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Quantitative Testing Plans May 8-10, 2006 Iowa State University, Ames – USA Jean-Louis Laffont Kirk Remund Overview • Statistical framework • Implementation in Seedcalc ISTA Statistics Committee 2 Testing plan design: statistical framework for quantitative methods • Suppose n pools of m seeds were taken from the lot and that J flour sub-samples from each pool were measured K times. Seed lot: true AP% = p Grinding seeds into flour … … J flour sub-samples per pool Measurement n pools of m seeds … … yijk K measures per flour sub-sample Measure 1 Measure 2 Pool i … Measure K Flour sub-sam ple 1 Flour sub-sam ple 2 … Flour sub-sam ple J ISTA Statistics Committee 3 Testing plan design: statistical framework for quantitative methods • Model: y ijk p a i b j(i ) e ijk Measurement k made on flour sub-sample j from pool i = True AP% + Random effect of pool i 2 N 0, sampling + Random effect of flour sub-sample j from pool i 2 N 0, flour + Random effect of measurmnt k for flour sub-sample j from pool i N 0, 2measuremen t The parameter p is estimated by the sample mean: 1 2 p̂ y ~ N p, ijk p̂ nJK i , j,k 2 sampling 2 flour 2 measuremen t n nJ nJK 2 p̂ ISTA Statistics Committee 4 Testing plan design: statistical framework for quantitative methods Overall distribution of y ~ N(p, 2 y ) yijk 2 sampling 2 flour 2 measuremen t n nJ nJK 2 p̂ 0.0 0.5 1.0 1.5 2.0 Sampling n pools of m seeds AP% 2 : derived from the sampling variance of B(m, p ker nel ) p + 2 sampling p ker nel (1 p ker nel ) m Flour sub-sampling -1.0 -0.5 0.0 0.5 1.0 2 flour AP% 2 flour and 2 measuremen t are obtained from historical experiments Measurement 2measuremen t ISTA Statistics Committee 5 Testing plan design: statistical framework for quantitative methods • Remember that the true AP% p in the lot is expressed in %DNA when using quantitative methods p ker nel (1 p ker nel ) 2 and that sampling is expressed on a m kernel basis. Introduction of the b-Factor (biological factor) to convert from %DNA to %Seed units: %Seed = b-Factor x %DNA or pker nel b p • Examples: • Reference material and test lots have the same zygosity/ploidy/copy number b-Factor= 1 • Homozygous reference material and hemizygous test lots b-Factor= 2 ISTA Statistics Committee 6 Testing plan design: statistical framework for quantitative methods • Re-expression of 2 2 • Re-expression of 2 sampling sampling p̂ in %DNA units: p(1 bp) bm 2 sampling 2 flour 2 measuremen t n nJ nJK : Having observed in some experiments that ²measurement seems to depend on p, the true AP probability, while CVmeasurement is fairly constant, we can rewrite 2 p̂ as: 2 p̂ p(1 bp) 2flour (pCVmeasurement )2 bnm nJ nJK ISTA Statistics Committee 7 Testing plan design: statistical framework for quantitative methods • Lets now define an Acceptance Limit (AL) such that: • if p̂ AL, “accept” the lot • if p̂ > AL, “reject” the lot • We can then calculate the probability to “accept” the lot, given a true unknown AP% p: p̂ p AL p AL p where F is the cumulative distribution Pr( p̂ AL | p) Pr | p F function for the standard normal distribution p̂ p̂ p̂ • This formula serves as a basis for elaborating an OC curve that can be used to investigate properties of a testing plan ISTA Statistics Committee 8 Testing plan design: statistical framework for quantitative methods • Example: testing plan components: 2 pools of 3000 seeds, 100 1 flour sub-sample/pool, 3 measurements/flour sub-sample, Std-Dev of flour sub-sampling error = 0.011%, measurement CV = 15%, Acceptance Limit (AL) = 0.1% (lot « accepted » if average of the 2 x 1 x 3 readings is AL) 60 40 20 5% 0 Probability of acceptance (%) 80 95% 0.0 0.1 0.2 0.3 0.4 0.5 True AP% in lot ISTA Statistics Committee 9 Testing plan design: statistical framework for quantitative methods • Consumer risk and producer risk are given respectively by: AL LQL Consumer risk Pr( p̂ AL | LQL) F p̂ AL AQL Pr oducer risk Pr( p̂ AL | AQL) 1 F p̂ where F is the cumulative distribution function for the standard normal distribution ISTA Statistics Committee 10 Testing plan design: implementation for quantitative methods • All of the methods discussed have been implemented in the newest version of the Microsoft Excel® spreadsheet Seedcalc Estimating AP% Designing testing plans ISTA Statistics Committee Comparing testing plans 11 Testing plan design: implementation for quantitative methods Testing plan design LQL, AQL and AL Enter n, m, J, K and … historical assay variation and… b-Factor and … and get consumer and producer risks and OC curve ISTA Statistics Committee 12 Testing plan design: implementation for quantitative methods The « Find Plan » tool can help the user to find testing plans satisfying certain conditions given some parameters ISTA Statistics Committee 13 Testing plan design: implementation for quantitative methods Parameters for the search algorithms ISTA Statistics Committee 14 Testing plan design: implementation for quantitative methods Find the highest AL that meets target consumer risk for the LQL. No consideration of the producer risk target. n, m, J and K are held fixed ISTA Statistics Committee 15 Testing plan design: implementation for quantitative methods Consumer and producer risk targets satisfied by changing AL, n, I and J ISTA Statistics Committee 16 Testing plan design: implementation for quantitative methods Consumer and producer risk targets satisfied by changing AL, m, I and J ISTA Statistics Committee 17 Testing plan design: implementation for quantitative methods Consumer and producer risk targets satisfied by changing AL, n, m, I and J ISTA Statistics Committee 18 Testing plan design: implementation for quantitative methods Compare plans Visual comparison of OC curves along with testing plan parameters ISTA Statistics Committee 19 Example Historical data gave 0.15% for the estimate of the flour standard-deviation. We expect that the measurement CV range is from 10% to 30% and we consider the following testing plan: . LQL = 0.7% for a consumer confidence = 95% . AQL = 0.15% for a producer confidence = 95% . 1 pool of 3000 seeds, 2 flour sub-samples, 3 measurements . AL = 0.39% 1. Does this plan meet consumer and producer requirements when the measurement CV = 10%? 2. Compare the outcomes of this testing plan when the CV is varying from 10% to 30%. ISTA Statistics Committee 20 Example 1. Does this plan meet consumer and producer requirements when the measurement CV = 10%? YES ISTA Statistics Committee 21 Example 2. Compare the outcomes of this testing plan when the CV is varying from 10% to 30%. ISTA Statistics Committee 22