Skip to main content

Table 7 IFNγ ELISPOT assay precision

From: Validation of a HLA-A2 tetramer flow cytometric method, IFNgamma real time RT-PCR, and IFNgamma ELISPOT for detection of immunologic response to gp100 and MelanA/MART-1 in melanoma patients

(A) Intra assay precision

TIL

TIL1520

TIL1520

TIL1235

     

Peptide

gp100209

gp100pool

MART-1

     

Peptide specificity

Relevant

Relevant

Relevant

     

Test

        

1

34

33

19

     

2

30

43

29

     

3

36

35

26

     

4

40

35

23

     

5

45

41

25

     

6

37

34

21

     

7

32

34

18

     

8

28

35

22

     

Mean (n = 8)

35

36

23

     

SD

5.3

3.6

3.4

     

% CV

15%

10%

15%

     

(B) Inter Assay Precision

Cells

TIL1520

TIL1520

TIL1235

PBMC

PBMC

TIL1235

TIL1235

TIL1520

Peptide

gp100 209

gp100 pool

MART-1

Flu

HIV

gp100 209

gp100 pool

MART-1

Specificity

Relevant

Relevant

Relevant

  

Irrelevant

Irrelevant

Irrelevant

Tests

        

1

44

41

8

195

1

2

2

1

2

42

31

9

206

1

0

1

1

3

37

40

10

198

1

2

1

1

4

43

32

10

222

1

1

1

1

5

38

52

7

240

1

1

1

2

6

38

39

10

254

1

0

2

2

7

39

54

12

278

1

1

1

2

8

49

46

13

245

0

1

1

2

9

36

32

11

226

1

2

1

4

10

38

34

8

224

1

1

1

1

Mean (n = 10)

41

40

10

229

0.9

1.2

1.3

1.6

SD

3.9

8.4

1.8

26.0

0.3

0.8

0.6

0.9

% CV

9.6

20.8

18.3

11.4

33.3

66.6

46.2

56.3

  1. A total of 80 TIL cells were spiked into 105 PBMC per well (Low PBMC) and tested in 8 repeats (intra assay) and 10 tests (inter assay) by two analysts. Average IFNγ gsecreting cells per well (triplicate wells) upon peptide stimulation are shown. Mean, SD, and %CV is shown for PBMC 1 by Analyst 1 as an example.
Back to article page

Journal of Translational Medicine

ISSN: 1479-5876

Contact us