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Table 5 Repeatability and intermediate precision of real-time RT-PCRs for H1av, H1hu, H1huΔ146–147, H3, N1 and N2 amplifications

From: Molecular subtyping of European swine influenza viruses and scaling to high-throughput analysis

Real-time RT-PCRa for molecular subtyping

RNA extract from (virus strain – name and subtype)

RNA extract dilution (≈ M gene Cq-value)

Mean Cq-value

SD

Inter-assay CV (%)

H1av

A/Sw/Cotes d’Armor/0388/2009 (H1avN1)

10−4 (≈ 23.83)

24.50

0.9

1.20

H1hu

A/Sw/Scotland/410440/1994 (H1huN2)

10−3 (≈ 20.43)

25.63

0.39

1.52

H1hu Δ146–147

A/Sw/France/22–130212/2013 (H1huN2Δ146–147)

10−4 (≈ 21.3)

22.80

0.71

3.12

H3

A/Sw/Flandres/1/1998 (H3N2)

10−2 (≈ 16.7)

18.60

0.31

1.68

N1

A/Sw/Cotes d’Armor/0388/2009 (H1avN1)

10−4 (≈ 23.83)

25.64

0.37

1.46

N2

A/Sw/Scotland/410440/1994 (H1huN2)

10−3 (≈ 20.43)

24.78

1.19

4.79

A/Sw/Flandres/1/1998 (H3N2)

10−3 (≈ 20.21)*

28.56

1.86

6.51

  1. aHA RT-qPCRs were run as simplex assays; N1 and N2 RT-qPCRs were run in duplex. Cq quantification threshold, SD standard deviation, CV coefficient of variation. Each RNA extract was tested (either on Thermocycler MxPro – Mx3005P or Chromo4) in 6 different assays at the dilution corresponding to 10 times the detection limit of the concerned RT-qPCR (except H3 that was run on a lowest dilution of the H3N2 strain). The RNA extract dilution marked with an asterisk (*) was tested in 5 independent assays