4th ISTA Proficiency Test on GMO Testing on Soybean
ISTA Proficiency Working Groupof the GMO TF
B. Kahlert, S. Grégoire, C. Haldemann
ISTA Ordinary Meeting – Results of the ISTA GMO Proificiency Tests
Outline
• IntroductionDesign of the 4th Proficiency TestWhat is new in this test round?
• ResultsQualitative TestSub-sampling QuantificationQuantitative Test
• Summary
ISTA Ordinary Meeting – Results of the ISTA GMO Proificiency Tests
Design of the 4th Proficiency Test
• crop: Glycine max (L.) Merr.• # of samples per lab: 12• # of negative samples: 3• # of positive samples: 9
0.1%: 30.5%: 31.0%: 3
• event: GTS-40-3-2• sample size: 3000 seeds• qualitative, sub-sampling quantification and
quantitative test
ISTA Ordinary Meeting – Results of the ISTA GMO Proificiency Tests
What was new in this test round?
• crop: soybean• event was known by the lab• information about source of reference material was
provided• seeds for each sample were counted• conventional seed and GM seed part were weighted • Qualitative and quantitative test were obligatory
ISTA Ordinary Meeting – Results of the ISTA GMO Proificiency Tests
Submitted Test Results
qual. & sub-sampl.
11 (18%)
qual., sub-sampl. & quant.
1 (2%)
qual.3 (5%)
no results9 (15%)
qual. & quant.36 (60%)
ISTA Ordinary Meeting – Results of the ISTA GMO Proificiency Tests
Performed tests by the laboratories...
PT01 PT02 PT03 PT04no results 11% 10% 17% 15%qual. 89% 29% 23% 5%qual. & sub-sampl. - 23% 15% 18%qual. & quant. - 30% 45% 60%qual. & sub-sampl. & quant. - 8% 0% 2%
Total # of laboratories 43 52 48 60
Qualitative Test
Qualitative Result:
• The detection of GM seeds in conventional seed lots.
• The laboratory classified each of the 12 samples as either positive or negative.
ISTA Ordinary Meeting – Results of the ISTA GMO Proficiency Tests
Qualitative Test
correct results42 (82%)
false results9 (18%) false negative
results5 (10%)
false positive results3 (6%)
both false neg. and
pos. results 1 (2%)
ISTA Ordinary Meeting – Results of the ISTA GMO Proficiency Tests
Laboratories which reported...
PT01 PT02 PT03 PT04Correct results 70% 85% 81% 82%False results 30% 15% 19% 18%False negative 19% 4% 12% 12%False positive 7% 4% 5% 8%Both false pos & neg 5% 6% 2% 2%
Qualitative Test
153
153
153
459
153
612
3
0
7
10
8
18
0% 20% 40% 60% 80% 100%
1.0% GM seeds
0.5% GM seeds
0.1% GM seeds
Positive samples
Negative samples
All samples
% of samples tested# of samples reported as false
2.9%5.2%2.2%4.6%0.0%2.0%
ISTA Ordinary Meeting – Results of the ISTA GMO Proficiency Tests
Samples classified as...
PT01 PT02 PT03 PT04Correctly reported results 95.4% 96.4% 95.9% 97.1%False negative 6.5% 3.0% 4.3% 2.2%False positive 1.7% 5.0% 3.3% 5.2%
Sub-sampling Quantification
The laboratory reported the test plan...• the size of sub-samples (in number of seeds)• the number of sub-samples tested• the number of positive sub-samples
and the results...• whether the % of GM seeds is ≤ 1% or >1%• the estimate of the percentage of GM seeds per
sample (in number of seeds)
ISTA Ordinary Meeting – Results of the ISTA GMO Proficiency Tests
Sub-sampling Quantification
Tools: Seedcalc6 and % gm estimate.xls
ISTA Ordinary Meeting – Results of the ISTA GMO Proficiency Tests
Sub-sampling QuantificationISTA Ordinary Meeting – Results of the ISTA GMO Proficiency Tests
Laboratory’s estimated results and its mean
Labs# of sub-samples
size ofsub-sample
(1), 2, 3, 4 30 1005, 6 20 150
7 15 2008, 9 10 300
10 6 50011 2 to 6 100 to 300
Estim
ate
of %
GM
see
ds
0
0.2
0.4
0.6
0.8
1
1.2
1.4
lab1
lab2
lab3
lab4
lab5
lab6
lab7
lab8
lab9
lab1
0
lab1
1
0
0.2
0.4
0.6
0.8
1
1.2
1.4
lab1
lab2
lab3
lab4
lab5
lab6
lab7
lab8
lab9
lab1
0
lab1
1
0
0.2
0.4
0.6
0.8
1
1.2
1.4
lab1
lab2
lab3
lab4
lab5
lab6
lab7
lab8
lab9
lab1
0
lab1
1
Spiking level 0.1% Spiking level 0.5% Spiking level 1.0%
Sub-sampling Quantification
Spiking level
0.1% 0.5% 1.0%
Test
resu
lts (%
)
0.0
0.2
0.4
0.6
0.8
1.0
1.2
1.4
ISTA Ordinary Meeting – Results of the ISTA GMO Proficiency Tests
and PT03 (5 labs)
Spiking level
0.2% 2.0% 4.0%
Test
resu
lts (%
)0.0
1.0
2.0
3.0
4.0
5.0
Results of PT04 (11 labs)
Quantitative Test
Quantitative Result:
• The laboratory reported the GM content of thepositive test samples.
• The result should be given in % of GM seed by mass or % seed by number.
• The result could be given in another unit chosen bythe laboratory.
36 laboratories reported in 39 test series
ISTA Ordinary Meeting – Results of the ISTA GMO Proficiency Tests
Quantitative TestISTA Ordinary Meeting – Results of the ISTA GMO Proficiency Tests
Protocol of Results
ISTA Ordinary Meeting – Results of the ISTA GMO Proficiency Tests
Quantitative Test
Spiking level 0.1%
0.0
0.2
0.4
0.6
0.8
1.0
all
lab1
2
lab1
3
lab1
4
lab1
5
lab1
6
lab1
7
lab1
8
lab1
9
lab2
0
lab2
1
lab2
2
lab2
3
lab2
4
lab2
5
lab2
6
lab2
7
lab2
8
lab2
9
lab3
0
lab3
1
lab3
2
lab3
3
lab3
4
lab3
5
lab3
6a
lab3
7
lab3
8
lab3
9
lab4
0
lab4
1
lab4
2
lab4
3a
lab4
4
lab4
5
lab4
6
lab4
7
lab3
6b
lab4
3b
lab4
8
Mea
n pe
rcen
tage
GM
see
ds +
/- SD
(%)
NB : laboratory number has been re-coded
ISTA Ordinary Meeting – Results of the ISTA GMO Proficiency Tests
Quantitative Test
Spiking level 0.5%
0.0
0.2
0.4
0.6
0.8
1.0
1.2
1.4
1.6
1.8
2.0
2.2
2.4
all
lab2
6
lab1
2
lab1
3
lab2
2
lab2
1
lab1
4
lab2
3
lab2
4
lab4
2
lab4
7
lab2
9
lab1
5
lab3
0
lab2
7
lab1
6
lab3
6a
lab3
1
lab1
7
lab1
8
lab3
9
lab3
4
lab3
3
lab2
8
lab1
9
lab4
1
lab4
0
lab3
5
lab4
6
lab2
5
lab3
7
lab3
8
lab3
6b
lab4
8
lab2
0
lab4
4
lab4
3b
lab3
2
lab4
5
lab4
3a
Mea
n pe
rcen
tage
GM
see
ds +
/- SD
(%)
ISTA Ordinary Meeting – Results of the ISTA GMO Proficiency Tests
Quantitative Test
Spiking level 1.0%
0.00.20.40.60.81.01.21.41.61.82.02.22.42.62.83.03.23.43.63.84.04.2
all
lab1
2
lab1
3
lab2
6
lab2
1
lab3
2
lab2
3
lab2
2
lab1
5
lab4
2
lab2
4
lab4
7
lab2
8
lab1
7
lab3
0
lab1
4
lab3
6a
lab2
5
lab3
3
lab3
9
lab3
5
lab1
9
lab2
0
lab1
6
lab3
7
lab4
1
lab3
8
lab2
7
lab4
0
lab3
1
lab1
8
lab4
4
lab4
8
lab4
6
lab3
6b
lab4
5
lab2
9
lab3
4
lab4
3b
lab4
3a
Mea
n pe
rcen
tage
GM
see
ds +
/- SD
(%)
ISTA Ordinary Meeting – Results of the ISTA GMO Proficiency Tests
Quantitative Test
0.0
0.5
1.0
1.5
2.0
2.5
3.0
lab1
2
lab1
3
lab1
4
lab1
5
lab1
6
lab1
7
lab1
8
lab1
9
lab2
0
lab2
1
lab2
2
lab2
3
lab2
4
lab2
5
lab2
6
lab2
7
lab2
8
lab2
9
lab3
0
lab3
1
lab3
2
lab3
3
lab3
4
lab3
5
lab3
6a
lab3
6b
lab3
7
lab3
8
lab3
9
lab4
0
lab4
1
lab4
2
lab4
3a
lab4
3b
lab4
4
lab4
5
lab4
6
lab4
7
lab4
8
0.1% 0.5% 1.0%
Mean per GMO spiking level and laboratory
Results of PT04
TestSpiking level (%) Mean (%)
Standarddeviation
Variationcoeff.
PT04 0.1 0.13 0.11 87%PT04 0.5 0.47 0.29 61%PT04 1.0 0.87 0.32 37%PT03 0.2 0.19 0.10 51%PT03 2.0 1.62 0.92 57%PT03 4.0 3.37 1.68 50%PT02 0.7 0.73 0.37 50%PT02 1.4 1.47 0.91 62%
Spiking level
0.1% 0.5% 1.0%
Test
resu
lts (%
)
0.0
0.5
1.0
1.5
2.0
2.5
3.0
ISTA Ordinary Meeting – Results of the ISTA GMO Proficiency Tests
Quantitative Test
ISTA Ordinary Meeting – Results of the ISTA GMO Proficiency Tests
Summary/ previous testsPT04 - Qualitative Results:• The percentage of laboratories reporting correct
results for all samples is similar to the previous tests• >80% of labs made no missclassification, • >95% of samples are reported correctlyPT04 - Quantitative Results:• The results show variation increasing along with the
spiking level as in other tests.• A higher number of laboratories reported results that
were close to the true value (spiking level). Theinformation about event and the availability of reference material might have helped the laboratoriesto achieve better results in this round.
ISTA Ordinary Meeting – Results of the ISTA GMO Proficiency Tests
5th ISTA Proficiency Teston GMO Testing
• Crop: Soybean• 12 samples of 3000 seeds• Qualitative Test• Sub-sampling quantification or quantitative test• Material Transfer Agreement
• Register now!
ISTA Ordinary Meeting – Results of the ISTA GMO Proficiency Tests
& Registration FormAnnouncement