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

Abstract

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.

Tissue heterogeneity contributes to suboptimal precision of WHO 2010 scoring criteria for Ki67 labeling index in a subset of neuroendocrine neoplasms of the pancreascriteria for Ki67 labeling index in a subset of neuroendocrine neoplasms of the pancreas

“…Many parameters are used for analytical validation of biomarker assays, including accuracy, trueness, precision, reproducibility, repeatability, linearity, and robustness [52,54]. Analytical validation of IHC tests has gained attention recently [55]. To analytically validate IHC test, it is necessary to examine its accuracy, analytical sensitivity, analytical specificity, and concordance with a gold standard, as well as its robustness and precision [55].…”

“…Analytical validation of IHC tests has gained attention recently [55]. To analytically validate IHC test, it is necessary to examine its accuracy, analytical sensitivity, analytical specificity, and concordance with a gold standard, as well as its robustness and precision [55]. As there is no reference gold standard test for almost all IHC assays [55,56], therefore documentation of accuracy of IHC test may be problematic.…”

“…To analytically validate IHC test, it is necessary to examine its accuracy, analytical sensitivity, analytical specificity, and concordance with a gold standard, as well as its robustness and precision [55]. As there is no reference gold standard test for almost all IHC assays [55,56], therefore documentation of accuracy of IHC test may be problematic. Validation of IHC assay needs to deal with documentation of pre-analytical, analytical, and post-analytical phases of the test [39,55,57,58].…”

Strict rules are needed for validation of G-protein-coupled receptor immunohistochemical studies in human tissues

“…But it is evident that the adherence to such a strategy by all journals reporting IHC data would be beneficial. Valuable in this regard is also a set of recommendations for the validation of IHC assays in diagnostic settings that have recently been published by the College of American Pathologists [6].…”

“…5 Ideally, one should compare the obtained GPCR-IHC data with the localization of the same GPCR using another antibody recognizing a different epitope of the receptor when such a well-validated antibody is available. 6 It is true for every IHC study that an optimal formalinfixation of the human tissue samples, according to the standard rules of surgical pathology, is mandatory [6].…”