High-throughput sample processing and sample management; the functional evolution of classical cytogenetic assay towards automation

• Published in: Mutation research. Genetic toxicology and environmental mutagenesis Vol. 793; pp. 132 - 141
• Main Authors: Ramakumar, Adarsh, Subramanian, Uma, Prasanna, Pataje G.S.
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.11.2015; Elsevier BV
• Subjects: Automation; Automation, Laboratory - standards; Bioassays; Biodosimetry; Bioinformatics; Chromosome Aberrations; Chromosomes; Chromosomes, Human - genetics; Chromosomes, Human - radiation effects; Cytogenetic Analysis - instrumentation; Cytogenetic Analysis - methods; Cytogenetic laboratory; Cytogenetics; Dicentric assay; Dosimetry; High-Throughput Screening Assays - instrumentation; High-Throughput Screening Assays - methods; Humans; International regulations; Laboratory information system; Quality assurance; Quality control; Radiation exposure assessment; Radiation mass casualty triage; Radiometry - instrumentation; Radiometry - methods; Sample preparation; LIMS; Automation; Biodosimetry; DCA; SOP; IAEA; Radiation mass casualty triage; Radiation exposure assessment; IR; Cytogenetic laboratory; Laboratory information system; Dicentric assay
• ISSN: 1383-5718; 1879-3592; 1879-3592
• DOI: 10.1016/j.mrgentox.2015.07.011

•Functional evolution of the Dicentric Chromosome Assay (DCA).•Transition of DCA from bench to automated platforms to increase throughput.•Currently existing gaps, limitations, challenges, and solutions for improvements.•DCA application in radiation mass-casualty and path forward for automated cytogenetics. High-throughput individual diagnostic dose assessment is essential for medical management of radiation-exposed subjects after a mass casualty. Cytogenetic assays such as the Dicentric Chromosome Assay (DCA) are recognized as the gold standard by international regulatory authorities. DCA is a multi-step and multi-day bioassay. DCA, as described in the IAEA manual, can be used to assess dose up to 4–6 weeks post-exposure quite accurately but throughput is still a major issue and automation is very essential. The throughput is limited, both in terms of sample preparation as well as analysis of chromosome aberrations. Thus, there is a need to design and develop novel solutions that could utilize extensive laboratory automation for sample preparation, and bioinformatics approaches for chromosome-aberration analysis to overcome throughput issues. We have transitioned the bench-based cytogenetic DCA to a coherent process performing high-throughput automated biodosimetry for individual dose assessment ensuring quality control (QC) and quality assurance (QA) aspects in accordance with international harmonized protocols. A Laboratory Information Management System (LIMS) is designed, implemented and adapted to manage increased sample processing capacity, develop and maintain standard operating procedures (SOP) for robotic instruments, avoid data transcription errors during processing, and automate analysis of chromosome-aberrations using an image analysis platform. Our efforts described in this paper intend to bridge the current technological gaps and enhance the potential application of DCA for a dose-based stratification of subjects following a mass casualty. This paper describes one such potential integrated automated laboratory system and functional evolution of the classical DCA towards increasing critically needed throughput.