Seo Jung-Ah, Cho Sun-A, Park Chang Eon, Seo Dong Hyuk, Choi Myungsuk, An Susun, Kim Bae-Hwan
Department of Public Health, Keimyung University, 1095 Dalgubeol-daero, Dalseo-gu, Daegu, 42601 Republic of Korea.
Safety and Microbiology Lab, Amorepacific Corporation R&D Center, Yongin-si, Republic of Korea.
Toxicol Res. 2022 Jun 4;38(4):531-544. doi: 10.1007/s43188-022-00130-8. eCollection 2022 Oct.
Skin sensitization is induced when certain chemicals bind to skin proteins. Direct peptide reactivity assay (DPRA) has been adopted by the OECD as an alternative method to evaluate skin sensitization by assessing a substance's reaction to two model peptides. A modified spectrophotometric method, Spectro-DPRA, can evaluate skin sensitization, in a high throughput fashion, to obviate some limitations of DPRA. Pre-validation studies for Spectro-DPRA were conducted to determine transferability and proficiency, within- and between-laboratory reproducibility, and predictive ability based on GLP principles at three laboratories (AP, KTR, and KCL). All laboratories confirmed high (> 90%) concordance for evaluating the sensitivity induced by ten chemical substances. The concordance among the three tests performed by each laboratory was 90% for AP, 100% for KTR, and 100% for KCL. The mean accuracy of the laboratories was 93.3% [compared to the standard operating procedure (SOP)]. The reproducibility among the three laboratories was as high as 86.7%; the accuracy was 86.7% for AP, 100% for KTR, and 86.7% for KCL (compared to the SOP). An additional 54 substances were assessed in 3 separate labs to verify the prediction rate. Based on the result, 29 out of 33 substances were classified as sensitizers, and 19 out of 21 identified as non-sensitizers; the corresponding sensitivity, specificity, and accuracy values were 87.9%, 90.5%, and 88.9%, respectively. These findings indicate that the Spectro-DPRA can address the molecular initiating event with improved predictability and reproducibility, while saving time and cost compared to DPRA or ADRA.
当某些化学物质与皮肤蛋白质结合时，会引发皮肤致敏。经济合作与发展组织（OECD）采用直接肽反应性测定法（DPRA）作为一种替代方法，通过评估一种物质对两种模型肽的反应来评估皮肤致敏性。一种改进的分光光度法，即光谱-DPRA，可以高通量方式评估皮肤致敏性，以消除DPRA的一些局限性。基于GLP原则，在三个实验室（AP、KTR和KCL）进行了光谱-DPRA的预验证研究，以确定其转移性和熟练度、实验室内和实验室间的重现性以及预测能力。所有实验室在评估十种化学物质诱导的敏感性方面均确认了高一致性（>90%）。每个实验室进行的三项测试之间的一致性，AP为90%，KTR为100%，KCL为100%。实验室的平均准确率为93.3%[与标准操作程序（SOP）相比]。三个实验室之间的重现性高达86.7%；与SOP相比，AP的准确率为86.7%，KTR为100%，KCL为86.7%。在3个独立实验室中对另外54种物质进行了评估，以验证预测率。根据结果，33种物质中有29种被归类为致敏剂，21种已识别物质中有19种被确定为非致敏剂；相应的敏感性、特异性和准确率值分别为87.9%、90.5%和88.9%。这些发现表明，光谱-DPRA可以解决分子引发事件，具有更高的可预测性和重现性，同时与DPRA或ADRA相比节省了时间和成本。
