The immunohistochemical streptavidin-peroxidase (SP) method was used to select breast cancer tissue that was punctured or resected during surgery. According to the guidelines, two pathologists independently read the radiographs in a double-blind manner to observe the ER, PR, and HER2 expression. The positive ER and PR staining was localized in the nucleus, showing brown particles. The positive HER2 staining was localized in the cytoplasm or cell membrane and appeared as brown granules. The ER and PR receptor expression criteria were positive, negative, or undecipherable. (1) Positive: The proportion of tumor cells with positive staining in the whole section to all tumor cells was evaluated. When ≥1% of tumor nuclei showed different degrees of staining, this was considered positive. (2) Negative: Approximately <1% of the tumor nuclei in the whole section showed varying degrees of staining or no staining at all. Negative results had to be determined on the basis of good staining of internal and external controls. In the case of ER–/PR+, repeated ER and PR tests were needed to exclude ER false-negative or PR false-positive cases.

In the HER2 test procedure, breast cancer specimens could usually be tested by Immunohistochemistry (IHC) first. HER2 positive for IHC 3+ and HER2 negative for IHC 0 and 1+. IHC 2 + the further use of the method of in situ hybridization (FISH) for HER2 gene amplification state detection and can also select different organizations piece to test or send other lab for testing. If the IHC appeared to be inconsistent with the results of the in situ hybridization detection of the cases, multidisciplinary discussion was suggested, the reason was analyzed, and corresponding treatment strategies were formulated.

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