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Table 2 Inter-assay variability of IFNγ, IL2 and IFNγ + IL2 responses in the IFNγ/IL2 FluoroSpot assay

From: Validation of an IFNγ/IL2 FluoroSpot assay for clinical trial monitoring

Donor Antigen Plate 1 Plate 2 Plate 1 and plate 2
Mean Mean Mean SD % CV
A
 S09 BZLF1 274 233 254 28.99 11.44
 S11 BZLF1 317 326 322 6.36 1.98
 S12 EBNA3A 92 94 93 1.41 1.52
 S13 EBNA3A 65 63 64 1.41 2.21
 S15 PHA-L 645 680 663 24.75 3.74
 S21 PHA-L 668 753 711 60.10 8.46
 S25 PHA-L 234 287 261 37.48 14.39
 S26 PHA-L 214 240 227 18.38 8.10
Mean      22.36 6.48
B
 S09 BZLF1 56 47 52 6.36 12.36
 S11 BZLF1 52 86 69 24.04 34.84
 S12 EBNA3A 33 35 34 1.41 4.16
 S13 EBNA3A 0 0 0
 S15 PHA-L 575 657 616 57.98 9.41
 S21 PHA-L 698 691 695 4.95 0.71
 S25 PHA-L 411 403 407 5.66 1.39
 S26 PHA-L 547 617 582 49.50 8.50
Mean      21.42 10.20
C
 S09 BZLF1 52 49 51 2.12 4.20
 S11 BZLF1 61 85 73 16.97 23.25
 S12 EBNA3A 19 28 24 6.36 27.08
 S13 EBNA3A 0 0 0
 S15 PHA-L 329 349 339 14.14 4.17
 S21 PHA-L 453 459 456 4.24 0.93
 S25 PHA-L 86 105 96 13.44 14.07
 S26 PHA-L 102 130 116 19.80 17.07
Mean      11.01 12.97
  1. Values represent the mean number of antigen-specific IFNγ (A), IL2 (B) and IFNγ + IL2 (C) SFC/2 × 105 PBMC (stimulated with 1 µg/ml BZLF1 or EBNA3A peptide pools) or mitogen-specific IFNγ (A), IL2 (B) and IFNγ + IL2 (C) SFC/2 × 105 PBMC/5 × 104 PBMC (stimulated with 2 µg/ml PHA-L) detected in two different assay plates performed on the same day in parallel
  2. SD standard deviation, CV coefficient of variation