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Table 3 Analytical sensitivity of the real-time RT-PCRs for H1av, H1hu, H3, N1 and N2 on ten-fold dilutions of RNA extracted from swIAVs of different subtypes

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

Virus strain Virus stock dilution M gene RT-qPCRa(Cq-value) Real-time RT-PCRb for molecular subtyping (Cq-value)
Lineage Name
H1av H1hu H1huΔ146–147 H3 N1 N2
H1avN1 A/Sw/Cotes d’Armor/0388/2009 10−1 12.52 16.10 no Cq nt no Cq 15.52 no Cq
10−2 16.09 19.52 no Cq nt no Cq 18.75 no Cq
10−3 20.16 22.77 no Cq nt no Cq 22.46 no Cq
10−4 23.83 25.85 no Cq nt no Cq 25.71 no Cq
10−5 27.03 28.86 no Cq nt no Cq 28.18 no Cq
10−6 29.79 32.32 no Cq nt no Cq no Cq no Cq
10−7 33.36 36.26 no Cq nt no Cq no Cq no Cq
10−8 38.59 no Cq no Cq nt no Cq no Cq no Cq
10−9 no Cq no Cq no Cq nt no Cq no Cq no Cq
10−10 no Cq no Cq no Cq nt no Cq no Cq no Cq
H1huN2 A/Sw/Scotland/410440/1994 10−1 13.47 no Cq 15.18 nt no Cq no Cq 16.90
10−2 17.15 no Cq 18.53 nt no Cq no Cq 19.92
10−3 20.43 no Cq 21.99 nt no Cq no Cq 23.54
10−4 23.79 no Cq 25.19 nt no Cq no Cq 26.60
10−5 27.25 no Cq no Cq nt no Cq no Cq no Cq
10−6 30.11 no Cq no Cq nt no Cq no Cq no Cq
10−7 33.05 no Cq no Cq nt no Cq no Cq no Cq
10−8 37.48 no Cq no Cq nt no Cq no Cq no Cq
10−9 no Cq no Cq no Cq nt no Cq no Cq no Cq
10−10 no Cq no Cq no Cq nt no Cq no Cq no Cq
H1huN2Δ146–147 A/Sw/France/22–130212/2013 10−1 10.98 nt 11.88 12.68 nt no Cq 11.48
10−2 14.67 nt 15.26 16.10 nt no Cq 14.38
10−3 17.82 nt 18.66 20.07 nt no Cq 17.78
10−4 21.3 nt 21.77 22.95 nt no Cq 20.92
10−5 25.64 nt 25.03 26.53 nt no Cq 24.54
10−6 28.06 nt no Cq 29.83 nt no Cq 33.03*
10−7 31.54 nt no Cq 33.36 nt no Cq no Cq
10−8 35.16 nt no Cq 37.03 nt no Cq no Cq
10−9 no Cq nt no Cq 39.63 nt no Cq no Cq
10−10 no Cq nt no Cq no Cq nt no Cq no Cq
H3N2 A/Sw/Flandres/1/1998 10−1 13.13 no Cq no Cq nt 17.045 no Cq 16.865
10−2 16.7 no Cq no Cq nt 19.88 no Cq 19.725
10−3 20.21 no Cq no Cq nt 23.55 no Cq 23.275
10−4 23.57 no Cq no Cq nt 27.075 no Cq 26.67*
10−5 26.9 no Cq no Cq nt 30.235 no Cq no Cq
10−6 30.07 no Cq no Cq nt 33.53 no Cq no Cq
10−7 32.7 no Cq no Cq nt 36.335 no Cq no Cq
10−8 37.04 no Cq no Cq nt 37.86 no Cq no Cq
10−9 no Cq no Cq no Cq nt 39.99 no Cq no Cq
10−10 no Cq no Cq no Cq nt no Cq no Cq no Cq
  1. Cq quantification cycle, nt not tested. M gene RT-qPCRs were run on one replicate whereas molecular subtyping RT-qPCRs were run on duplicates (Thermocycler MxPro – Mx3005P). Thus, Cq-values for molecular subtyping RT-qPCRs are mean Cq-values between duplicates, except numbers indicated in italics with an asterisk (*). Cq-values indicated in bold represent detection limits. aCommercial kits [21]; bHA RT-qPCRs were run as simplex assays; N1 and N2 RT-qPCRs were run in duplex