Brightfield triplex immunohistochemistry assay for evaluating the colocalization of the er, pr, and ki-67 biomarkers in cells
AssigneesVentana Medical Systems, Inc., Japanese Foundation for Cancer Research
InventorsRie Horii, Hiro Nitta, Takayuki Ueno
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Abstract
The present disclosure is directed to a triplex immunohistochemical assay for detecting the colocalization of the ER, PR, and Ki-67 biomarkers in cells or cell nuclei. It is believed that the brightfield triplex immunohistochemical assay of the present disclosure may serve as a prognostic assay for ER-positive breast cancer, facilitating the identification of disease-free survivors among ER-positive breast cancer patients treated with hormone therapy.
Claims20
An affinity histochemical or affinity cytochemical method for assessing expression of the PR, ER, and Ki-67 biomarkers in a sample, the method comprising:
- (a) contacting the sample with a human PR biomarker-specific reagent under conditions that permit specific binding of the PR biomarker-specific reagent to the sample;
- (b) contacting the sample with a first set of detection reagents that interact with the human PR biomarker-specific reagent to facilitate deposition of a first brightfield detectable moiety on the sample;
- (c) contacting the sample with a human ER biomarker-specific reagent under conditions that permit specific binding of the ER biomarker-specific reagent to the sample;
- (d) contacting the sample with a second set of detection reagents that interact with the human ER biomarker-specific reagent to facilitate deposition of a second brightfield detectable moiety on the sample;
- (e) contacting the sample with a human Ki-67 biomarker-specific reagent under conditions that permit specific binding of the Ki-67 biomarker-specific reagent to the sample;
- (f) contacting the sample with a third set of detection reagents that interact with the human Ki-67 biomarker-specific reagent to facilitate deposition of a third brightfield detectable moiety on the sample;
- and
- (g) identifying cell nuclei within the sample expressing each of the PR, ER, and Ki-67 biomarkers based on a colocalization of signals from each of the deposited first, second, and third brightfield detectable moieties.
A method of selecting a patient with a breast tumor to receive a hormone therapy, the method comprising:
- (a) affinity histochemically staining a sample derived from the breast tumor with a human PR biomarker-specific reagent under conditions that permit specific binding of the PR biomarker-specific reagent to the sample;
- (b) affinity histochemically staining the sample derived from the breast tumor with a human ER biomarker-specific reagent under conditions that permit specific binding of the ER biomarker-specific reagent to the sample;
- (c) affinity histochemically staining the sample derived from the breast tumor with a human Ki-67 biomarker-specific reagent under conditions that permit specific binding of the Ki-67 biomarker-specific reagent to the sample;
- and
- (d) determining a number of proliferating tumor nuclei that are both ER+ and PR+ within the histochemically stained sample;
- wherein the patient is selected to receive the hormone therapy if a ratio of the determined number of proliferating tumor nuclei that are both ER+ and PR+ to a total number of ER+proliferating tumor nuclei is greater or equal to a predetermined cutoff value.
Dependencies are inferred from the claim text.