0.83) | ND | ND | 28.94(0.87) | 25.54(0.19) | Inconclusive | ||
| 12 | 31.28(1.69) | ND | ND | 28.48(0.92) | 25.66(0.07) | Inconclusive | ||
| 13 | 31.14(1.36) | ND | ND | 28.34(0.82) | 25.21(0.07) | Inconclusive | ||
| 14 | 29.81(0.14) | 30.19(NA) | 27.77(NA) | 29.23(0.94) | 25.38(0.05) | Positive | ||
| 15 | 30.99(1.42) | ND | 30.17(NA) | 29.41(0.97) | 25.14(0.06) | Positive | ||
| 16 | 32.29(0.39) | ND | ND | 29.39(1.04) | 25.27(0.13) | Inconclusive | ||
| 17 | 29.59(0.33) | 30.58(0.65) | 29.85(NA) | 29.25(0.89) | 25.3(0.1) | Positive | ||
| 18 | 30.56(1.45) | ND | 28.59(NA) | 28.98(0.79) | 25.42(0.08) | Positive | ||
| 19 | 29.94(0.28) | ND | ND | 29.12(0.9) | 25.53(0.12) | Inconclusive | ||
| 20 | 30.51(0.34) | ND | ND | 28.36(0.87) | 25.33(0.03) | Inconclusive | ||
N = number of RNA extractions; RP = Hs_RPP30 primer for targeting human RNase P.