Performance standard-based validation study for local lymph node assay: 5-bromo-2-deoxyuridine-flow cytometry method

Ilyoung Ahn, Tae Sung Kim, Eun Sun Jung, Jung Sun Yi, Won Hee Jang, Kyoung Mi Jung, Miyoung Park, Mi Sook Jung, Eun Young Jeon, Kyeong uk Yeo, Ji Hoon Jo, Jung Eun Park, Chang Yul Kim, Yeong Chul Park, Won Keun Seong, Ai Young Lee, Young Jin Chun, Tae Cheon Jeong, Eui Bae Jeung, Kyung Min LimSeung Jin Bae, Soojung Sohn, Yong Heo

Research output: Contribution to journalArticlepeer-review

12 Scopus citations

Abstract

Local lymph node assay: 5-bromo-2-deoxyuridine-flow cytometry method (LLNA: BrdU-FCM) is a modified non-radioisotopic technique with the additional advantages of accommodating multiple endpoints with the introduction of FCM, and refinement and reduction of animal use by using a sophisticated prescreening scheme. Reliability and accuracy of the LLNA: BrdU-FCM was determined according to OECD Test Guideline (TG) No. 429 (Skin Sensitization: Local Lymph Node Assay) performance standards (PS), with the participation of four laboratories. Transferability was demonstrated through successfully producing stimulation index (SI) values for 25% hexyl cinnamic aldehyde (HCA) consistently greater than 3, a predetermined threshold, by all participating laboratories. Within- and between-laboratory reproducibility was shown using HCA and 2,4-dinitrochlorobenzene, in which EC2.7 values (the estimated concentrations eliciting an SI of 2.7, the threshold for LLNA: BrdU-FCM) fell consistently within the acceptance ranges, 0.025–0.1% and 5–20%, respectively. Predictive capacity was tested using the final protocol version 1.3 for the 18 reference chemicals listed in OECD TG 429, of which results showed 84.6% sensitivity, 100% specificity, and 88.9% accuracy compared with the original LLNA. The data presented are considered to meet the performance criteria for the PS, and its predictive capacity was also sufficiently validated.

Original languageEnglish
Pages (from-to)183-194
Number of pages12
JournalRegulatory Toxicology and Pharmacology
Volume80
DOIs
StatePublished - Oct 2016

Bibliographical note

Publisher Copyright:
© 2016 Elsevier Inc.

Keywords

  • Flow cytometry
  • LLNA: BrdU-FCM
  • Local lymph node assay
  • Performance standards
  • Skin sensitization
  • Validation study

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