PMID- 37168512
OWN - NLM
STAT- PubMed-not-MEDLINE
LR  - 20230514
IS  - 1936-2625 (Electronic)
IS  - 1936-2625 (Linking)
VI  - 16
IP  - 4
DP  - 2023
TI  - Validation of MYC and BCL6 rapid break apart digital fluorescence in situ 
      hybridization assays for clinical use.
PG  - 76-85
AB  - OBJECTIVE: Detection of gene rearrangements in MYC (a family of regulator genes 
      and proto-oncogenes) and human B-cell lymphoma 6 (BCL6) using fluorescence in 
      situ hybridization (FISH) are important in the evaluation of lymphomas, in 
      particular diffuse large B-cell lymphoma (DLBCL) and Burkitt lymphoma. Our 
      current clinical MYC and BCL6 FISH workflow involves an overnight hybridization 
      of probes with digital analysis using the GenASIs Scan and Analysis instrument 
      (Applied Spectral Imaging). In order to improve assay turnaround time SureFISH 
      probes were validated to reduce the hybridization time from 16 hours down to 1.5 
      hours. METHODS: Validation was a four-phase process involving initial development 
      of the assays by testing new probes in a manual protocol, and cytogenetic studies 
      to confirm the probe specificity, sensitivity, and localization. In the next 
      phase, the assays were validated as a manual assay. The third phase involved 
      development of the digital FISH assays by testing and optimizing the GenASIs Scan 
      and Analysis instrument. In the final phase, the digital FISH assays were 
      validated. RESULTS: Cytogenetic studies confirmed 100% probe 
      sensitivity/specificity, and localization patterns. Negative reference range 
      cutoffs calculated from 20 normal lymph nodes using the inverse of the beta 
      cumulative probability density function (Excel BETAINV calculation) were 11% 
      inclusive for both manual and digital MYC and BCL6 assays. There was 100% 
      concordance between the manual and digital methods. The shortened hybridization 
      time decreased the overall workflow time by 14.5 hours. CONCLUSIONS: This study 
      validates the use of the SureFISH MYC and BCL6 probes on formalin fixed paraffin 
      embedded (FFPE) tissue sections using a hybridization time of 1.5 hours that 
      shortened the overall workflow by 14.5 hours. The process described also provides 
      a standardized framework for validating digital FISH assays in the future.
CI  - IJCEP Copyright (c) 2023.
FAU - Liew, Michael
AU  - Liew M
AD  - ARUP Institute for Clinical and Experimental Pathology(R) Salt Lake, UT 84108, USA.
FAU - Rowe, Leslie
AU  - Rowe L
AD  - ARUP Institute for Clinical and Experimental Pathology(R) Salt Lake, UT 84108, USA.
FAU - Moore, Kristina
AU  - Moore K
AD  - ARUP Institute for Clinical and Experimental Pathology(R) Salt Lake, UT 84108, USA.
FAU - Aston, Emily
AU  - Aston E
AD  - ARUP Institute for Clinical and Experimental Pathology(R) Salt Lake, UT 84108, USA.
FAU - O'Brien, Kathryn
AU  - O'Brien K
AD  - ARUP Institute for Clinical and Experimental Pathology(R) Salt Lake, UT 84108, USA.
FAU - Longhurst, Maria
AU  - Longhurst M
AD  - ARUP Institute for Clinical and Experimental Pathology(R) Salt Lake, UT 84108, USA.
FAU - Kenney, Jason
AU  - Kenney J
AD  - ARUP Institute for Clinical and Experimental Pathology(R) Salt Lake, UT 84108, USA.
FAU - Priest, Marshall
AU  - Priest M
AD  - ARUP Institute for Clinical and Experimental Pathology(R) Salt Lake, UT 84108, USA.
FAU - Zhou, Wenhua
AU  - Zhou W
AD  - ARUP Institute for Clinical and Experimental Pathology(R) Salt Lake, UT 84108, USA.
FAU - Wilcock, Diane
AU  - Wilcock D
AD  - ARUP Institute for Clinical and Experimental Pathology(R) Salt Lake, UT 84108, USA.
FAU - Rets, Anton
AU  - Rets A
AD  - ARUP Institute for Clinical and Experimental Pathology(R) Salt Lake, UT 84108, USA.
AD  - Department of Pathology, University of Utah School of Medicine Salt Lake City, UT 
      84132, USA.
FAU - Miles, Rodney
AU  - Miles R
AD  - ARUP Institute for Clinical and Experimental Pathology(R) Salt Lake, UT 84108, USA.
AD  - Department of Pathology, University of Utah School of Medicine Salt Lake City, UT 
      84132, USA.
LA  - eng
PT  - Journal Article
DEP - 20230415
PL  - United States
TA  - Int J Clin Exp Pathol
JT  - International journal of clinical and experimental pathology
JID - 101480565
PMC - PMC10165167
OTO - NOTNLM
OT  - GenASIs analysis system
OT  - digital pathology
COIS- None.
EDAT- 2023/05/12 01:07
MHDA- 2023/05/12 01:08
PMCR- 2023/04/15
CRDT- 2023/05/11 19:22
PHST- 2022/11/29 00:00 [received]
PHST- 2023/03/29 00:00 [accepted]
PHST- 2023/05/12 01:08 [medline]
PHST- 2023/05/12 01:07 [pubmed]
PHST- 2023/05/11 19:22 [entrez]
PHST- 2023/04/15 00:00 [pmc-release]
PST - epublish
SO  - Int J Clin Exp Pathol. 2023 Apr 15;16(4):76-85. eCollection 2023.