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Appraisal of within- and between-laboratory reproducibility of non-radioisotopic local lymph node assay using flow cytometry, LLNA: BrdU-FCM: Comparison of OECD TG429 performance standard and statistical evaluation

Title
Appraisal of within- and between-laboratory reproducibility of non-radioisotopic local lymph node assay using flow cytometry, LLNA: BrdU-FCM: Comparison of OECD TG429 performance standard and statistical evaluation
Authors
Yang, HyeriNa, JihyeJang, Won-HeeJung, Mi-SookJeon, Jun-YoungHeo, YongYeo, Kyung-WookJo, Ji-HoonLim, Kyung-MinBae, SeungJin
Ewha Authors
배승진임경민
SCOPUS Author ID
배승진scopus; 임경민scopus
Issue Date
2015
Journal Title
TOXICOLOGY LETTERS
ISSN
0378-4274JCR Link1879-3169JCR Link
Citation
vol. 234, no. 3, pp. 172 - 179
Keywords
Local lymph node assayLLNA:BrdU-FCMWithin and between laboratory reproducibilityOECD TG performance standardDescriptive and inferential statistics
Publisher
ELSEVIER IRELAND LTD
Indexed
SCI; SCIE; SCOPUS WOS scopus
Abstract
Mouse local lymph node assay (LLNA, OECD TG429) is an alternative test replacing conventional guinea pig tests (OECD TG406) for the skin sensitization test but the use of a radioisotopic agent, H-3-thymidine, deters its active dissemination. New non-radioisotopic LLNA, LLNA:BrdU-FCM employs a nonradioisotopic analog, 5-bromo-2'-deoxyuridine (BrdU) and flow cytometry. For an analogous method, OECD TG429 performance standard (PS) advises that two reference compounds be tested repeatedly and ECt(threshold) values obtained must fall within acceptable ranges to prove within- and between-laboratory reproducibility. However, this criteria is somewhat arbitrary and sample size of ECt is less than 5, raising concerns about insufficient reliability. Here, we explored various statistical methods to evaluate the reproducibility of LLNA: BrdU-FCM with stimulation index (SI), the raw data for ECt calculation, produced from 3 laboratories. Descriptive statistics along with graphical representation of SI was presented. For inferential statistics, parametric and non-parametric methods were applied to test the reproducibility of SI of a concurrent positive control and the robustness of results were investigated. Descriptive statistics and graphical representation of SI alone could illustrate the within- and between-laboratory reproducibility. Inferential statistics employing parametric and nonparametric methods drew similar conclusion. While all labs passed within-and between-laboratory reproducibility criteria given by OECD TG429 PS based on ECt values, statistical evaluation based on SI values showed that only two labs succeeded in achieving within-laboratory reproducibility. For those two labs that satisfied the within-lab reproducibility, between-laboratory reproducibility could be also attained based on inferential as well as descriptive statistics. (C) 2015 Elsevier Ireland Ltd. All rights reserved.
DOI
10.1016/j.toxlet.2015.02.014
Appears in Collections:
약학대학 > 약학과 > Journal papers
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