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Title: Manual Scoring with Shortened 48 h Cytokinesis-Block Micronucleus Assay Feasible for Triage in the Event of a Mass-Casualty Radiation Accident
Authors: Valerie Swee Ting Goh 
Yohei Fujishima
Ryo Nakayama
Kai Takebayashi
Mitsuaki A. Yoshida
Kosuke Kasai
Kentaro Ariyoshi
Tomisato Miura
Keywords: cytokinesis-block micronucleus assay
radiation accident
cytogenetic biodosimetry
Issue Date: 26-Feb-2023
Publisher: Radiation Research Society
Citation: Valerie Swee Ting Goh, Yohei Fujishima, Ryo Nakayama, Kai Takebayashi, Mitsuaki A. Yoshida, Kosuke Kasai, Kentaro Ariyoshi, Tomisato Miura (2023-02-26). Manual Scoring with Shortened 48 h Cytokinesis-Block Micronucleus Assay Feasible for Triage in the Event of a Mass-Casualty Radiation Accident. Radiation Research 199 (4) : 385-395. ScholarBank@NUS Repository.
Abstract: The cytokinesis-block micronucleus (CBMN) assay in cytogenetic biodosimetry uses micronucleus (MN) frequency scored in binucleated cells (BNCs) to estimate ionizing radiation dose exposed. Despite the faster and simpler MN scoring, CBMN assay is not commonly recommended in radiation mass-casualty triage as human peripheral blood is typically cultured for 72 h. Furthermore, CBMN assay evaluation in triage often uses high-throughput scoring with expensive and specialized equipment. In this study, we evaluated the feasibility of a low-cost method of manual MN scoring on Giemsa-stained slides in shortened 48 h cultures for triage. Both whole blood and human peripheral blood mononuclear cell cultures were compared for different culture periods and Cyt-B treatment [48 h (24 h at Cyt-B); 72 h (24 h at Cyt-B); 72 h (44 h at Cyt-B)]. Three donors (26-year-old female, 25-year-old male, 29-year-old male) were used for dose-response curve construction with radiation-induced MN/BNC. Another 3 donors (23-year-old female, 34-year-old male, 51-year-old male) were used for triage and conventional dose estimation comparison after 0, 2 and 4 Gy X-ray exposure. Our results showed that despite lower percentage of BNC in 48 h than 72 h cultures, sufficient BNCs were obtained for MN scoring. Triage dose estimates of 48 h cultures were obtained in 8 min in non-exposed donors, and 20 min in 2 or 4 Gy exposed donors with manual MN scoring. One hundred BNCs could be scored for high doses instead of 200 BNCs for triage. Furthermore, observed triage MN distribution could be preliminarily used to differentiate 2 and 4 Gy samples. The number of BNCs scored (triage or conventional) also did not affect dose estimation. Dose estimates in 48 h cultures were also mostly within ±0.5 Gy of actual doses, thus showing the feasibility of manual MN scoring in the shortened CBMN assay for radiological triage applications.
Source Title: Radiation Research
ISSN: 1938-5404
DOI: 10.1667/RADE-22-00191.1
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