Cross-Platform Evaluation of Commercial Real-Time SYBR Green RT-PCR Kits for Sensitive and Rapid Detection of European Bat Lyssavirus Type 1
Table 2
Comparison of RT-qPCR SybrGreen kits (two-step) provided by the same manufacturer. Results of five different qPCR kits on a range of seven triplicate dilutions of EBLV-1 RNA.
Testing of different RT-qPCR kits (two-step) provided by the same manufacturer
Number of copies/sample
Number of copies/rxn
IScript RT + qPCR SsoFast
QuantiTect RT + qPCR Quantitect
Maxima RT + qPCR Maxima
SS Vilo RT + qPCR Express
qScript RT + qPCR Perfecta
Average Ct
CV (%)
Average Ct
CV (%)
Average Ct
CV (%)
Average Ct
CV (%)
Average Ct
CV (%)
106
5.0 105
16.58 ± 0.13
0.77
19.47 ± 0.07
0.34
16.50 ± 0.15
0.92
15.01 ± 0.05
0.35
16.09 ± 0.03
0.19
18.96 ± 0.04
0.21
22.79 ± 0.26
1.15
17.47 ± 0.09
0.52
17.49 ± 0.10
0.54
17.99 ± 0.29
1.59
105
5.0 104
19.93 ± 0.09
0.45
22.89 ± 0.10
0.42
19.50 ± 0.06
0.33
18.21 ± 0.05
0.26
19.79 ± 0.03
0.13
22.17 ± 0.06
0.25
25.94 ± 0.09
0.35
21.16 ± 0.29
1.38
20.95 ± 0.03
0.15
21.27 ± 0.15
0.72
104
5.0 103
23.28 ± 0.05
0.20
25.89 ± 0.06
0.25
22.95 ± 0.28
1.23
21.68 ± 0.17
0.76
23.18 ± 0.16
0.67
25.75 ± 0.05
0.18
29.60 ± 0.42
1.43
24.29 ± 0.29
0.39
24.40 ± 0.12
0.49
24.55 ± 0.08
0.33
103
5.0 102
26.61 ± 0.21
0.80
29.64 ± 0.32
1.07
26.43 ± 0.26
0.98
24.81 ± 0.05
0.19
26.94 ± 0.08
0.30
29.16 ± 0.23
0.80
32.10 ± 0.19
0.60
27.78 ± 0.30
1.09
28.03 ± 0.74
2.63
27.84 ± 0.12
0.42
102
5.0 101
30.45 ± 0.13
0.42
32.20 ± 0.41
1.26
30.59 ± 0.45
1.46
28.33 ± 0.64
2.25
31.40 ± 0.53
1.68
32.38 ± 0.61
1.89
35.12 ± 0.10
0.28
30.46 ± 0.92
3.03
31.05 ± 0.38
1.23
31.35 ± 0.70
2.22
101
5.0 100
2/3**
1/3**
2/3**
31.52 ± 1.33
4.20
1/3**
1/3**
1/3**
32.76 ± 0.70
2.12
1/3**
2/3**
1
0.5
1/3**
0/3**
0/3**
0/3**
0/3**
0/3**
0/3**
0/3**
0/3**
2/3**
Efficiency
99%
99%
100%
101%
90%
96%
107%
97%
93%
102%
0.999
0.997
0.997
0.999
0.999
0.999
0.991
0.997
0.994
0.998
y-intercept
36.65
39.55
36.31
34.73
37.67
39.38
41.83
38.01
38.50
38.01
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).
