Dr. Patrick R. Gonzales is a Clinical Assistant Professor, Co-Director of the Cytogenetics Laboratory, and Clinical Consultant for the Clinical Molecular Oncology Laboratory, in the Department of Pathology and Laboratory Medicine at the University of Kansas Medical Center. Building upon his prior experience in clinical cytogenetics at Mayo Clinic, and molecular genetics at Washington University in St. Louis, he completed his training at UAB in clinical cytogenetics, molecular genetics and genomics to properly interpret mechanisms of genetic disease. He is ABMGG certified in Clinical Cytogenetics and Genomics, as well as Clinical Molecular Genetics and Genomics.
Patrick R. Gonzales, Traci Troyer, Claire Pierson, Hallie Craycraft, Shivani Golem
University of Kansas Medical Center, Kansas City, KS, USA
Automation of FISH analysis is an option to increase laboratory throughput. The BioView Duet-3 System was validated for clinical use in FISH analysis of chronic lymphocytic leukemia (CLL) specimens. CLL is the most common hematological neoplasia in Western countries. FISH analysis remains the gold standard for assessment of genomic abnormalities in CLL. The CLL FISH panel included probes for deletions of 6q (MYB), 11q (ATM), 13q14.3 and 17p (TP53), and trisomy 12. Twenty-one specimens (Ten new diagnoses and 11 follow-up) were analyzed in parallel with BioView and standard manual analysis. Single observer scored the output of BioView and compared results with manual scoring of 200 or 500 interphase nuclei. Second scorer results were assessed for interobserver variability. Normal cutoffs for 6q, 11q, and +12 remained similar to manual cutoffs, while the cutoffs were higher (+2.8~4.4%) for 13q and 17p probes for BioView. Concordance was 0.99 between BioView and manual scoring (1 FP/105 total assays). Interobserver concordance was 100%. The concordance was 100% for probes 6q, 11q, +12 and 17p. In one FP 13q case, along with the concordant 1R2G signal pattern, an additional 1R1G (-13) signal pattern was detected which was slightly above the cutoff value (<5.0% vs 5.5%). Overall, the BioView system showed high concordance. Other advantages included reduced hands-on use of microscopy, with raw images providing for lab directors to judge scored signal patterns, simplifying test review and analysis. Limitations include manual microscope review of the abnormal signal patterns which are slightly above the cutoff values before.