Principles of Analytic Validation of Immunohistochemical Assays: Guideline From the CAP Pathology and Laboratory Quality Center

Patrick L. Fitzgibbons, Linda A. Bradley, Lisa A. Fatheree, Randa Alsabeh, Regan S. Fulton, Jeffrey D. Goldsmith, Thomas S. Haas, Rouzan G. Karabakhtsian, Patti A. Loykasek, Monna J. Marolt, Steven S. Shen, Anthony T. Smith, and Paul E. Swanson (2014) Principles of Analytic Validation of Immunohistochemical Assays: Guideline From the College of American Pathologists Pathology and Laboratory Quality Center. Archives of Pathology & Laboratory Medicine In-Press.

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Principles of Analytic Validation of Immunohistochemical Assays: Guideline From the College of American Pathologists Pathology and Laboratory Quality Center

Patrick L. Fitzgibbons , MD; Linda A. Bradley , PhD; Lisa A. Fatheree , BS, SCT(ASCP); Randa Alsabeh , MD; Regan S. Fulton , MD, PhD; Jeffrey D. Goldsmith , MD; Thomas S. Haas , DO; Rouzan G. Karabakhtsian , MD, PhD; Patti A. Loykasek , HT(ASCP); Monna J. Marolt , MD; Steven S. Shen , MD, PhD; Anthony T. Smith , MLS; Paul E. Swanson , MD

From the Department of Pathology, St. Jude Medical Center, Fullerton, California (Dr Fitzgibbons); the Department of Pathology and Laboratory Medicine, Women & Infants Hospital/Brown University, Providence, Rhode Island (Dr Bradley); the College of American Pathologists, Northfield, Illinois (Ms Fatheree and Mr Smith); the Department of Pathology, Kaiser Permanente - Los Angeles Medical Center, Los Angeles, California (Dr Alsabeh); PhenoPath Laboratories, Seattle, Washington (Dr Fulton); the Department of Pathology, Beth Israel Deaconess Medical Center, Boston, Massachusetts (Dr Goldsmith); the Department of Pathology, Mercy Hospital, Janesville, Wisconsin (Dr Haas); the Department of Pathology, Montefiore Medical Center, New York, New York (Dr Karabakhtsian); Regional Medical Laboratory, St John's Medical Center, Tulsa, Oklahoma (Ms Loykasek); the Department of Pathology, University of Minnesota Medical Center, Fairview, Minneapolis (Dr Marolt); the Department of Pathology, The Methodist Hospital, Houston, Texas (Dr Shen); and the Department of Pathology, University of Washington Medical Center, Seattle (Dr Swanson).

Context.— Laboratories must validate all assays before they can be used to test patient specimens, but currently there are no evidence-based guidelines regarding validation of immunohistochemical assays.

Objective.— To develop recommendations for initial analytic validation and revalidation of immunohistochemical assays.

Design.— The College of American Pathologists Pathology and Laboratory Quality Center convened a panel of pathologists and histotechnologists with expertise in immunohistochemistry to develop validation recommendations. A systematic evidence review was conducted to address key questions. Electronic searches identified 1463 publications, of which 126 met inclusion criteria and were extracted. Individual publications were graded for quality, and the key question findings for strength of evidence. Recommendations were derived from strength of evidence, open comment feedback, and expert panel consensus.

Results.— Fourteen guideline statements were established to help pathology laboratories comply with validation and revalidation requirements for immunohistochemical assays.

Conclusions.— Laboratories must document successful analytic validation of all immunohistochemical tests before applying to patient specimens. The parameters for cases included in validation sets, including number, expression levels, fixative and processing methods, should take into account intended use and should be sufficient to ensure that the test accurately measures the analyte of interest in specimens tested in that laboratory. Recommendations are also provided for confirming assay performance when there are changes in test methods, reagents, or equipment.