Date of Completion

Winter 12-8-2017

Thesis Advisor(s)

Judy Brown, PhD. CG, MB(ASCP); Melissa L. Gorman, CG(ASCP)

Honors Major

Diagnostic Genetic Sciences

Disciplines

Genetics | Genomics

Abstract

Abbott Molecular recently developed the rapid Vysis IntelliFISH Hybridization Buffer for use in fluorescence in situ hybridization (FISH) assays. The hybridization step in a standard FISH assay requires an overnight incubation, thus the turnaround time for a patient result is at least 24 hours. The IntelliFISH buffer was designed to reduce hybridization time of probes to target DNA to about two hours, allowing the entire assay to be performed in a single day. A review of methodologies and commercial products has highlighted the importance of hybridization kinetics to a successful FISH assay. The purpose of the research was to investigate whether the IntelliFISH buffer is an appropriate reagent for producing high quality FISH results in a single day within a clinical laboratory environment. A method comparison experimental approach was used to evaluate signal intensity and hybridization specificity on a subjective scale with outlined criteria for each scale score. The IntelliFISH hybridization buffer was tested on a total of fifty cases representing direct processed chorionic villus, formalin-fixed paraffin embedded (FFPE) tissue, peripheral blood and bone marrow samples using a variety of probes, and it produced varied results dependent on sample type and probe type. This research was sufficient for validation of the Vysis IntelliFISH Hybridization Buffer. Since the conclusion of the study, the BWH cytogenetics laboratory has started to edit the SOPs for some routine FISH studies to include a rapid hybridization. Faster turnaround times will provide patients with numerous benefits and will improve patient and physician satisfaction overall.

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