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Table 2 Assessment of the reproducibility of the multiplex real-time PCR assay on diluted plasmid DNA containing cloned target sequences

From: The sensitivity of real-time PCR amplification targeting invasive Salmonellaserovars in biological specimens

Amplification target

Result

Target copies

  

5 × 104

5 × 103

5 × 102

5 × 101

5 × 100

S. Typhi without internal control

Ct value*

22.76

25.61

27.92

31.22

-

S. Typhi with internal control

Ct value

22.07

25.56

27.76

31.81

-

Intra-assay variation†

CV (%)

0.45

0.25

0.41

0.67

1.01

Inter-assay variation‡

CV (%)

1.86

1

1.97

3.39

1.19

S. Paratyphi A without internal control

Ct value

21.27

24.78

27.77

31.16

-

S. Paratyphi A with internal control

Ct value

21.42

24.66

28.28

31.84

-

Intra-assay variation

CV (%)

0.41

0.7

0.67

0.86

1.34

Inter-assay variation

CV (%)

1.34

1.34

0.86

1.25

1.83

  1. * Mean Ct value calculated from 4 individual replicates on 4 separate days, n = 16. †Intra-assay variation was calculated by measuring the co-efficient of variance of Ct value on 4 concurrently run assays. ‡ Inter-assay variation was calculated by comparing variation in Ct value on experiments on 4 individual occasions.