Cross-Platform Evaluation of Commercial Real-Time SYBR Green RT-PCR Kits for Sensitive and Rapid Detection of European Bat Lyssavirus Type 1
Table 3
Comparison of qRT-PCR SYBR Green (one-step) kits. Results of five different qRT-PCR kits on a range of seven triplicate dilutions of EBLV-1 in vitro RNA transcripts.
Testing of different qRT-PCR kits (one-step)
Number of copies/sample
Number of copies/rxn
Quantitect-1 step
Rotor Gene-1 step
Verso-1 step
SSIII Platinium-1 step
qScript-1 step
Average Ct
CV (%)
Average Ct
CV (%)
Average Ct
CV (%)
Average Ct
CV (%)
Average Ct
CV (%)
106
2.0 106
16.66 ± 0.08
0.45
17.15 ± 0.05
0.29
16.99 ± 0.06
0.38
14.52 ± 0.19
0.32
13.47 ± 0.03
0.19
17.79 ± 0.24
1.32
15.88 ± 0.22
1.36
18.34 ± 0.32
1.72
16.22 ± 0.04
0.22
18.48 ± 0.04
0.20
105
2.0 105
20.06 ± 0.10
0.52
20.62 ± 0.04
0.17
20.40 ± 0.11
0.52
17.90 ± 0.21
1.18
16.55 ± 0.03
0.19
21.13 ± 0.45
2.12
19.42 ± 0.20
1.04
21.60 ± 0.15
0.71
19.64 ± 0.41
2.11
22.38 ± 0.15
0.68
104
2.0 104
23.58 ± 0.14
0.58
23.91 ± 0.08
0.33
23.82 ± 0.20
0.83
21.43 ± 0.12
0.57
19.86 ± 0.04
0.18
24.60 ± 0.20
0.83
22.87 ± 0.16
0.69
25.17 ± 0.17
0.69
23.08 ± 0.27
1.16
26.11 ± 0.10
0.38
103
2.0 103
26.70 ± 0.24
0.91
27.45 ± 0.28
1.01
26.93 ± 0.03
0.11
24.90 ± 0.23
0.92
23.13 ± 0.16
0.67
27.95 ± 0.26
0.93
26.34 ± 0.33
1.24
28.63 ± 0.30
1.06
26.14 ± 0.10
0.38
29.54 ± 0.25
0.86
102
2.0 102
30.42 ± 0.08
0.26
2/3**
30.19 ± 0.60
1.98
0/3**
0/3**
31.98 ± 0.17
0.53
29.77 ± 0.39
1.32
31.74 ± 0.79
2.50
28.93 ± 0.08
0.28
31.07 ± 0.37
1.18
101
2.0 101
33.24 ± 0.29
0.87
1/3**
33.27 ± 1.53
4.60
0/3**
0/3**
34.34 ± 1.03
2.99
32.84 ± 0.44
1.33
2/3**
29.91 ± 0.10
0.33
0/3**
1
2
0/3**
1/3**
0/3**
0/3**
0/3**
0/3**
0/3**
2/3**
0/3**
0/3**
Efficiency
98%
96%
100%
94%
104%
97%
94%
95%
100%
87%
0.998
0.999
0.999
0.998
0.999
0.996
0.998
0.997
0.996
0.998
y-intercept
36.89
37.67
36.99
35.29
32.78
38.15
36.80
38.93
38.93
40.74
CV (%) = standard deviation/mean Ct × 100. CV was not calculated when some of the parallels failed to amplify. : number of positives/number of experiments. Average Ct values: mean ± standard deviation. All PCR comparisons were run on the Rotor Gene Q MDx thermocycler (in italic font) and on the Mx3005P thermocycler (in bold).
