Automated Bright-Field Dual-Color In Situ Hybridization for MDM2: Interobserver Reproducibility and Correlation With Fluorescence In Situ Hybridization in a Series of Soft Tissue Consults.
BACKGROUND
-Atypical lipomatous tumors/well-differentiated liposarcomas contain alterations in the 12q13-15 region resulting in amplification of MDM2 and nearby genes. Identifying MDM2 amplification is a useful ancillary test, as the histologic mimics of atypical lipomatous tumors/well-differentiated liposarcomas have consistently shown a lack of MDM2 amplification.


OBJECTIVE
-To assess the interobserver reproducibility of a bright-field assay for MDM2 amplification (dual-color, dual-hapten in situ hybridization [DDISH]) among reviewers with varying degrees of experience with the assay and to assess the concordance of MDM2 DDISH with MDM2 fluorescence in situ hybridization (FISH).


METHODS
-In total, 102 cases were assessed in parallel for MDM2 by FISH and DDISH. MDM2 amplification was defined as an MDM2 to chromosome 12 ratio of 2.0 or greater, whereas an MDM2 to chromosome 12 ratio of less than 2 was nonamplified. Fluorescence in situ hybridization was scored in the routine clinical laboratory and DDISH was evaluated by 3 different pathologists blinded to the final diagnosis and FISH results.


RESULTS
-Fluorescence in situ hybridization categorized 27 cases (26%) as MDM2 amplified and 75 cases (74%) as nonamplified; the consensus DDISH diagnosis was 98% concordant with FISH. Agreement between MDM2 DDISH by each reviewer and MDM2 FISH was highly concordant (99%, 98%, and 98%, respectively, for reviewers 1, 2 and 3). The κ agreement of the 3 reviewers scoring DDISH was excellent (κ = 0.949, 0.95, and 0.95, respectively, for reviewers 1, 2, and 3).


CONCLUSIONS
-This study highlights excellent concordance between DDISH and FISH in MDM2 copy number assessment. Moreover, excellent interobserver reproducibility of the DDISH assay was found among reviewers with varying levels of experience evaluating bright-field assays.
From the Departments of Anatomic Pathology (Drs Zhang and Goldblum) and Molecular Pathology (Mr Lanigan and Dr Tubbs), Robert J. Tomsich Pathology and Laboratory Medicine Institute, Cleveland Clinic, Cleveland, Ohio; and the Department of Pathology, Huntsman Cancer Center, University of Utah, Salt Lake City (Dr Downs-Kelly).
Gloria
G
Zhang
Christopher P
CP
Lanigan
John R
JR
Goldblum
Raymond R
RR
Tubbs
Erinn
E
Downs-Kelly
D002881:Chromosomes, Human, Pair 12 / Q000235:genetics; D005784:Gene Amplification; D006801:Humans; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D008067:Lipoma / Q000175:diagnosis / Q000235:genetics*; D008080:Liposarcoma / Q000175:diagnosis / Q000235:genetics*; D015588:Observer Variation; D051736:Proto-Oncogene Proteins c-mdm2 / Q000235:genetics*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
C497206:MDM2 protein, human; D051736:Proto-Oncogene Proteins c-mdm2