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Related Experiment Video

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An Automated Method to Perform The In Vitro Micronucleus Assay using Multispectral Imaging Flow Cytometry
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RENEB Inter-Laboratory Comparison 2021: The Cytokinesis-Block Micronucleus Assay.

A Vral1, D Endesfelder2, J Balázs3

  • 1Faculty of Medicine and Health Sciences, Radiobiology Research Unit, Universiteit Gent, Gent, Belgium.

Radiation Research
|April 14, 2023
PubMed
Summary
This summary is machine-generated.

The cytokinesis-block micronucleus assay effectively classified radiation doses in 88% of cases during a large-scale emergency simulation. Semi-automated scoring demonstrated the highest accuracy and reliability for rapid biodosimetry in radiation accidents.

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Area of Science:

  • Radiation Biology
  • Cytogenetics
  • Biodosimetry

Background:

  • Large-scale radiation accidents necessitate rapid biodosimetry for effective triage and medical management.
  • Inter-laboratory comparisons are crucial for validating biodosimetric assays under simulated emergency conditions.
  • The cytokinesis-block micronucleus assay is a key cytogenetic method for assessing radiation exposure.

Purpose of the Study:

  • To evaluate the performance of the cytokinesis-block micronucleus assay in a simulated large-scale radiation accident scenario.
  • To assess the accuracy and reliability of dose estimations using different scoring modes (manual, semi-automated, automated) in triage biodosimetry.
  • To determine the effectiveness of the micronucleus assay in classifying radiation doses into clinically relevant triage categories.

Main Methods:

  • Fourteen laboratories participated in an inter-laboratory comparison exercise using the cytokinesis-block micronucleus assay.
  • Test samples were exposed to blinded doses of X-rays (0, 1.2, and 3.5 Gy) and analyzed in triage mode.
  • Dose estimations were performed by scoring binucleated cells (500 manually, 1000 automated/semi-automated) and comparing results to calibration curves.

Main Results:

  • The micronucleus assay correctly classified samples into clinical triage categories (low, medium, high dose) in 88% of cases.
  • Semi-automated scoring achieved 100% accuracy in classification and excellent inter-laboratory agreement (Fleiss' kappa = 100%).
  • Manual scoring showed 73% correct classification into dose intervals, while automated scoring was less reliable (67% correct classification, Fleiss' kappa = 53%). A systematic shift towards higher dose estimations was observed for irradiated samples.

Conclusions:

  • The cytokinesis-block micronucleus assay is a reliable method for rapid biodosimetry in radiation emergencies.
  • Semi-automated scoring offers the highest accuracy and reliability, followed by manual scoring, for triage biodosimetry.
  • Reducing manually scored cells to 200 binucleated cells can maintain the accuracy of the rapid triage modality.