BACKGROUNDSome evidence suggests a role for HER‐2/neu overexpression in prostate carcinoma progression. Reported rates of HER‐2/neu overexpression in patients with prostate carcinoma vary greatly.METHODSThe authors studied radical prostatectomy specimens from 38 patients who had biochemical failure after undergoing radical prostatectomy for prostate carcinoma. Immunohistochemistry for HER‐2/neu overexpression using the HercepTest kit (Dako Corporation, Carpenteria, CA) was employed. Two different antigen‐retrieval techniques were used: 1) the standard U.S. Food and Drug Administration (FDA)‐approved HercepTest assay and 2) a modified HercepTest, which employed an alkaline citrate buffer, pH 9.0, for antigen retrieval and a 1‐hour primary antibody incubation time. The level of HER‐2/neu expression was evaluated on a scale from 0 (no staining) to 3+ according to the published guidelines. Fluorescent in situ hybridization for gene amplification was performed on all specimens.RESULTSWith the standard technique, only one specimen had 2+ staining, and no specimens had 3+ staining. With the modified technique, 10 specimens (26%) had 2+ staining, and 9 specimens (24%) had 3+ staining. There was a significant association between the level of HER‐2/neu expression shown with the modified technique and tumor stage (P = 0.03) as well as Gleason grade (P = 0.01). None of the specimens had HER‐2/neu gene amplification.CONCLUSIONSThe authors report a simple modification of the HercepTest that resulted in an increased rate of HER‐2/neu expression, which was correlated with poor‐risk pathologic findings. The findings suggest that adenocarcinoma of the prostate should be evaluated for HER‐2/neu expression with a prostate specific immunohistochemical procedure that differs from the FDA‐approved standard procedure. Cancer 2002;95:1650–5. © 2002 American Cancer Society.DOI 10.1002/cncr.10839