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August 2015 DLA – 24/2015 – GMO-Screening qualitative
DLADienstleistung
LebensmittelAnalytik GbR
Evaluation Reportproficiency test
24/2015
GMO - Screening qualitative:
5 Samples with positive/negative amounts of GMO-Maize (Bt11) or GMO-Soya (RR)
Dienstleistung Lebensmittel Analytik GbRWaldemar-Bonsels-Weg 17022926 Ahrensburg, Germany
[email protected] www.dla-lvu.de
Coordinator of this PT: Dr. Matthias Besler
Reprint, also in part, only with written permission from DLA-AhrensburgPage 1 of 25
August 2015 DLA – 24/2015 – GMO-Screening qualitative
Content1. Introduction..................................................32. Realisation...................................................3
2.1 Test material............................................32.2 Test.....................................................32.3 Submission of results....................................3
3. Evaluation....................................................54. Results.......................................................5
4.1 Test.....................................................64.1.1 Results: 35S-Screening-Sequence........................64.1.2 Results: NOS-Screening-Sequence........................74.1.3 Results: GMO-Soya (RR-Round-Up-Ready-Soya).............84.1.4 Results: Lectin-DNA (Soya-specific)....................94.1.5 Results: GMO-Maize (bt11-Maize).......................104.1.6 Results: Maize-DNA (Maize-specific)...................114.1.7 Results: Other Parameters (DNA).......................12
5. Documentation................................................135.1 Details by participants about DNA-Extraction methods. . . .135.1.1 35S-Screening Sequence................................135.1.2 NOS-Screening Sequence................................145.1.3 GMO-Soya (RR-Round-Up-Ready-Soya).....................155.1.4 Lectin-DNA (Soya-specific)............................155.1.5 GMO-Maize (bt11-Maize)................................165.1.6 Maize-DNA (Maize-specific)............................165.1.7 Other Parameters (DNA)................................175.2 Details by participants to PCR-reaction.................185.2.1 35S-Screening Sequence................................185.2.2 NOS-Screening Sequence................................195.2.3 GMO-Soya (RR-Round-Up-Ready-Soya).....................205.2.4 Lectin-DNA (Soya-specific)............................215.2.5 GMO-Maize (bt11-Maize)................................225.2.6 Maize-DNA (Maize-specific)............................225.2.7 Other Parameters (DNA)................................23
6. Index of participant laboratories............................247. Index of references..........................................25
Reprint, also in part, only with written permission from DLA-AhrensburgPage 2 of 25
August 2015 DLA – 24/2015 – GMO-Screening qualitative
1. Introduction
The participation in proficiency testing schemes is an essential elementof the quality-management-system of every laboratory testing food andfeed, cosmetics and food contact materials. The implementation ofproficiency tests enables the participating laboratories to prove theirown analytical competence under realistic conditions. At the same timethey receive valuable data regarding the validity of the particulartesting method. The purpose of DLA is to offer proficiency tests for selected parametersin concentrations with practical relevance.Realisation and evaluation of the present proficiency test follows thetechnical requirements of DIN EN ISO/IEC 17043 (2010) and DIN ISO13528:2009.
2. Realisation
2.1 Test material
The test materials are 5 different mixtures of common in commerce foodsfrom European and US-American suppliers (s. table 1).The ingredients were mixed, homogenized and portioned to approximately10 g. The materials were tested for homogeneity.
2.2 Test
One portion of each of the 5 test materials was sent to every participa-ting laboratory in the 21st week of 2015. The testing method was optio-nal. The tests should be finished at July 3rd 2015 the latest.
2.3 Submission of results
The participants submitted their results in standard forms, which havebeen handed out along with the samples. The results given aspositive/negative were evaluated with respect to each tested parameter.Queried and documented were the indicated results and details of the testmethods like specifity, test kit manufacturer and hints about the pro-cedure.All participants submitted their results in time.
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August 2015 DLA – 24/2015 – GMO-Screening qualitative
Table 1: Composition of DLA-samples
DLA-Sample
Ingredients (per 100 g) GMO-Con-tent Maize
GMO-Con-tent Soya
1 Maize Semolina, European-Supplier (62,5 g)Ingredients: Maize FlourNutrients per 100 g: Protein 7,5 g, Carbohydrates 74 g, Fat 1 g
Potato Flour (37,5 g)Ingredients: Potato flourIngredients per 100 g: Protein 0,6 g, Carbohydrates 83 g, Fat 0,1 g
-
-
-
-
2 Maize Semolina, European-Supplier (50 g)Ingredients: Maize FlourNutrients per 100 g: Protein 7,5 g, Carbohydrates 74 g, Fat 1 g
Potato Flour (37,5 g)Ingredients: Potato flourIngredients per 100 g: Protein 0,6 g, Carbohydrates 83 g, Fat 0,1 g
Maize Flour, USA-Supplier (12,5 g)Ingredients: Maize FlourNutrients per 100 g: Protein 9 g, Carbohydrates 79 g, Fat 0 g
-
-
positive(bt11-Maize experimental)
-
-
-
3 Soy Flakes, European Supplier (62 g)Ingredients: SoybeansNutrients per 100 g: Protein 41 g, Carbohydrates 3,1 g, Fatt 21 g
Glucose (19 g)Lactose (19 g)
-
- -
-
- -
4 Meal Replacement, Dietetic Food (62,5 g)Ingredients: Soyprotein isolate, honey, skimmed milk – yoghurt powder, iron-fumarate, zinc oxide, copper gluconate, manganese sulfate, potassium iodide, vitamin A, vitamin B1, vitamin B2, vitamin B6, vitamin B12, biotin, calcium pantothenate, folic acid, nicotinamide, vitamin D3, dl-alpha-toco-pheryl acetate
Potato Flour (37,5 g)Ingredients: Potato flourIngredients per 100 g: Protein 0,6 g, Carbohydrates 83 g, Fat 0,1 g
-
-
positive(RR-Soya experimental)
-
5 Soy Flakes, European Supplier (59 g)Ingredients: SoybeansNutrients per 100 g: Protein 41 g, Carbohydrates 3,1 g, Fatt 21 g
Glucose (18 g)Lactose (18 g)
Soya Chunks, USA-Supplier (5,4 g)Ingredients: Soybean FlourNutrients per 100 g: Protein 47 g, Carbohydrates 17 g, Fat 0,8 g
-
- -
-
-
- -
positive(RR-Soya experimental)
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August 2015 DLA – 24/2015 – GMO-Screening qualitative
3. Evaluation
The evaluation of the GMO-screening proficiency test was done exclusivelyqualitative.
The results are presented for all 5 test samples in separate tables foreach parameter 35S, NOS, GMO-Soya (RR), Lectin-DNA, GMO-Maize (bt11),Maize-DNA and other DNA results.The numbers and percentage of positive and negative results are given atthe end of each table. The participating laboratories decide according totheir own criteria wether a result is valuated positive or negative.If there are ≥ 75 % positive or negative results, a consensus result isdetermined for each sample.
For every participant a qualitative valuation is made with respect to theconsensus results. Therefore the number and percentage of "correct"results of consensus results is given.
4. Results
All following tables are anonymized. With the delivering of the evalua-tion-report the participants are informed about their individual evalua-tion-number.
The results of the participants are given in tables as indicated below:
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Sample 1 Sample 2 Sample 3 Sample 4 Sample 4
Parameter
Evaluation number
Qualitative Valuation
Remarks
pos/neg pos/neg pos/neg pos/neg pos/neg Agreements with consensus results
August 2015 DLA – 24/2015 – GMO-Screening qualitative
4.1 Test
4.1.1 Results: 35S-Screening-Sequence
Comments on results:
For 4 samples consensus results with two times 100%, one time 90% and onetime 86% positive or negative results were obtained.For sample 3 there were 71% negative and 29% positive results. So aconsensus value with ≥ 75% could not be determined. Therefore sample 3was not considered for the "qualitative valuation".
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Sample 1 Sample 2 Sample 3 Sample 4 Sample 5
35S
1 positive positive negative positive positive 3/4 (75%)
2 negative positive positive positive positive 4/4 (100%)
3 negative positive negative positive positive 4/4 (100%)
4 positive positive positive positive positive 3/4 (75%)
5 negative positive positive positive positive 4/4 (100%)
6 negative positive negative positive positive 4/4 (100%)
7 negative positive negative positive positive 4/4 (100%)
8 negative positive negative positive positive 4/4 (100%)
9 negative positive positive positive positive 4/4 (100%)
10 negative positive negative positive positive 4/4 (100%)
11 negative positive negative positive positive 4/4 (100%)
12 negative positive negative positive positive 4/4 (100%)
14 negative positive negative positive positive 4/4 (100%)
15 negative positive negative negative positive 3/4 (75%)
16 negative positive negative negative positive 3/4 (75%)
18 negative positive negative positive positive 4/4 (100%)
19 positive positive negative positive positive 3/4 (75%)
20 negative positive positive positive positive 4/4 (100%)
21 negative positive negative positive positive 4/4 (100%)
22 negative positive positive positive positive 4/4 (100%)
24 negative positive negative positive positive 4/4 (100%)
Sample 1 Sample 2 Sample 3 Sample 4 Sample 53 21 6 19 2118 0 15 2 014 100 29 90 10086 0 71 10 0
negative positive positive positive
Evaluation number
Qualitative Valuation
Remarks
pos/neg pos/neg pos/neg pos/neg pos/negAgreements with consensus value
Sample 3: tracesbelow
absolute LOD
Sample 3: traces ct 38,5
Number positive
Number negative
Percent positive
Percent negative
Consensus none
August 2015 DLA – 24/2015 – GMO-Screening qualitative
4.1.2 Results: NOS-Screening-Sequence
Comments on results:
For all 5 samples consensus results with two times 100% and one time 90%,86% and 81% each positive or negative results were obtained.
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Sample 1 Sample 2 Sample 3 Sample 4 Sample 5
NOS
1 positive positive positive positive positive 3/5 (60%)
2 negative positive negative negative positive 4/5 (80%)
3 negative positive negative positive positive 5/5 (100%)
4 negative positive negative positive positive 5/5 (100%)
5 negative positive positive positive positive 4/5 (80%)
6 negative positive negative positive positive 5/5 (100%)
7 negative positive negative positive positive 5/5 (100%)
8 negative positive negative positive positive 5/5 (100%)
9 negative positive positive positive positive 4/5 (80%)
10 negative positive negative positive positive 5/5 (100%)
11 negative positive negative positive positive 5/5 (100%)
12 negative positive negative positive positive 5/5 (100%)
14 negative positive negative positive positive 5/5 (100%)
15 negative positive negative negative positive 4/5 (80%)
16 negative positive negative negative positive 4/5 (80%)
18 negative positive negative positive positive 5/5 (100%)
19 positive positive negative positive positive 4/5 (80%)
20 negative positive negative positive positive 5/5 (100%)
21 negative positive negative positive positive 5/5 (100%)
22 negative positive positive positive positive 4/5 (80%)
24 negative positive negative positive positive 5/5 (100%)
Sample 1 Sample 2 Sample 3 Sample 4 Sample 52 21 4 18 2119 0 17 3 010 100 19 86 10090 0 81 14 0
negative positive negative positive positive
Evaluation number
Qualitative Valuation
Remarks
pos/neg pos/neg pos/neg pos/neg pos/negAgreements with con-
sensus value
Sample 3: traces below absolute
LOD
* Sample 3: traces ct 38,8
Number positive
Number negative
Percent positive
Percent negative
Consensus
August 2015 DLA – 24/2015 – GMO-Screening qualitative
4.1.3 Results: GMO-Soya (RR-Round-Up-Ready-Soya)
Comments on results:
For 4 samples consensus results with one time 100%, two times 86% and onetime 79% positive or negative results were obtained.For sample 4 there were 71% positive and 29% negative results. So aconsensus value with ≥ 75% could not be determined. Therefore sample 4was not considered for the "qualitative valuation".
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Sample 1 Sample 2 Sample 3 Sample 4 Sample 5
1 positive negative positive positive positive 2/4 (50%)
2 negative negative negative negative positive 4/4 (100%)
3 negative negative negative positive positive 4/4 (100%)
5 negative negative positive positive positive 3/4 (75%)
6 negative negative negative positive positive 4/4 (100%)
11 negative negative negative positive positive 4/4 (100%)
12 negative negative negative positive positive 4/4 (100%)
13 negative negative positive positive positive 3/4 (75%)
15 negative positive negative negative positive 3/4 (75%)
17 negative negative negative positive positive 4/4 (100%)
18 negative negative negative negative positive 4/4 (100%)
19 positive positive negative positive positive 2/4 (50%)
21 negative negative negative positive positive 4/4 (100%)
23 negative negative negative negative positive 4/4 (100%)
Sample 1 Sample 2 Sample 3 Sample 4 Sample 52 2 3 10 1412 12 11 4 014 14 21 71 10086 86 79 29 0
negative negative negative positive
Evaluation number
Qualitative Valuation
Remarks
RR-Soya pos/neg pos/neg pos/neg pos/neg pos/negAgreements with con-
sensus value
Sample 3: traces below relative LOD
Number positive
Number negative
Percent positive
Percent negative
Consensus none
August 2015 DLA – 24/2015 – GMO-Screening qualitative
4.1.4 Results: Lectin-DNA (Soya-specific)
Comments on results:
For 3 samples consensus results with 100% each positive or negativeresults were obtained.For samples 1 and 2 there were 67% negative results. Therefore noconsensus value could be determined and the samples were not included forthe "qualitative valuation".
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Sample 1 Sample 2 Sample 3 Sample 4 Sample 5
Lectin
1 positive positive positive positive positive 3/3 (100%)
2 negative negative positive positive positive 3/3 (100%)
3 negative negative positive positive positive 3/3 (100%)
5 negative positive positive positive positive 3/3 (100%)
6 negative negative positive positive positive 3/3 (100%)
11 negative negative positive positive positive 3/3 (100%)
12 negative negative positive positive positive 3/3 (100%)
13 negative negative positive positive positive 3/3 (100%)
17 positive positive positive positive positive 3/3 (100%)
18 negative negative positive positive positive 3/3 (100%)
19 positive positive positive positive positive 3/3 (100%)
21 positive negative positive positive positive 3/3 (100%)
Sample 1 Sample 2 Sample 3 Sample 4 Sample 54 4 12 12 128 8 0 0 033 33 100 100 10067 67 0 0 0
positive positive positive
Evaluation number
Qualitative Valuation
Remarks
pos/neg pos/neg pos/neg pos/neg pos/neg Agreements with consensus value
Sample 1 + 2 late amplification curve, not
clearly positive
Sample 1: few traces
Number positive
Number negative
Percent positive
Percent negative
Consensus none none
August 2015 DLA – 24/2015 – GMO-Screening qualitative
4.1.5 Results: GMO-Maize (bt11-Maize)
Comments on results:
For all 5 samples consensus results with three times 100% and two times90% positive or negative results were obtained.
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Sample 1 Sample 2 Sample 3 Sample 4 Sample 5
1 negative positive negative positive negative 4/5 (80%)
3 negative positive negative negative negative 5/5 (100%)
5 negative positive negative negative negative 5/5 (100%)
6 negative positive negative negative negative 5/5 (100%)
13 negative positive negative negative negative 5/5 (100%)
15 negative positive negative negative positive 4/5 (80%)
17 negative positive negative negative negative 5/5 (100%)
18 negative positive negative negative negative 5/5 (100%)
19 negative positive negative negative negative 5/5 (100%)
21 negative positive negative negative negative 5/5 (100%)
Sample 1 Sample 2 Sample 3 Sample 4 Sample 50 10 0 1 110 0 10 9 9
Prozent positive 0 100 0 10 10Prozent negative 100 0 100 90 90
negative positive negative negative negative
Evaluation number
Qualitative Valuation
Remarks
bt11 Maize pos/neg pos/neg pos/neg pos/neg pos/neg Agreements with consensus value
Number positive
Number negative
Consensus
August 2015 DLA – 24/2015 – GMO-Screening qualitative
4.1.6 Results: Maize-DNA (Maize-specific)
Comments on results:
For 3 samples consensus results with two times 92% and one time 83%positive results were obtained.For samples 3 and 5 there were 50% positive and 50% negative results.Therefore no consensus value of ≥ 75% could determined for samples 3 and5 and they were not included for the "qualitative valuation". Accordingto participants' remarks only „traces“ of maize DNA were detected insamples 3 and 5.
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Sample 1 Sample 2 Sample 3 Sample 4 Sample 5
1 positive positive positive positive positive 3/3 (100%)3 positive positive negative positive positive 3/3 (100%)5 positive positive positive positive positive 3/3 (100%)6 positive positive negative negative negative 2/3 (100%)
11 positive positive positive positive positive 3/3 (100%)
12 positive positive negative positive negative 3/3 (100%)
13 positive positive positive positive positive 3/3 (100%)15 negative negative negative negative negative 0/3 (0%)17 positive positive positive positive positive 3/3 (100%)18 positive positive negative positive negative 3/3 (100%)19 positive positive negative positive negative 3/3 (100%)
21 positive positive positive positive negative 3/3 (100%)
Sample 1 Sample 2 Sample 3 Sample 4 Sample 511 11 6 10 61 1 6 2 692 92 50 83 508 8 50 17 50
positive positive positive
Evaluation number
Qualitative Valuation
Remarks
Maize DNA pos/neg pos/neg pos/neg pos/neg pos/negAgreements with consen-
sus value
Sample 3 + 5: low content of Maize DNA
Sample 3 + 5: traces;Sample 4: clear traces
Samplen 3+4: low traces
Number positive
Number negative
Percent positive
Percent negative
Consensus none none
August 2015 DLA – 24/2015 – GMO-Screening qualitative
4.1.7 Results: Other Parameters (DNA)
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Parameter Sample 1 Sample 2 Sample 3 Sample 4 Sample 5
1a NK603 positive positive negative positive positive1b negative positive negative positive positive1c bar negative negative negative negative negative1d FMV negative negative negative negative negative1e EPSPS positive negative positive positive2 Chloroplast-DNA positive positive positive positive positive4 FMV negative negative negative negative positive5 FMV negative negative negative negative positive7 FMV negative negative negative negative positive8 negative negative negative negative negative
11 negative positive negative positive positive
12a p35S-pat negative positive negative negative negative12b p35S-pat negative positive negative negative negative
12c negative positive negative positive positive
12d bar negative negative negative negative negative12e negative negative negative negative negative14 FMV negative negative negative positive positive15 negative negative negative negative negative20 Plant-DNA positive positive positive positive positive
21a negative positive negative positive positive
21b negative positive negative positive positive
21c Screening bar negative negative negative negative negative21d Screening FMV negative negative negative negative positive
21e negative negative negative negative positive
24 FMV negative negative negative negative positive
Evaluation number
Remarks
Other DNA pos/neg pos/neg pos/neg pos/neg pos/neg
pat
pos / neg
other
CTP2-CP4-EPSPS
CTP2-CP4-EPSPS
Samples 4 + 5: clear traces
pNOS
Target-DNA
Screening Pat-Gen
Screening CTP2-CP4-EPSPS
GVO-Soja MON89788
Sample 4: traces
August 2015 DLA – 24/2015 – GMO-Screening qualitative
5. Documentation
5.1 Details by participants about DNA-Extraction methods
5.1.1 35S-Screening Sequence
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35S
12
3 §64 LFGB
45
6
7
8
9
10
11 ASU L 00.00-122
12 p35S-spezifische DNA-Sequenzen ASU L 00.00-122
14 35S1516 35 S, NOS
18 P-35S
1920 ASU L00.00.31
21 35S-promoter
22 GEN-IAL24 35S
Evaluation number
Result given as Test-Kit or Literature Remarks to DNA-Extraction
Target-Sequenz / -DNA Supplier / Method e.g. Extraction / Enzymes / Clean-Up / DNA-Quality
extract. 2g with CTAB and clean-up with QIAamp® DNA Stool Mini Kit (Fa. Qiagen)
combination: CTAB-Lysis with Macherey Nagel NucleoSpin® Food Kit
35S CaMV-Promotor SureFood GMO Kit, R-Biopharm NucleoSpin Food, Fa. Machery & NagelGen-ial FFS-Kit, Promega
detected or not detected Target-Sequence 35S promotor
LLC “Spectrtrading” 1) Qualitative 35S/tNOS GMO Detection Kit;
1 step extraction with PrepMan ®ultra Sample Preparation Reagent proceid by Applied Biosystems by Life Technologies
Congen: Sure Food GMO/ Screen 35S/NOS/FMV
extraction with SureFood Prep Advaced
Biotecon Diagnostics
35S-Promoter DNA in FAM-Kanal SureFood® GMO Screen 4plex 35S/NOS/FMV+IAC
Extraction by SureFood® PREP DNA/RNA Virus
unknown Biotecon Biotecon foodproof Sample Prep. Kit III (S 400 06.1)Promotor des Cauliflower Mosaic
Virus (CaMV35S)Machery&Nagel NucleoSpin Food Kit / 200 mg sample weight
ASU § 64 LFGB L 15.05-1 (SDS/Guanidinium chloride-buffer with Proteinase K, clean-up by Wizard-Kit from Fa. Promega)
R-Biopharm R-Biopharm, SureFood PREP Advanced, according to manualTarget - DNA Biotecon Diagnostics foodproof GMO Sample Preparation Kit
Biotecon Biotecon DNA preapration kitfoodproof GMO Screening 1 LyoKit-5'Nuclease-, R 602 17-1, biotecon
DNA extraction according to foodproof Magnetic Preparation Kit III, biotecon S 400 13L
Macherey Nagel - nucleobondmodified CTAB-method with clean-up
Microsynth Pentaplex AllGVOscB
Macherey-Nagel Nucleospin Food Kit with own optimization; Proteinase K and RNase A; Clean-up via Silica columns; DNA-content and -quality photometric determination: sample 1: 48 ng/µl; A260/A280=1.8 A260/A230=2.1; sample 2: 48 ng/µl;
A260/A280=1,8 A260/A230=2,2 sample 3: 97ng/µl; A260/A280=1,9 A260/A230=1,5 sample 4: 146 ng/µl; A260/A280=1,9 A260/A230=1,6 sample 5: 67 ng/µl;
A260/A280=1,9 A260/A230=1,5
Congen 35S / nos / FMV Monoplex Congen PREP Basic extraction kit
August 2015 DLA – 24/2015 – GMO-Screening qualitative
5.1.2 NOS-Screening Sequence
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NOS
1
2
3 §64 LFGB
45
6
7
8
9
10 unbekannt
11 ASU L 00.00-122
12 ASU L 00.00-122
14 NOS1516 35 S, NOS
18 T-NOS
1920 ASU L00.00.31
21
22 GEN-IAL24
Evaluation number
Result given as Test-Kit or Literature Remarks to DNA-Extraction
Target-Sequenz / -DNA Supplier / Method e.g. Extraction / Enzymes / Clean-Up / DNA-Quality
extract. 2g with CTAB and clean-up with innuPREP DNA Micro Kit (845-KS-1010050; Analytik Jena)
QIAamp® DNA Stool Mini Kit (Fa. Qiagen)combination: CTAB-Lysis with Macherey Nagel NucleoSpin® Food
KitNOS-Terminator SureFood GMO Kit, R-Biopharm NucleoSpin Food, Fa. Machery & Nagel
Gen-ial FFS-Kit, Promegadetected or not detected Target-
Sequence NOS terminator2) 35S soybean qualitative/quantitative
GMO Detection Kit; Quality assessment of DNA extract was not conducted
Congen: Sure Food GMO/ Screen 35S/NOS/FMV
Biotecon Diagnostics
NOS-Terminator DNA in Cy5-KanalSureFood® GMO Screen 4plex
35S/NOS/FMV+IACExtraction by SureFood® PREP DNA/RNA Virus
Biotecon Biotecon foodproof Sample Prep. Kit III (S 400 06.1)Terminator des Agrobacterium
tumefaciens (nos)Machery&Nagel NucleoSpin Food Kit / 200 mg sample weight
tNOS-spezifische DNA-SequenzenASU § 64 LFGB L 15.05-1 (SDS/Guanidinium chloride-buffer with
Proteinase K, clean-up by Wizard-Kit from Fa. Promega)R-Biopharm R-Biopharm, SureFood PREP Advanced, according to manual
Target - DNA Biotecon Diagnostics foodproof GMO Sample Preparation KitBiotecon Biotecon DNA preapration kit
foodproof GMO Screening 1 LyoKit-5'Nuclease-, R 602 17-1, biotecon
DNA extraction according to foodproof Magnetic Preparation Kit III, biotecon S 400 13L
Macherey Nagel - nucleobondmodified CTAB-method with clean-up
Nos-terminator Microsynth Pentaplex AllGVOscBMacherey-Nagel Nucleospin Food Kit with own optimizations;
Proteinase K and RNase A; Clean-up with Silica columns;
nos Congen 35S / nos / FMV Monoplex Congen PREP Basic Extraction kit
August 2015 DLA – 24/2015 – GMO-Screening qualitative
5.1.3 GMO-Soya (RR-Round-Up-Ready-Soya)
5.1.4 Lectin-DNA (Soya-specific)
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1 L 00.00 1052
3 §64 LFGB
5
6
11
12
13 P35S/CTP1517
18
19
21 GTS 40-3-2
23
Evaluation number
Result given as Test-Kit or Literature Remarks to DNA-Extraction
RR-Soya Target-Sequenz / -DNA Supplier / Method e.g. Extraction / Enzymes / Clean-Up / DNA-Quality
see aboveQIAamp® DNA Stool Mini Kit (Fa. Qiagen)
combination: CTAB-Lysis with Macherey Nagel NucleoSpin® Food Kit
Gen-ial FFS-Kit, Promegadetected or not detected Soya RR 40-3-2 event-specific sequence
3) 35S maize qualitative/quantitative GMO Detection Kit;
Machery&Nagel NucleoSpin Food Kit / 200 mg sample weightp35S-CTP1 Konstrukt spezifische
DNA-SequenzenASU L 00.00-105, Annex C2
ASU § 64 LFGB L 15.05-1 (SDS/Guanidinium chloride-buffer with Proteinase K, clean-up by Wizard-Kit from Fa. Promega)
ASU L 00.00-105 Annex C.2 CTAB, Proteinase K, RNase A + QIAquickTarget - DNA Biotecon Diagnostics foodproof GMO Sample Preparation Kit
Target-Sequence EURL-Method Nucleo-Spin Food Kit (Maccherey & Nagel)
RRs foodproof GMO RR Soya Quantification
Kit-5'Nuclease-, R 302 19, bioteconDNA extraction according to foodproof Magnetic Preparation Kit III,
biotecon S 400 13L
Macherey Nagel - nucleobond
Microsynth Pentaplex All SoyMacherey-Nagel Nucleospin Food Kit with own optimizations;
Proteinase K and RNase A; Clean-up with Silica columns; Sure Food
Lectin
1 L 00.00 1052
3 §64 LFGB
5
6
11 Lectin-Gene ASU L 00.00-105
12
13 Lectin-Gene17
18
19
21 Lectin
Evaluation number
Result given as Test-Kit or Literature Remarks to DNA-Extraction
Target-Sequenz / -DNA Supplier / Method e.g. Extraction / Enzymes / Clean-Up / DNA-Quality
see aboveQIAamp® DNA Stool Mini Kit (Fa. Qiagen)
combination: CTAB-Lysis with Macherey Nagel NucleoSpin® Food Kit
Gen-ial FFS-Kit, Promegadetected or not detected taxon-specific sequence soy (fragment
lectin gen)
4) RR40-3-2 GMO line soybean qualitative Detection Kit;
Machery&Nagel NucleoSpin Food Kit / 200 mg sample weight
Lectin specific DNA-Sequences ASU L 00.00-105, Annex C2ASU § 64 LFGB L 15.05-1 (SDS/Guanidinium chloride-buffer with
Proteinase K, clean-up by Wizard-Kit from Fa. Promega)ASU L 00.00-105 Annex C.2 CTAB, Proteinase K, RNase A + QIAquick
Target-Sequence EURL-Methode Nucleo-Spin Food Kit (Maccherey & Nagel)
lectinfoodproof GMO RR Soya Quantification
Kit-5'Nuclease-, R 302 19, bioteconDNA extraction according to foodproof Magnetic Preparation Kit III,
biotecon S 400 13L
Macherey Nagel - nucleobond
Microsynth Pentaplex AllGVOscBMacherey-Nagel Nucleospin Food Kit with own optimizations; Proteinase K and RNase A; Clean-up with Silica columns;
August 2015 DLA – 24/2015 – GMO-Screening qualitative
5.1.5 GMO-Maize (bt11-Maize)
5.1.6 Maize-DNA (Maize-specific)
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bt11-Maize
1
3 §64 LFGB
5
6
13 ASU L 15.05-11517
18 Bt11
19
21 Bt-11
Evaluation number
Result given as Test-Kit or Literature Remarks to DNA-Extraction
Target-Sequenz / -DNA Supplier / Method e.g. Extraction / Enzymes / Clean-Up / DNA-Quality
see abovecombination: CTAB-Lysis with Macherey Nagel NucleoSpin® Food
KitGen-ial FFS-Kit, Promega
detected or not detected Maize bt11 event-specific sequence
5) bt11 GMO line maize qualitative Detection Kit
IVS2-2/PAT-B-Primer CTAB, Proteinase K, RNase A + QIAquickTarget - DNA Biotecon Diagnostics foodproof GMO Sample Preparation Kit
Target-Sequence EURL-Method Nucleo-Spin Food Kit (Maccherey & Nagel)SureFood GMO QUANT Bt11 Corn,
S2016, r-biopharmDNA extraction according to foodproof Magnetic Preparation Kit III,
biotecon S 400 13LMacherey Nagel - nucleobond
JRC protocol event-specific method for the quantification of maize event Bt11
Macherey-Nagel Nucleospin Food Kit with own optimizations; Proteinase K and RNase A; Clean-up with Silica columns;
1
3 §64 LFGB
5
6
11 ASU L 00.00-105
12
131517
18
19
21
Evaluation number
Result given as Test-Kit or Literature Remarks to DNA-Extraction
Target-Sequenz / -DNA Supplier / Method e.g. Extraction / Enzymes / Clean-Up / DNA-Quality
see abovecombination: CTAB-Lysis with Macherey Nagel NucleoSpin® Food
KitGen-ial FFS-Kit, Promega
detected or not detected taxon-specific sequence maize (fragment
zein gen)
Made from reagents manufactured by Applied Biosystems/ Life Technologies
hmg-Gen Machery&Nagel NucleoSpin Food Kit / 200 mg sample weight
Invertase specific DNA-SequencesBrodmann et al. (2002) J. of AOAC Int.
85(3): 646-653ASU § 64 LFGB L 15.05-1 (SDS/Guanidinium chloride-buffer with
Proteinase K, clean-up by Wizard-Kit from Fa. Promega)HMG-Gene ASU L 00.00-105 Anh. D.2 CTAB, Proteinase K, RNase A + QIAquick
Target - DNA Biotecon Diagnostics foodproof GMO Sample Preparation KitTarget-Sequence EURL-Method Nucleo-Spin Food Kit (Maccherey & Nagel)
zeinSureFood GMO QUANT Bt11 Corn,
S2016, r-biopharmDNA extraction according to foodproof Magnetic Preparation Kit III,
biotecon S 400 13L
Macherey Nagel - nucleobond
mhmg Microsynth Pentaplex AllGVOscBMacherey-Nagel Nucleospin Food Kit with own optimizations; Proteinase K and RNase A; Clean-up with Silica columns;
August 2015 DLA – 24/2015 – GMO-Screening qualitative
5.1.7 Other Parameters (DNA)
Reprint, also in part, only with written permission from DLA-AhrensburgPage 17 of 25
1a
1b
1c
1d
1e24 Promotor578
11 ASU L 00.00-125
12a ASU G 30.40-1
12b
12c ASU L 00.00-125
12d
12e ASU L 00.00-141
14 FMV15
20
21a
21b CP2 & CP4 EPSPS
21c Bar-Gen
21d
21e MON89788
24 FMV
Evaluation number
Result given as Test-Kit or Literature Remarks to DNA-Extraction
Target-Sequenz / -DNA Supplier / Method e.g. Extraction / Enzymes / Clean-Up / DNA-Quality
see above [tables]
see above [tables]
see above [tables]
see above [tables]
see above [tables]QIAamp® DNA Stool Mini Kit (Fa. Qiagen)
SureFood GMO Kit, R-Biopharm NucleoSpin Food, Fa. Machery & NagelGen-ial FFS-Kit, Promega
Biotecon DiagnosticsTransition from CTP2 to CP4-
EPSPS-GeneMachery&Nagel NucleoSpin Food Kit / 200 mg sample weight
p35S-pat construct specific DNA-Sequence
ASU § 64 LFGB L 15.05-1 (SDS/Guanidinium chloride-buffer with Proteinase K, clean-up by Wizard-Kit from Fa. Promega)
p35S-pat construct specific DNA-sequences
Methodensammlung der Länderarbeitsgemeinschaft Gentechnik
(AM001)
ASU § 64 LFGB L 15.05-1 (SDS/Guanidinium chloride-buffer with Proteinase K, clean-up by Wizard-Kit from Fa. Promega)
CTP2-CP4-EPSPS construct specific DNA-sequences
ASU § 64 LFGB L 15.05-1 (SDS/Guanidinium chloride-buffer with Proteinase K, clean-up by Wizard-Kit from Fa. Promega)
bar-specific DNA-sequences ASU L 00.00-124, modifiedASU § 64 LFGB L 15.05-1 (SDS/Guanidinium chloride-buffer with
Proteinase K, clean-up by Wizard-Kit from Fa. Promega)
pNOS specific DNA-sequencesASU § 64 LFGB L 15.05-1 (SDS/Guanidinium chloride-buffer with
Proteinase K, clean-up by Wizard-Kit from Fa. Promega)R-Biopharm R-Biopharm, SureFood PREP Advanced, nach Anleitung
Target - DNA Biotecon Diagnostics foodproof GMO Sample Preparation KitScreening Plant-DNA / in-house-
methodmodifiziertes CTAB-Verfahren mit clean-up
Pat-Gen Microsynth Tetraplex AllGVOscCMacherey-Nagel Nucleospin Food Kit with own optimizations;
Proteinase K and RNase A; Clean-up with Silica columns;
Microsynth Tetraplex AllGVOscCMacherey-Nagel Nucleospin Food Kit with own optimizations;
Proteinase K and RNase A; Clean-up with Silica columns;
Microsynth Tetraplex AllGVOscCMacherey-Nagel Nucleospin Food Kit with own optimizations;
Proteinase K and RNase A; Clean-up with Silica columns;
FMV promoter Microsynth Tetraplex AllGVOscCMacherey-Nagel Nucleospin Food Kit with own optimizations;
Proteinase K and RNase A; Clean-up with Silica columns;
Microsynth Pentaplex All SoyMacherey-Nagel Nucleospin Food Kit with own optimizations;
Proteinase K and RNase A; Clean-up with Silica columns; Congen 35S / nos / FMV Monoplex Congen PREP Basic Extraction kit
August 2015 DLA – 24/2015 – GMO-Screening qualitative
5.2 Details by participants to PCR-reaction
5.2.1 35S-Screening Sequence
Reprint, also in part, only with written permission from DLA-AhrensburgPage 18 of 25
1 Real Time PCR2
3 Real Time PCR 4
5 Real Time PCR
6
7
8
9
10
11
12
14
15
16 Real time PCR18
19 real time PCR20
21
22
24
Evaluation number
Notes to PCR-Reaction Further Remarks
e.g. Real Time PCR / Gel electrophoresis / Cycles / Lenght of Amplif icates / Refe-rence material
Gel electrophoresis/ 45 Cycles/ 123 bp/ Bt11-Maize, RR-Soya tested in double determination
RT PCR, 45 Cycles
Real time PCR ( 7300 Real time PCR system). PCR protocol (amplification) : 1) stage 1 (initial steps: AmpEraseUNG activation) 2 min 50 °C - 1 cycles; 2) stage 2 (initial steps: Ampli Tag DNA polymerase
activation) 10 min 95 °C - 1 cycles; 3) stage 3 ( each of 40 cycles) denaturation 15 sec 95 °C, anneal/extend 1 min 58 °C
Sample 2: 35S in the range of LODReal Time PCR /Multiplex
Real Time PCR, 45 Cycles, internal Amplification controlreal-time PCR (Biotecon foodproof GMO Screening Kit (R 302 17))
Real-time PCR; 45 Cycles; 82 bp; Reference material gv-Maize Bt11Sample 3: traces below absolute LOD
(valuated negative)Real Time PCR, SureFood GVO 4plex, R-Biopharm, according to
manualfoodproof GMO Screenig Kit
real time PCR/ 50 cycles
Gel electrophoresisReal-Time PCR m. 45 Cycles; certified reference material: GTS 40-3-2 Soya (IRMM) with 0.1%, 1% and 10% GMO-content; for control of
inhibition sample eluates were tested in 1:5 and 1:25 dilutions; negative control (Nuclease-free H2O instead of DNA)
Real Time PCR, 45 Cycles, Reference material ERM-BF 410dk * Sample 3: traces ct 38,5
August 2015 DLA – 24/2015 – GMO-Screening qualitative
5.2.2 NOS-Screening Sequence
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1 Real Time PCR2
3 Real Time PCR 4
5 Real Time PCR
6
7
8
9
10
11
12
14
15
16 Real time PCR18
19 real time PCR20
21
22
24
Evaluation number
Notes to PCR-Reaction Further Remarks
e.g. Real Time PCR / Gel electrophoresis / Cycles / Lenght of Amplif icates / Refe-rence material
Gel electrophoresis/ 45 Cycles/ 123 bp/ Bt11-Maize, RR-Soya tested in double determination
RT PCR, 45 Cycles
PCR controls: 1) PCR inhibition control - reaction mix with plasmid DNA ; 2) PCR reagent control (negative control) - reaction mix with nucleic acid free water; 3) PCR positive DNA target control (positive control) - reaction mix with DNA extact (10% RR 40-3-2 soya; 10 % bt11; 2% MON 810 test-
kit №3)
Real Time PCR /Multiplex
Real Time PCR, 45 Cycles, internal Amplification controlreal-time PCR (Biotecon foodproof GMO Screening Kit (R 302 17))
Real-time PCR; 45 Cycles; 84 bp; Reference material gv-Maize Bt11Sample 3: traces below absolute LOD
(valuated negative)Real Time PCR, SureFood GVO 4plex, R-Biopharm, according to
manualfoodproof GMO Screenig Kit
real time PCR/ 50 cycles
Gel electrophoresisReal-Time PCR m. 45 Cycles; certified reference material: GTS 40-3-2 Soya (IRMM) with 0.1%, 1% and 10% GMO-content; for control of
inhibition sample eluates were tested in 1:5 and 1:25 dilutions; negative control (Nuclease-free H2O instead of DNA)
Real Time PCR, 45 Cycles, Reference material ERM-BF 410dk * Sample 3: traces ct 38,8
August 2015 DLA – 24/2015 – GMO-Screening qualitative
5.2.3 GMO-Soya (RR-Round-Up-Ready-Soya)
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1 Real Time PCR
23 Real Time PCR
5 Real Time PCR
6
11
12
1315
17
1819 real time PCR
21
23 real time PCR
Evaluation number
Notes to PCR-Reaction Further Remarks
e.g. Real Time PCR / Gel electrophoresis / Cycles / Lenght of Amplif icates / Refe-rence material
Gel electrophoresis/ 45 Cycles/ 169 bp/ RR-Soya tested in double determination
Сertified reference material: Institute of Reference Materials and Measurements,Geel Belgium
Real-time PCR; 45 Cycles; 74 bp; Reference material gm-Soya GTS 40-3-2
Sample 3: traces below relative LOD (valuated negative)
Real-time PCR 74 bp
foodproof GMO Screenig Kit
Real Time PCR / 45 Cycles / Reference material JRC (Geel Belgium) /10 to 100 ng DNA per well
real time PCR/ 50 cycles
Real-Time PCR m. 45 Cycles; certified reference material: GTS 40-3-2 Soya (IRMM) with 0.1%, 1% and 10% GMO-content, as well as 0%
GMO-Soya. For identification 200 ng (40 ng/µl) DNA each was applied. Negative control (Nuclease-free H2O instead of DNA)
Sample 4: RR-content in total Soya <0,1%; Sample 5: RR-content 5,2%
August 2015 DLA – 24/2015 – GMO-Screening qualitative
5.2.4 Lectin-DNA (Soya-specific)
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1 Real Time PCR
23 Real Time PCR
5 Real Time PCR
6
11
12
13
17
1819 real time PCR
21
Evaluation number
Notes to PCR-Reaction Further Remarks
e.g. Real Time PCR / Gel electrophoresis / Cycles / Lenght of Amplif icates / Refe-rence material
Gel electrophoresis/ 45 Cycles/ 438 bp/ RR-Soya tested in double determination
Roundup ReadyTM soya beans (ERM-BF410 ak; ERM-BF410 bk; ERM-BF410 dk; ERM-BF410 gk)
Real-time PCR; 45 Cycles; 74 bp; Reference material gm-Soya GTS 40-3-2
Real-time PCR 74 bpSamples 1 + 2 showed late amplification
curves, not clearly positiveReal Time PCR / 45 Cycles / Reference material JRC (Geel Belgium) /10
to 100 ng DNA per well
real time PCR/ 50 cycles
Real-Time PCR m. 45 Cycles; certified reference material: GTS 40-3-2 Soya (IRMM) with 0.1%, 1% and 10% GMO-content; for control of
inhibition sample eluates were tested in 1:5 and 1:25 dilutions; negative control (Nuclease-free H2O instead of DNA)
Sample 1 low soya traces
August 2015 DLA – 24/2015 – GMO-Screening qualitative
5.2.5 GMO-Maize (bt11-Maize)
5.2.6 Maize-DNA (Maize-specific)
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1 Real Time PCR
3 Real Time PCR
5 Real Time PCR
6
1315
17
1819 real time PCR
21
Evaluation number
Notes to PCR-Reaction Further Remarks
e.g. Real Time PCR / Gel electrophoresis / Cycles / Lenght of Amplif icates / Refe-rence material
MON 810 maize (ERM-BF413 ak; ERM-BF413 ck; ERM-BF413 ek; ERM-BF413 gk;)
PCR 189 bp + Gel electrophoresis + Restriction 116+73 bp
foodproof GMO Screenig Kit
Real Time PCR / 45 cycles / Reference material JRC (Geel Belgium) /10 to 100 ng DNA per well
real time PCR/ 50 cycles
Real-Time PCR 45 cycles; certified reference material: Bt-11 Maize (IRMM) with 0.1%, 1% and 5% GMO-content; For identification 200 ng (40
ng/µl) DNA each were applied. Negative control (Nuclease-free H2O instead of DNA)
Sample 2: 0,9% Bt-11 in total Maize
1 Real Time PCR
3 Real Time PCR
5 Real Time PCR
6
11
12
1315
17
1819 real time PCR
21
Evaluation number
Notes to PCR-Reaction Further Remarks
e.g. Real Time PCR / Gel electrophoresis / Cycles / Lenght of Amplif icates / Refe-rence material
Bt-11 maize (ERM-BF412 f)
Samples 3 and 5 not suitable for analysis of gm-maize, they contain too less amplifyable maize DNA. Sample 4: sample specific LOD
of gm-maize 0,7 %.
Real-time PCR; 45 cycles; 79 bp; Reference material gm-maize Bt11
Samples 3 and 5: traces of maize (valuation negative);
sample 4: clear traces of maize (contamination?) detected (valuation
positive)
Real-time PCR, 79 bp
foodproof GMO Screenig Kit
Real Time PCR / 45 cycles / Reference material JRC (Geel Belgium) /10 to 100 ng DNA per well
real time PCR/ 50 cycles
Real-Time PCR 45 cycles; for inhibition control sample eluates were 1:5 and 1:25 diluted
Samplles 3+4: low maize traces
August 2015 DLA – 24/2015 – GMO-Screening qualitative
5.2.7 Other Parameters (DNA)
Reprint, also in part, only with written permission from DLA-AhrensburgPage 23 of 25
1a Real Time PCR
1b Real Time PCR
1c Real Time PCR
1d Real Time PCR
1e Real Time PCR
245 Real Time PCR
78 bar-gen,/FMV
11
12a
12b
12c
12d12e141520
21a
21b
21c
21d
21e
24
Evaluation number
Notes to PCR-Reaction Further Remarks
e.g. Real Time PCR / Gel electrophoresis / Cycles / Lenght of Amplif icates / Refe-rence material
multiple extraction dif fering results, at LOD?
Gel electrophoresis/ 45 Cycles/ 200-600 bp/ Bt11-Maize, Bt176, RR-Soya tested in double determination
RT PCR, 45 Cycles
Real Time PCR /Multiplex
Real-time PCR; 45 cycles; 102/111 and 402 bp (Bt11) respectively; Reference material gm-Maize Bt11
conventional PCR-RFLP; 45 cycles; 370 and 550 bp (Bt11); Reference material gm-Maize Bt11 and T25
Sample 2: size of PCR-amplif icates and restriction fragments of positive sample 2 not in agreement
w ith gm-maize Bt11
Real-time PCR; 45 cycles; 88 bp; Reference material gm-rape Gt73Samples 4 and 5: clear traces (contamination?)
detected (valuated positive)
Real-time PCR; 45 cycles; 110 bp; reference material gm-rape Ms1xRf1
Real-time PCR; 45 cycles; 94 bp; reference material gm-rape Ms1xRf1
Real Time PCR, SureFood GVO 4plex, R-Biopharm, according to manual
foodproof GMO Screenig Kit
Gel electrophoresis
Real-Time PCR 45 cycles; certif ied reference material: MON89788 Soya (IRMM) w ith 0.1%, 1% and 10% GMO-content. negative control (Nuclease-free H2O
instead of DNA)
Real-Time PCR 45 cycles; certif ied reference material: MON89788 Soya (IRMM) w ith 0.1%, 1% and 10% GMO-content. negative control (Nuclease-free H2O
instead of DNA)
Real-Time PCR 45 cycles; negative control (Nuclease-free H2O instead of DNA)
Real-Time PCR 45 cycles; negative control (Nuclease-free H2O instead of DNA)
Real-Time PCR 45 cycles; certif ied reference material: MON89788 Soya (IRMM) w ith 0.1%, 1% and 10% GMO-content, as w ell as 0% GMO-Soya. For identif ication 200 ng (40 ng/µl) DNA w ere applied. Negative control (Nuclease-f ree H2O instead
of DNA)
Sample 5: MON89788-content in total soya <0,1%
Real Time PCR, 45 Cycles, reference material ERM-BF 410dk * Sample 4: traces ct 38,2
August 2015 DLA – 24/2015 – GMO-Screening qualitative
6. Index of participant laboratories
[Die Adressdaten der Teilnehmer wurden für die allgemeine Veröffentli-chung des Auswerte-Berichts nicht angegeben.]
[The address data of the participants were deleted for publication of the evaluation report.]
Reprint, also in part, only with written permission from DLA-AhrensburgPage 24 of 25
FRANCE
AUSTRIABELARUSNETHERLANDS
Teilnehmer / Participants Ort / TownGermanyGermanyGermany
GermanyGermanyGermanyGermanyGermanyGermanyGermanyGermanyGermanyGermanyGermanyGermanyGermanyGermany
GermanyGermany
August 2015 DLA – 24/2015 – GMO-Screening qualitative
7. Index of references
1. DIN EN ISO/IEC 17043:2010; Konformitätsbewertung – Allgemeine Anforderungen an Eignungsprüfungen / Conformity assessment – General requirements for proficiency testing
2. Verordnung / Regulation 882/2004/EU; Verordnung über amtliche Kontrollen / Regulation on official controls
3. DIN EN ISO/IEC 17025:2005; Allgemeine Anforderungen an die Kompetenz von Prüf- und Kalibrierlaboratorien / General requirements for the competence of testing and calibration laboratories
4. Richtlinie / Directive 1993/99/EU; über zusätzliche Maßnahmen im Bereich der amtlichen Lebensmittelüberwachung / on additional measuresconcerning the official control of foodstuffs
5. ASU §64 LFGB : Planung und statistische Auswertung von Ringversuchen zur Methodenvalidierung
6. DIN ISO 13528:2009; Statistische Verfahren für Eignungsprüfungen durchRingversuche / Statistical methods for use in proficiency testing by interlaboratory comparisons
7. The International Harmonised Protocol for the Proficiency Testing of Ananlytical Laboratories ; J.AOAC Int., 76(4), 926 – 940 (1993)
8. The International Harmonised Protocol for the Proficiency Testing of Ananlytical Chemistry Laboratories ; Pure Appl Chem, 78, 145 – 196 (2006)
9. Evaluation of analytical methods used for regulation of food and drugs;W. Horwitz; Analytical Chemistry, 54, 67-76 (1982)
10.A Horwitz-like funktion describes precision in proficiency test; M. Thompson, P.J. Lowthian; Analyst, 120, 271-272 (1995)
11.Protocol for the design, conduct and interpretation of method performance studies; W. Horwitz; Pure & Applied Chemistry, 67, 331-343(1995)
12.Recent trends in inter-laboratory precision at ppb and sub-ppb concentrations in relation to fitness for purpose criteria in proficiency testing; M. Thompson; Analyst, 125, 385-386 (2000)
13.European Network of GMO Laboratories, Definition of Minimum Performance Requirements for Analytical Methods of GMO Testing, Version 25-01-2005
14.Powell J, Owen L, Reliability of food measurements: the application of proficiency testing to GMO analysis, Accred Qual. Assur. 7, 392-402(2002)
15.Thompson M, GMO Proficiency testing: Interpreting z-scores derived from log-transformed data, amc technical brief, No. 18 Dec 2004
16.Thompson M et al., Scoring in Genetically Modified Organism Proficiency Tests Based on Log-Transformed Results, J. AOAC Int., 89(1), 232-239 (2006)
17.Žel J et al., Calculation of Measurement Uncertainty in Quantitative Analysis of Genetically Modified Organisms Using Intermediate Precision - A Practical Approach, J. AOAC Int., 90(2), 582-586 (2007)
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