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. 2021 Nov 25;9(1):1–20. doi: 10.15698/mic2022.01.767

Table 3.

Sensitivities of SARS-CoV-2-specific primers and probes in fourplex rRT-PCR assays.

Fourplex 1
Probes in the fourplex E Sarbeco-FAM Orf1ab L LNA-TEX N LSPQ-HEX B2M-Cy5
Lowest numbers of copies/reaction Test 1 (Ct) Test 2 (Ct) Test 3 (Ct) Test 1 (Ct) Test 2 (Ct) Test 3 (Ct) Test 1 (Ct) Test 2 (Ct) Test 3 (Ct) Test 1 (Ct) Test 2 (Ct) Test 3 (Ct)
1.4 35.25 35.78 35.47 32.95 32.25 32.41 nd nd nd 33.97 34.05 33.85
14 33.53 33.70 33.61 29.48 29.71 29.56 37.03 36.39 36.68 30.12 30.31 30.04
140 30.97 30.70 30.52 27.33 27.81 27.52 35.55 33.69 34.11 26.64 27.01 26.55
1.4E+03 27.52 27.34 27.42 24.86 24.92 24.79 30.90 30.66 30.77 23.48 23.50 23.31
1.4E+04 23.93 23.97 23.88 21.89 21.75 21.78 27.28 27.30 27.42 19.85 19.97 19.71
1.4E+05 20.61 20.68 20.59 18.35 18.26 18.21 23.81 23.82 23.65 16.67 16.89 16.61
1.4E+06 17.17 17.11 17.08 14.91 14.86 14.77 20.27 20.24 20.14 12.55 12.76 12.52
1.4E+07 13.62 13.66 13.55 11.30 11.21 11.17 16.87 16.89 16.73 9.52 9.63 9.46
1.4E+08 10.09 10.01 9.94 7.70 7.58 7.63 13.69 13.58 13.51 6.07 6.10 6.01
1.4E+09 6.86 6.70 6.66 4.12 3.79 3.86 10.17 10.21 10.07 3.19 3.14 3.08
NTC Neg Neg Neg Neg Neg Neg Neg Neg Neg Neg Neg Neg
Fourplex 2
Probes in the fourplex E Sarbeco-FAM Orf1ab L LNA-TEX Spike 614G LNA-HEX B2M-Cy5
Lowest numbers of copies/reaction Test 1 (Ct) Test 2 (Ct) Test 3 (Ct) Test 1 (Ct) Test 2 (Ct) Test 3 (Ct) Test 1 (Ct) Test 2 (Ct) Test 3 (Ct) Test 1 (Ct) Test 2 (Ct) Test 3 (Ct)
1.4 35.58 35.46 35.51 32.72 32.61 32.66 34.92 34.75 34.81 34.44 34.36 34.31
14 32.67 32.85 32.72 30.20 30.32 30.24 32.57 32.83 32.66 31.68 31.40 31.48
140 29.67 29.55 29.61 27.49 27.42 27.36 29.56 29.77 29.61 29.39 29.43 29.36
1.4E+03 25.83 25.92 25.72 25.86 25.84 25.72 26.47 26.8 26.62 26.91 26.82 26.78
1.4E+04 22.53 22.70 22.61 22.84 23.05 22.75 21.85 22.20 21.94 23.60 23.61 23.52
1.4E+05 18.10 18.90 18.44 19.71 20.10 19.83 18.40 18.22 18.03 19.68 18.52 18.43
1.4E+06 14.96 14.93 14.84 15.32 15.60 15.28 14.55 14.60 14.42 15.32 15.45 15.25
1.4E+07 11.25 11.26 11.13 11.92 11.80 11.85 10.72 10.62 10.58 11.64 11.62 11.51
1.4E+08 7.99 7.54 7.41 8.23 8.20 8.16 7.74 7.80 7.69 7.90 7.91 7.86
1.4E+09 5.10 5.25 5.21 5.54 5.56 5.45 4.20 4.01 4.07 4.95 4.92 4.87
NTC Neg Neg Neg Neg Neg Neg Neg Neg Neg Neg Neg Neg
Fourplex 3
Probes in the fourplex E Sarbeco-FAM Orf1ab S LNA-HEX Spike D614 LNA-TEX B2M-Cy5
Lowest numbers of copies/reaction Test 1 (Ct) Test 2 (Ct) Test 3 (Ct) Test 1 (Ct) Test 2 (Ct) Test 3 (Ct) Test 1 (Ct) Test 2 (Ct) Test 3 (Ct) Test 1 (Ct) Test 2 (Ct) Test 3 (Ct)
1.4 36.96 36.89 36.83 nd nd nd nd nd nd 32.67 32.85 32.51
14 32.14 32.61 32.28 32.71 32.92 32.84 33.77 34.60 34.11 29.31 29.41 29.27
140 28.78 28.91 28.72 30.46 30.38 30.27 31.08 31.14 31.01 26.47 26.49 26.31
1.4E+03 26.11 26.70 26.34 27.92 28.15 28.02 28.71 29.22 28.94 24.37 24.10 24.04
1.4E+04 21.77 21.73 21.59 25.98 26.51 26.12 24.37 24.23 24.18 19.33 19.32 19.22
1.4E+05 17.88 17.59 17.48 21.69 21.65 21.57 20.17 20.01 19.92 16.48 16.55 16.39
1.4E+06 13.77 13.83 13.65 17.64 17.61 17.49 15.91 15.93 15.82 13.85 12.54 12.45
1.4E+07 11.10 10.88 10.77 13.26 13.45 13.31 13.60 13.34 13.41 10.85 10.90 10.70
1.4E+08 7.92 7.89 7.81 10.63 10.41 10.52 10.83 10.70 10.68 7.12 7.07 7.01
1.4E+09 5.20 5.05 5.09 7.20 7.08 7.13 7.31 7.34 7.23 4.51 4.56 4.39
NTC Neg Neg Neg Neg Neg Neg Neg Neg Neg Neg Neg Neg
Fourplex 4
Probes in the fourplex E Sarbeco-FAM Orf1ab L LNA-TEX Orf1ab S LNA-HEX B2M-Cy5
Lowest numbers of copies/reaction Test 1 (Ct) Test 2 (Ct) Test 3 (Ct) Test 1 (Ct) Test 2 (Ct) Test 3 (Ct) Test 1 (Ct) Test 2 (Ct) Test 3 (Ct) Test 1 (Ct) Test 2 (Ct) Test 3 (Ct)
1.4 34.75 35.88 35.70 33.33 34.52 34.47 34.56 33.83 33.97 34.40 34.44 34.57
14 32.44 32.35 32.29 30.53 32.14 32.26 32.76 32.07 32.21 31.63 31.65 31.77
140 29.88 29.82 29.69 27.73 27.76 27.58 29.84 29.93 29.76 29.11 29.01 29.22
1.4E+03 25.17 25.13 25.01 24.97 25.41 25.12 27.48 27.50 27.39 26.52 26.54 26.62
1.4E+04 22.11 22.26 22.02 22.28 22.33 22.19 23.75 23.77 23.64 22.37 22.33 22.47
1.4E+05 18.87 18.54 18.42 19.28 19.72 19.46 20.72 21.07 20.87 18.69 19.13 19.22
1.4E+06 15.92 15.70 15.61 16.35 16.52 16.29 17.66 17.52 17.33 15.37 15.35 15.42
1.4E+07 11.07 10.97 10.88 13.45 13.60 13.32 14.56 14.48 14.39 13.07 12.99 13.21
1.4E+08 7.88 7.94 7.74 8.32 8.34 8.19 8.96 8.99 8.84 9.20 9.22 9.37
1.4E+09 4.91 5.05 4.85 6.44 6.46 6.32 6.42 6.45 6.35 6.86 5.53 6.03
NTC Neg Neg Neg Neg Neg Neg Neg Neg Neg Neg Neg Neg
Fourplex 5
Probes in the fourplex N LSPQ-HEX Spike D614 LNA-FAM Spike 614G LNA-TEX B2M-Cy5
Lowest numbers of copies/reaction Test 1 (Ct) Test 2 (Ct) Test 3 (Ct) Test 1 (Ct) Test 2 (Ct) Test 3 (Ct) Test 1 (Ct) Test 2 (Ct) Test 3 (Ct) Test 1 (Ct) Test 2 (Ct) Test 3 (Ct)
1.4 nd nd nd nd nd nd nd nd nd 33.65 33.85 33.71
14 36.68 36.81 36.72 34.88 35.94 34.76 33.95 34.15 34.03 32.68 32.71 32.59
140 33.20 33.46 33.31 31.70 32.43 32.01 31.72 31.83 31.66 30.52 30.54 30.41
1.4E+03 29.40 29.65 29.52 28.99 29.16 28.89 28.45 28.76 28.37 26.68 26.71 26.52
1.4E+04 24.67 24.75 24.69 25.11 25.46 25.22 24.68 24.88 24.55 23.45 23.48 23.33
1.4E+05 20.66 20.83 20.71 20.37 20.44 20.28 19.82 19.97 19.69 19.32 19.56 19.22
1.4E+06 17.15 17.17 17.05 16.70 16.84 16.62 16.46 16.70 16.39 14.10 14.22 14.02
1.4E+07 14.04 14.00 13.92 12.91 12.95 12.81 12.62 12.65 12.51 11.22 11.39 11.13
1.4E+08 11.15 11.21 11.08 9.58 9.57 9.41 9.29 9.31 9.19 8.41 8.47 8.32
1.4E+09 7.91 7.98 7.84 6.21 6.25 6.11 6.10 6.14 6.02 5.02 5.04 4.94
NTC Neg Neg Neg Neg Neg Neg Neg Neg Neg Neg Neg Neg
Fourplex 6
Probes in the fourplex E Sarbeco-FAM Spike 501Y LNA-HEX Spike 614G LNA-TEX B2M-Cy5
Lowest numbers of copies/reaction Test 1 (Ct) Test 2 (Ct) Test 3 (Ct) Test 1 (Ct) Test 2 (Ct) Test 3 (Ct) Test 1 (Ct) Test 2 (Ct) Test 3 (Ct) Test 1 (Ct) Test 2 (Ct) Test 3 (Ct)
1.4 nd nd 35.64 nd 35.54 35.69 nd nd nd 33.31 33.33 33.29
14 33.71 33.61 33.66 32.49 31.92 32.94 33.77 34.05 33.06 30.94 30.57 30.58
140 30.84 30.85 30.90 29.73 29.70 29.95 31.51 31.11 31.14 28.41 28.48 28.49
1.4E+03 27.64 27.55 27.64 26.63 26.55 26.64 27.91 27.88 27.79 25.43 25.43 25.50
1.4E+04 24.02 23.95 23.95 22.88 22.88 22.87 24.25 24.21 24.27 21.64 21.86 21.60
1.4E+05 20.45 20.45 20.30 19.64 19.67 19.64 20.91 20.94 20.79 18.03 17.62 17.56
1.4E+06 17.12 17.09 16.98 15.88 15.96 15.72 17.19 17.26 16.94 14.39 14.39 14.31
1.4E+07 13.60 13.59 13.38 12.21 12.06 12.09 13.29 13.11 13.18 11.46 11.56 11.40
1.4E+08 9.85 9.74 9.75 9.33 9.28 9.24 10.78 10.77 10.71 6.83 6.40 6.51
1.4E+09 7.01 6.93 6.85 6.91 6.84 6.96 7.65 7.61 7.65 3.61 4.00 3.51
NTC Neg Neg Neg Neg Neg Neg Neg Neg Neg Neg Neg Neg
Fourplex 7
Probes in the fourplex N LSPQ-HEX Spike N501 LNA-FAM Spike 614G LNA-TEX B2M-Cy5
Lowest numbers of copies/reaction Test 1 (Ct) Test 2 (Ct) Test 3 (Ct) Test 1 (Ct) Test 2 (Ct) Test 3 (Ct) Test 1 (Ct) Test 2 (Ct) Test 3 (Ct) Test 1 (Ct) Test 2 (Ct) Test 3 (Ct)
1.4 nd nd nd 35.02 nd nd nd 36.2 nd 33.42 33.52 33.92
14 36.29 36.89 36.71 33.77 34.20 34.92 33.46 33.97 32.80 31.46 31.39 31.32
140 34.27 34.36 34.45 31.18 31.51 31.28 30.72 30.76 30.69 28.77 29.03 28.76
1.4E+04 27.75 27.71 27.75 23.84 24.48 24.34 24.22 24.07 24.17 21.40 21.96 22.24
1.4E+05 24.02 24.12 24.00 20.65 20.55 20.61 20.29 20.52 20.27 18.23 18.17 18.21
1.4E+06 20.56 20.58 20.51 16.73 16.90 16.81 16.60 16.61 16.53 14.21 14.27 14.29
1.4E+07 17.01 17.10 17.05 13.15 13.09 13.05 12.86 12.79 12.85 10.91 11.04 11.09
1.4E+08 13.69 13.66 13.74 9.64 9.24 9.61 10.31 10.26 10.37 7.46 7.40 7.53
1.4E+09 10.10 10.12 10.13 6.49 6.50 6.58 7.53 7.54 7.53 4.46 4.34 4.45
NTC Neg Neg Neg Neg Neg Neg Neg Neg Neg Neg Neg Neg