Preclinical validation of fluorescence in situ hybridization assays for clinical practice

Anne E. Wiktor, Daniel L. Van Dyke, Peggy J. Stupca, Rhett P. Ketterling, Erik C Thorland, Brandon M. Shearer, Stephanie R. Fink, Kimberly J. Stockero, Jason R. Majorowicz, Gordon W. Dewald

Research output: Contribution to journalArticle

70 Citations (Scopus)

Abstract

Purpose: Validation of fluorescence in situ hybridization assays is required before using them in clinical practice. Yet, there are few published examples that describe the validation process, leading to inconsistent and sometimes inadequate validation practices. The purpose of this article is to describe a broadly applicable preclinical validation process. Methods: Validation is performed using four consecutive experiments. The Familiarization experiment tests probe performance on metaphase cells to measure analytic sensitivity and specificity for normal blood specimens. The Pilot Study tests a variety of normal and abnormal specimens, using the intended tissue type, to set a preliminary normal cutoff and establish the analytic sensitivity. The Clinical Evaluation experiment tests these parameters in a series of normal and abnormal specimens to simulate clinical practice, establish the normal cutoff and abnormal reference ranges, and finalize the standard operating procedure. The Precision experiment measures the reproducibility of the new assay over 10 consecutive working days. To illustrate documentation and analysis of data with this process, the results for a new assay to detect fusion of IGH and BCL3 associated with t(14;19)(q32; q13.3) in lymphoproliferative disorders are provided in this report. Results: These four experiments determine the analytic sensitivity and specificity, normal values, precision, and reportable reference ranges for validation of the new test. Conclusion: This report describes a method for preclinical validation of fluorescence in situ hybridization studies of metaphase cells and interphase nuclei using commercial or home brew probes.

Original languageEnglish (US)
Pages (from-to)16-23
Number of pages8
JournalGenetics in Medicine
Volume8
Issue number1
DOIs
StatePublished - Jan 2006

Fingerprint

Fluorescence In Situ Hybridization
Reference Values
Metaphase
Sensitivity and Specificity
Lymphoproliferative Disorders
Interphase
Cell Nucleus
Documentation

Keywords

  • FISH validation
  • Interphase FISH
  • Metaphase FISH

ASJC Scopus subject areas

  • Genetics(clinical)
  • Genetics

Cite this

Wiktor, A. E., Van Dyke, D. L., Stupca, P. J., Ketterling, R. P., Thorland, E. C., Shearer, B. M., ... Dewald, G. W. (2006). Preclinical validation of fluorescence in situ hybridization assays for clinical practice. Genetics in Medicine, 8(1), 16-23. https://doi.org/10.1097/01.gim.0000195645.00446.61

Preclinical validation of fluorescence in situ hybridization assays for clinical practice. / Wiktor, Anne E.; Van Dyke, Daniel L.; Stupca, Peggy J.; Ketterling, Rhett P.; Thorland, Erik C; Shearer, Brandon M.; Fink, Stephanie R.; Stockero, Kimberly J.; Majorowicz, Jason R.; Dewald, Gordon W.

In: Genetics in Medicine, Vol. 8, No. 1, 01.2006, p. 16-23.

Research output: Contribution to journalArticle

Wiktor, AE, Van Dyke, DL, Stupca, PJ, Ketterling, RP, Thorland, EC, Shearer, BM, Fink, SR, Stockero, KJ, Majorowicz, JR & Dewald, GW 2006, 'Preclinical validation of fluorescence in situ hybridization assays for clinical practice', Genetics in Medicine, vol. 8, no. 1, pp. 16-23. https://doi.org/10.1097/01.gim.0000195645.00446.61
Wiktor, Anne E. ; Van Dyke, Daniel L. ; Stupca, Peggy J. ; Ketterling, Rhett P. ; Thorland, Erik C ; Shearer, Brandon M. ; Fink, Stephanie R. ; Stockero, Kimberly J. ; Majorowicz, Jason R. ; Dewald, Gordon W. / Preclinical validation of fluorescence in situ hybridization assays for clinical practice. In: Genetics in Medicine. 2006 ; Vol. 8, No. 1. pp. 16-23.
@article{db913f192cae46389cdf37e1f575ce90,
title = "Preclinical validation of fluorescence in situ hybridization assays for clinical practice",
abstract = "Purpose: Validation of fluorescence in situ hybridization assays is required before using them in clinical practice. Yet, there are few published examples that describe the validation process, leading to inconsistent and sometimes inadequate validation practices. The purpose of this article is to describe a broadly applicable preclinical validation process. Methods: Validation is performed using four consecutive experiments. The Familiarization experiment tests probe performance on metaphase cells to measure analytic sensitivity and specificity for normal blood specimens. The Pilot Study tests a variety of normal and abnormal specimens, using the intended tissue type, to set a preliminary normal cutoff and establish the analytic sensitivity. The Clinical Evaluation experiment tests these parameters in a series of normal and abnormal specimens to simulate clinical practice, establish the normal cutoff and abnormal reference ranges, and finalize the standard operating procedure. The Precision experiment measures the reproducibility of the new assay over 10 consecutive working days. To illustrate documentation and analysis of data with this process, the results for a new assay to detect fusion of IGH and BCL3 associated with t(14;19)(q32; q13.3) in lymphoproliferative disorders are provided in this report. Results: These four experiments determine the analytic sensitivity and specificity, normal values, precision, and reportable reference ranges for validation of the new test. Conclusion: This report describes a method for preclinical validation of fluorescence in situ hybridization studies of metaphase cells and interphase nuclei using commercial or home brew probes.",
keywords = "FISH validation, Interphase FISH, Metaphase FISH",
author = "Wiktor, {Anne E.} and {Van Dyke}, {Daniel L.} and Stupca, {Peggy J.} and Ketterling, {Rhett P.} and Thorland, {Erik C} and Shearer, {Brandon M.} and Fink, {Stephanie R.} and Stockero, {Kimberly J.} and Majorowicz, {Jason R.} and Dewald, {Gordon W.}",
year = "2006",
month = "1",
doi = "10.1097/01.gim.0000195645.00446.61",
language = "English (US)",
volume = "8",
pages = "16--23",
journal = "Genetics in Medicine",
issn = "1098-3600",
publisher = "Lippincott Williams and Wilkins",
number = "1",

}

TY - JOUR

T1 - Preclinical validation of fluorescence in situ hybridization assays for clinical practice

AU - Wiktor, Anne E.

AU - Van Dyke, Daniel L.

AU - Stupca, Peggy J.

AU - Ketterling, Rhett P.

AU - Thorland, Erik C

AU - Shearer, Brandon M.

AU - Fink, Stephanie R.

AU - Stockero, Kimberly J.

AU - Majorowicz, Jason R.

AU - Dewald, Gordon W.

PY - 2006/1

Y1 - 2006/1

N2 - Purpose: Validation of fluorescence in situ hybridization assays is required before using them in clinical practice. Yet, there are few published examples that describe the validation process, leading to inconsistent and sometimes inadequate validation practices. The purpose of this article is to describe a broadly applicable preclinical validation process. Methods: Validation is performed using four consecutive experiments. The Familiarization experiment tests probe performance on metaphase cells to measure analytic sensitivity and specificity for normal blood specimens. The Pilot Study tests a variety of normal and abnormal specimens, using the intended tissue type, to set a preliminary normal cutoff and establish the analytic sensitivity. The Clinical Evaluation experiment tests these parameters in a series of normal and abnormal specimens to simulate clinical practice, establish the normal cutoff and abnormal reference ranges, and finalize the standard operating procedure. The Precision experiment measures the reproducibility of the new assay over 10 consecutive working days. To illustrate documentation and analysis of data with this process, the results for a new assay to detect fusion of IGH and BCL3 associated with t(14;19)(q32; q13.3) in lymphoproliferative disorders are provided in this report. Results: These four experiments determine the analytic sensitivity and specificity, normal values, precision, and reportable reference ranges for validation of the new test. Conclusion: This report describes a method for preclinical validation of fluorescence in situ hybridization studies of metaphase cells and interphase nuclei using commercial or home brew probes.

AB - Purpose: Validation of fluorescence in situ hybridization assays is required before using them in clinical practice. Yet, there are few published examples that describe the validation process, leading to inconsistent and sometimes inadequate validation practices. The purpose of this article is to describe a broadly applicable preclinical validation process. Methods: Validation is performed using four consecutive experiments. The Familiarization experiment tests probe performance on metaphase cells to measure analytic sensitivity and specificity for normal blood specimens. The Pilot Study tests a variety of normal and abnormal specimens, using the intended tissue type, to set a preliminary normal cutoff and establish the analytic sensitivity. The Clinical Evaluation experiment tests these parameters in a series of normal and abnormal specimens to simulate clinical practice, establish the normal cutoff and abnormal reference ranges, and finalize the standard operating procedure. The Precision experiment measures the reproducibility of the new assay over 10 consecutive working days. To illustrate documentation and analysis of data with this process, the results for a new assay to detect fusion of IGH and BCL3 associated with t(14;19)(q32; q13.3) in lymphoproliferative disorders are provided in this report. Results: These four experiments determine the analytic sensitivity and specificity, normal values, precision, and reportable reference ranges for validation of the new test. Conclusion: This report describes a method for preclinical validation of fluorescence in situ hybridization studies of metaphase cells and interphase nuclei using commercial or home brew probes.

KW - FISH validation

KW - Interphase FISH

KW - Metaphase FISH

UR - http://www.scopus.com/inward/record.url?scp=33646594424&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33646594424&partnerID=8YFLogxK

U2 - 10.1097/01.gim.0000195645.00446.61

DO - 10.1097/01.gim.0000195645.00446.61

M3 - Article

C2 - 16418595

AN - SCOPUS:33646594424

VL - 8

SP - 16

EP - 23

JO - Genetics in Medicine

JF - Genetics in Medicine

SN - 1098-3600

IS - 1

ER -