Skip to main content

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