JP6599710B2 - Cocktail antibody - Google Patents

Description

  The present invention relates to a cocktail antibody that prevents variation and oversight by a judge in pathological diagnosis.

  Pathological diagnosis is performed not only by morphological observation of biological specimens but also by immunohistochemical staining (Patent Document 1). Immunohistochemical staining is a method in which a specific antigen on a tissue is detected by an antibody that specifically recognizes the antigen, and an antibody that recognizes the specific antigen is reacted with the tissue, and the presence or absence of the reacted antibody is detected. Determine the presence of the antigen. The immunohistochemical staining method includes a direct method in which a dye or enzyme is directly bound to a primary antibody that is a specific antibody against an antigen and an indirect method in which a visualization is performed using a secondary antibody that is an antibody against the primary antibody. It is.

  In pathological diagnosis of breast tumor biopsy material, it is very important to (i) determine whether the tumor is infiltrating or stay in the duct and (ii) determine whether the tumor is benign or malignant. (Patent Document 2). In HE staining, (i) it is difficult to recognize myoepithelial cells, which is evidence that the tumor remains in the breast duct, and (ii) determination of whether the tumor is benign or malignant requires skill.

  In contrast to (i), in order to determine whether the tumor is infiltrating or staying in the breast duct, in addition to the HE-stained slide, cytokeratin 5/6 (CK5 / 6) containing calponin, p63, actin, etc. The presence or absence of myoepithelial cells is confirmed by staining multiple sheets with myoepithelial cell markers. Whether the breast tumor is benign or malignant is determined by HE staining, nuclear atypia, nuclear enlargement, karyotypic irregularity, fission, chromatin increase, degree of deviation from normal construction, etc. In addition to morphological judgment, CK5 / 6, cytokeratin 14 (CK14), high molecular weight keratin staining, malignant tumor cells are not stained, normal epithelial hyperplastic cells are stained, estrogen Receptor (ER) staining or progesterone receptor (PgR) staining is performed, and a large number of malignant tumor cells are stained, and normal epithelial hyperplasia is stained only sporadically. However, CK5 / 6 immunostaining often experiences loss of myoepithelial staining. For this reason, in order to confirm invasive / non-invasive cancer, myoepithelial markers other than CK5 / 6 are often added and stained.

  On the other hand, CK5 / 6, CK14, and high molecular weight keratin staining are frequently used because they can distinguish malignant tumor cells but not normal epithelial hyperplastic cells.

  In addition to the HE staining, CK5 / 6 or CK14, high molecular weight keratin staining, and at least two slides of myoepithelial cell marker staining other than CK5 / 6 such as calponin or p63 or actin are included in the above determination. is necessary.

JP 2003-130866 A No. 2012/141285

  The present invention has been made in view of such problems, and provides a cocktail antibody that is advantageous in terms of time and cost, and that can prevent variation and oversight by a judge in pathological diagnosis. Objective.

  The cocktail antibody according to the present invention is characterized by containing an anti-estrogen receptor antibody (anti-ER antibody) and an anti-progesterone antibody (anti-PgR antibody), which stain physically the same pathological specimen material sections.

  The cocktail antibody according to the present invention stains the same pathological specimen material physically, anti-estrogen receptor antibody (anti-ER antibody), anti-progesterone antibody (anti-PgR antibody) and anti-calponin antibody (anti-calponin antibody) ).

  In addition, the cocktail antibody according to the present invention is an anti-cytokeratin 5/6 antibody (anti-CK5 / 6 antibody) or anti-cytokeratin 5 antibody (anti-CK5 antibody) or anti-stain that stains the same pathological specimen material section. It contains cytokeratin 14 antibody (anti-CK14 antibody) or anti-cytokeratin 5/14 antibody (anti-CK5 / 14 antibody) or anti-high molecular weight cytokeratin antibody (34βE12 antibody) and anti-calponin antibody.

  Conventionally, in order to determine the suitability of hormonal therapy for breast tissue histopathology specimens, immunostaining is performed with one anti-ER antibody and one with anti-PgR antibody, and either ER or PgR is applied for hormonal therapy. However, in the present invention, immunostaining is performed with a cocktail antibody using an anti-ER antibody and an anti-PgR antibody, so that the determination of suitability of hormone therapy can be completed with one staining method. Is advantageous. In addition, the determination of hormone receptors has been performed using two separate sections for ER and PgR. Each has a positive-negative cutoff value, and both ER and PgR are both negative if they are below the cutoff value. However, according to the present invention, since one section is stained with two types of antibodies at the same time, ER and PgR are more important than determination by separate staining. The number of positive cells that must not be counted is counted more, the indication for hormonal therapy is more accurate, and the number of indications increases.

  In the present invention, since immunostaining is performed with a cocktail antibody of anti-ER antibody, anti-PgR antibody and anti-calponin antibody, determination of the suitability of hormone therapy, determination of whether the tumor is infiltrating or staying in the breast duct, and tumor is benign The determination of whether it is malignant or not can be completed simultaneously with one sheet. Since one sheet can be completed simultaneously, it is advantageous in terms of time and economy.

  In the present invention, since anti-CK5 / 6 antibody, anti-CK5 antibody, anti-CK14 antibody, anti-CK5 / 14 antibody, anti-high molecular weight cytokeratin antibody (34βE12 antibody) and cocktail antibody with calponin antibody are used, immunostaining is performed. And the determination of whether the tumor is infiltrating, staying in the breast duct, or whether the tumor is benign or malignant can be completed simultaneously with one sheet. Since one sheet can be completed at the same time, it is advantageous in time and economy.

This is HE staining in cases of non-invasive ductal carcinoma in situ (DCIS) and non-invasive lobular carcinoma (LIS). Hematoxylin staining + (CK5 / 6 1x) in breast DCIS and LCIS cases. Hematoxylin staining + (calponin 1x) in breast DCIS and LCIS cases. Hematoxylin staining + (CK5 / 6 1x) + (calponin 1x) in breast DCIS and LCIS cases. Hematoxylin staining + (CK5 / 6 1 / 2x) + (calponin 1 / 2x) in breast DCIS and LCIS cases. Hematoxylin staining + (CK5 / 6 1 / 4x) + (calponin 1 / 4x) in breast DCIS and LCIS cases. Hematoxylin staining + (CK5 / 6 1 / 8x) + (calponin 1 / 8x) in breast DCIS and LCIS cases. Hematoxylin staining + (ER 1x) in breast DCIS and LCIS cases. Hematoxylin staining + (PgR 1x) in breast DCIS and LCIS cases. Hematoxylin staining + (ER 1x) + (PgR 1x) in breast DCIS and LCIS cases. Hematoxylin staining + (ER 1 / 2x) + (PgR 1 / 2x) in breast DCIS and LCIS cases. Hematoxylin staining + (ER 1 / 4x) + (PgR 1 / 4x) in breast DCIS and LCIS cases. Hematoxylin staining + (ER 1 / 8x) + (PgR 1 / 8x) in breast DCIS and LCIS cases. Hematoxylin staining + (ER 1 / 16x) + (PgR 1 / 16x) in breast DCIS and LCIS cases. Hematoxylin staining + (ER 1x) + (PgR 1x) + (calponin 1x) in breast DCIS and LCIS cases. Hematoxylin staining + (ER 1x) + (PgR 1x) + (calponin 1 / 2x) in breast DCIS and LCIS cases. Hematoxylin staining + (ER 1x) + (PgR 1x) + (calponin 1 / 4x) in breast DCIS and LCIS cases. Hematoxylin staining + (CK14 1 / 2x) + (calponin 1 / 2x) in breast DCIS and LCIS cases. Hematoxylin staining + (CK14 1 / 4x) + (calponin 1 / 4x) in breast DCIS and LCIS cases.

  Hereinafter, embodiments of the present invention will be specifically described with reference to the accompanying drawings. However, the embodiments are for facilitating understanding of the principle of the present invention, and the scope of the present invention is as follows. The present invention is not limited to the embodiments, and other embodiments in which those skilled in the art appropriately replace the configurations of the following embodiments are also included in the scope of the present invention.

  The cocktail antibody according to this embodiment contains an anti-estrogen receptor antibody and an anti-progesterone antibody (ER + PgR) that stains physically identical pathological specimen material sections. Here, the cocktail antibody refers to a product in which two or more kinds of antibodies are mixed.

  Factors that determine the type of breast cancer include: (i) whether cancer cells proliferate in response to female hormone receptors (estrogen receptor (ER) or progesterone receptor (PR)); (ii) cancer cells (Iii) whether cancer cells have a high or low degree of proliferative activity. In (i), when both or both of the estrogen receptor (ER) and progesterone receptor (PR) are positive, the effect of hormone therapy can be expected. According to the present invention, ER and PgR do not overlap. The number of positive cells is counted more, the suitability of this hormone therapy is more accurately determined, and the number of indication cases increases.

  For example, if ER and PgR have a positive / negative cut-off value of less than 10%, if ER: 8% negative and PgR: 8% negative are determined, both are traditionally negative. And hormonal therapy is no longer applied. However, according to the present invention, since one section is stained with two types of antibodies at the same time, there are positive cells in which ER and PgR do not overlap. For example, if determined to be 15%, the hormone receptor is positive. Judgment and indication of hormone therapy. Furthermore, since the cocktail antibody of this combination is completed simultaneously with one sheet, it is advantageous in terms of time and economy.

  Next, the cocktail antibody according to the present embodiment contains an anti-estrogen receptor antibody, an anti-progesterone antibody, and an anti-calponin antibody that stains physically identical pathological specimen material sections (ER + PgR + calponin).

  Calponin staining has a high myeloepithelial labeling rate, making it easy to determine whether the cancer is infiltrating or staying in the breast duct. ER and PgR staining can stain malignant tumor cells but normal epithelial hyperplastic cells. Is stained only sporadic, making it easier to distinguish whether a tumor is benign or malignant. Furthermore, since the cocktail antibody of this combination is completed simultaneously with one sheet, it is advantageous in terms of time and economy.

  Next, the cocktail antibody according to the present embodiment is an anti-cytokeratin 5/6 antibody (anti-CK5 / 6 antibody) or anti-cytokeratin 5 antibody (anti-CK5 antibody) or a staining of physically identical pathological specimen material sections. An anti-cytokeratin 14 antibody (anti-CK14 antibody), an anti-cytokeratin 5/14 antibody (anti-CK5 / 14 antibody) or an anti-high molecular weight cytokeratin antibody (34βE12 antibody) and an anti-calponin antibody are contained.

  Cytokeratin 5 is a basic cytokeratin with a molecular weight of 58 kD expressed in stratified epithelium, transitional epithelium, mixed gland, and mesothelial cells, and cytokeratin 6 is a basic cytokeratin with a molecular weight of 56 kD expressed in squamous cells in the proliferating phase . In CK5 / 6 staining, malignant tumor cells are not stained, normal epithelial hyperplastic cells are stained, and it is possible to distinguish whether the tumor is benign or malignant. Anti-CK5 antibody, anti-CK14 antibody, anti-CK5 / 14 antibody and 34βE12 antibody (recognizing cytokeratin numbers 1, 5, 10, and 14) are also known to show the same staining attitude as anti-CK5 / 6 antibody and are malignant. Tumor cells are not stained and normal epithelial hyperplastic cells are stained, and it is possible to distinguish whether the tumor is benign or malignant. In addition, as described above, calponin staining has a high myeloepithelial labeling rate and makes it easy to determine whether cancer is infiltrating or staying in the breast duct. Furthermore, since the cocktail antibody of this combination is completed simultaneously with one sheet, it is advantageous in terms of time and economy.

  The concentration of the anti-cytokeratin 5/6 antibody is not particularly limited, but is, for example, 0.9 μg / L to 45 mg / L. The concentration of the anti-calponin antibody is not particularly limited, for example, 0.86 μg / L to The concentration of anti-cytokeratin 14 antibody is not particularly limited, but is, for example, 2.1 μg / L to 42 mg / L.

  In mammary tumors, ER, PgR, HER2, CK5 / 6, MIB1 are stained and the hormone receptor is expressed, HER2 protein is overexpressed (HER2 gene is amplified), luminal A, luminal B , HER2 overexpressing type, triple negative (basal-like carcinoma) profiling is determined. According to the cocktail antibody according to the present embodiment described above, more accurate determination can be made with three (ER + PgR + calponin), HER2, and MIB1. If it is triple negative, (CK5 / 6 + calponin) may be added to confirm whether it is basal-like carcinoma. Or (ER + PgR), HER2, (CK5 / 6 + calponin), MIB1 can be used. In the case of triple negative, the determination of basal-like carcinoma is completed without additional immunostaining. In the biopsy of breast tumors, in addition to the HE specimen, routine immunostaining with (CK5 / 6 + calponin) or (ER + PgR + calponin) cocktail antibody can be used to determine whether the tumor is invasive or milk A single additional staining can be used to determine whether it remains in the duct or whether the tumor is benign or malignant.

Dyeing was performed under various conditions. The procedure was as shown below.
1. Deparaffinization
(i) Dry sliced section slides.
(ii) Melting paraffin in a 70 ° C incubator (over 10 minutes)
(iii) Using Tissue Tek Prisma (Sakura Finetech)
-1. Drying: 1 minute x 1
-2. Xylene: 5 minutes x 1
Xylene: 3 minutes x 3
-3. 100% alcohol (dry sol): 3 minutes x 1
100% alcohol (dry sol): 2 minutes x 2
-4. Flushing (tap water): 1 minute x 1
-5. Finish (in distilled water)
2. HE staining (GM hematoxylin: Muto Chemical, pure eosin: Muto Chemical, eosin dilution: 95% alcohol)
Uses Tissue Tek Prisma (Sakura Finetech)
-1. Flushing (tap water): 30 seconds
-2. GM hematoxylin solution: 1 minute
-3. Water washing (tap water): 30 seconds
-4. Hot water: 1 minute
-5. Flushing (tap water): 3 minutes
-6. 100% alcohol :: 15 seconds x 2
-7. Eosin solution (diluted 8 times): 30 seconds
-8. 100% alcohol :: 15 seconds x 2
100% alcohol :: 30 seconds x 1
100% alcohol :: 1 minute 30 seconds x 1
100% alcohol :: 2 minutes x 1
-9. Xylene :: 2 minutes x 2
Xylene :: 3 minutes x 1
3. IHC
(Roche's automatic immunostaining device Ventana BenchMark XT) used
-1. Slide (EZPrep) 75 ℃: 4 minutes
-2. Rinse (EZPrep) x 2
-3. Slide (EZPrep) 76 ℃: 4 minutes
-4. Rinse (EZPrep) x 1
-5. Return slide to room temperature
-6. Conditioning (Cell Conditioner Medium): 8 minutes
-7. Slide (Cell Conditioner Medium) 95 ℃: 8 minutes
-8. Conditioning (Cell Conditioner Medium): 30 minutes
-9. Slide (in Cell Conditioner Medium) 100 ℃: 4 minutes
-10. Conditioning (in Cell Conditioner Medium): 60 minutes
-11 Return slide to room temperature
-12. Rinse (Reaction Buffer) x 2
-13. Slide (Reaction Buffer) 37 ℃: 4 minutes
-14. Rinse (Reaction Buffer) x 1
-15. Slide (I-VIEW INHIBITOR) (37 ℃): 4 minutes
-13. Rinse (Reaction Buffer) x 1
-14. Slide (Reaction Buffer) 37 ℃: 4 minutes
-15. Rinse (Reaction Buffer) x 1
-16. Slide (primary antibody) 37 ℃: 32 minutes
-17. Rinse (Reaction Buffer) x 1
-18 slide (Reaction Buffer) 37 ℃: 4 minutes
-19. Rinse (Reaction Buffer) x 1
-20. Slide (I-VIEW BIOTIN Ig) (37 ℃): 8 minutes
-21. Rinse (Reaction Buffer) x 1
-22. Slide (I-VIEW SA-HRP) (37 ℃): 8 minutes
-23. Rinse (Reaction Buffer) x 2
-24. Slide (I-VIEW DAB and I-VIEW H2O2) (37 ℃): 8 minutes
-25. Rinse (Reaction Buffer) x 1
-26. Slide (I-VIEW COPPER) (37 ℃): 4 minutes
-27. Rinse (Reaction Buffer) x 1
-28. Slide (HEMATOXYLIN II) (37 ℃): 16 minutes
-29. Rinse (Reaction Buffer) x 2
-30. Slide (BLUING REAGENT) (37 ℃): 4 minutes
-31. Rinse (Reaction Buffer) x 2
The 1-fold primary antibody concentration was:

CK5 / 6 antibody (DAKO, clone: D5 / 16 B4) mouse monoclonal: 900 μg / L
calponin antibody (DAKO, clone: CALP) mouse monoclonal: 860 μg / L
Anti-Estrogen receptorα antibody (Spring Bioscience, clone: SP1) rabbit monoclonal: 200-fold dilution Anti-Progesteron antibody (Spring Bioscience, clone: SP2) rabbit monoclonal: 400-fold dilution
CK14 antibody (Novocastra, clone: NCL-L-LL002) Mouse monoclonal: 2.1 mg / L
FIG. 1 shows HE staining in non-invasive ductal carcinoma in situ (DCIS) and non-invasive lobular carcinoma (LIS) cases. DCIS is indicated by a solid line, LCIS is indicated by a broken line, and normal ductal hyperplasia (UDH) is indicated by a dotted line. The DCIS component was difficult to distinguish from papilloma. The presence or absence of myoepithelial cells was unknown. The presence of myoepithelial cells in the LCIS component was unknown. It was difficult to distinguish the UDH component from tumor epithelium. As shown by the arrows, the blood vessel component was visible.

  FIG. 2 shows hematoxylin staining + (CK5 / 6 1x) in breast DCIS and LCIS cases. DCIS is indicated by a solid line, LCIS is indicated by a broken line, and normal epithelial hyperplasia (UDH) is indicated by a dotted line. The DCIS component was slightly visible. It was difficult to determine the presence of myoepithelial cells. LCIS component was slightly visible. The presence or absence of myoepithelial cells was difficult to determine. The UDH component was confirmed. As indicated by the arrows, it was difficult to visually recognize the blood vessel component.

  FIG. 3 shows hematoxylin staining + (calponin 1x) in breast DCIS and LCIS cases. DCIS is indicated by a solid line, LCIS is indicated by a broken line, and normal epithelial hyperplasia (UDH) is indicated by a dotted line. The DCIS component was visible. The presence or absence of myoepithelial cells was easily determined. LCIS component is visible. The presence or absence of myoepithelial cells was easily determined. The UDH component was not visible. As indicated by the arrow, the blood vessel component was visible.

  FIG. 4 shows hematoxylin staining + (CK5 / 6 1x) + (calponin 1x) in breast DCIS and LCIS cases. DCIS is indicated by a solid line, LCIS is indicated by a broken line, and normal epithelial hyperplasia (UDH) is indicated by a dotted line. The DCIS component was visible. The presence or absence of myoepithelial cells was easily determined. The LCIS component was visible. The presence or absence of myoepithelial cells was easily determined. The UDH component was visible. As indicated by the arrow, the blood vessel component was visible.

  FIG. 5 shows hematoxylin staining + (CK5 / 6 1 / 2x) + (calponin 1 / 2x) in breast DCIS and LCIS cases. DCIS is indicated by a solid line, LCIS is indicated by a broken line, and normal epithelial hyperplasia (UDH) is indicated by a dotted line. The DCIS component was visible. The presence or absence of myoepithelial cells was easily determined. The LCIS component was visible. The presence or absence of myoepithelial cells was easily determined. The UDH component was visible. As indicated by the arrows, the blood vessel component was visible.

  FIG. 6 shows hematoxylin staining + (CK5 / 6 1 / 4x) + (calponin 1 / 4x) in breast DCIS and LCIS cases. DCIS is indicated by a solid line, LCIS is indicated by a broken line, and normal epithelial hyperplasia (UDH) is indicated by a dotted line. The DCIS component was visible. The presence or absence of myoepithelial cells was easily determined. The LCIS component was visible. The presence or absence of myoepithelial cells was easily determined. The UDH component was visible. As indicated by the arrows, the blood vessel component was visible.

  FIG. 7 shows hematoxylin HE staining + (CK5 / 6 1 / 8x) + (calponin 1 / 8x) in breast DCIS and LCIS cases. DCIS is indicated by a solid line, LCIS is indicated by a broken line, and normal epithelial hyperplasia (UDH) is indicated by a dotted line. The DCIS component was visible. The presence or absence of myoepithelial cells was easily determined. The LCIS component was visible. The presence or absence of myoepithelial cells was easily determined. The UDH component was visible. As indicated by the arrows, the blood vessel component was visible.

  FIG. 8 shows hematoxylin staining + (ER 1x) in breast DCIS and LCIS cases. DCIS is indicated by a solid line, LCIS is indicated by a broken line, and normal epithelial hyperplasia (UDH) is indicated by a dotted line. The DCIS component was visible. The presence or absence of myoepithelial cells was difficult to determine. The LCIS component was visible. The presence or absence of myoepithelial cells was difficult to determine. The UDH component was visible. As indicated by the arrows, it was impossible to visually recognize the blood vessel component.

  FIG. 9 shows hematoxylin staining + (PgR 1x) in breast DCIS and LCIS cases. DCIS is indicated by a solid line, LCIS is indicated by a broken line, and normal epithelial hyperplasia (UDH) is indicated by a dotted line. The DCIS component was difficult to see. The presence or absence of myoepithelial cells was difficult to determine. The LCIS component was difficult to see. The presence or absence of myoepithelial cells was difficult to determine. The UDH component was visible. As indicated by the arrows, the vascular component was not visible.

  FIG. 10 shows hematoxylin staining + (ER 1x) + (PgR 1x) in breast DCIS and LCIS cases. DCIS is indicated by a solid line, LCIS is indicated by a broken line, and normal epithelial hyperplasia (UDH) is indicated by a dotted line. The DCIS component was visible. The presence or absence of myoepithelial cells was difficult to determine. The LCIS component was visible. The presence or absence of myoepithelial cells was difficult to determine. The UDH component was visible. As indicated by the arrows, the blood vessel component was not visible.

  FIG. 11 shows hematoxylin staining + (ER 1 / 2x) + (PgR 1 / 2x) in breast DCIS and LCIS cases. DCIS is indicated by a solid line, LCIS is indicated by a broken line, and normal epithelial hyperplasia (UDH) is indicated by a dotted line. The DCIS component was visible. The presence or absence of myoepithelial cells was difficult to determine. The LCIS component was visible. The presence or absence of myoepithelial cells was difficult to determine. The UDH component was visible. As indicated by the arrows, the blood vessel component was not visible.

  FIG. 12 shows hematoxylin staining + (ER 1 / 4x) + (PgR 1 / 4x) in breast DCIS and LCIS cases. DCIS is indicated by a solid line, LCIS is indicated by a broken line, and normal epithelial hyperplasia (UDH) is indicated by a dotted line. The DCIS component was visible. The presence or absence of myoepithelial cells was difficult to determine. The LCIS component was visible. The presence or absence of myoepithelial cells was difficult to determine. The UDH component was visible. As indicated by the arrows, the blood vessel component was not visible.

  FIG. 13 shows hematoxylin staining + (ER 1 / 8x) + (PgR 1 / 8x) in breast DCIS and LCIS cases. DCIS is indicated by a solid line, LCIS is indicated by a broken line, and normal epithelial hyperplasia (UDH) is indicated by a dotted line. The DCIS component was difficult to see. The presence or absence of myoepithelial cells was difficult to determine. The LCIS component was difficult to see. The presence or absence of myoepithelial cells was difficult to determine. The UDH component was visible. As indicated by the arrows, the blood vessel component was not visible.

  FIG. 14 shows hematoxylin staining + (ER 1 / 16x) + (PgR 1 / 16x) in breast DCIS and LCIS cases. DCIS is indicated by a solid line, LCIS is indicated by a broken line, and normal epithelial hyperplasia (UDH) is indicated by a dotted line. The DCIS component was difficult to see. The presence or absence of myoepithelial cells was difficult to determine. The LCIS component was difficult to see. The presence or absence of myoepithelial cells was difficult to determine. The UDH component was not visible. As indicated by the arrows, the blood vessel component was not visible.

  FIG. 15 shows hematoxylin staining + (ER 1x) + (PgR 1x) + (calponin 1x) in breast DCIS and LCIS cases. DCIS is indicated by a solid line, LCIS is indicated by a broken line, and normal epithelial hyperplasia (UDH) is indicated by a dotted line. The DCIS component was visible. The presence or absence of myoepithelial cells was easy to determine. The LCIS component was visible. The presence or absence of myoepithelial cells was easy to determine. The UDH component was difficult to see. As indicated by the arrows, the blood vessel component was visible.

  FIG. 16 shows hematoxylin staining + (ER 1x) + (PgR 1x) + (calponin 1 / 2x) in breast DCIS and LCIS cases. DCIS is indicated by a solid line, LCIS is indicated by a broken line, and normal epithelial hyperplasia (UDH) is indicated by a dotted line. The DCIS component was visible. The presence or absence of myoepithelial cells was easy to determine. The LCIS component was visible. The presence or absence of myoepithelial cells was easy to determine. The UDH component was visible. As indicated by the arrows, the blood vessel component was visible.

  FIG. 17 shows hematoxylin staining + (ER 1x) + (PgR 1x) + (calponin 1 / 4x) in breast DCIS and LCIS cases. DCIS is indicated by a solid line, LCIS is indicated by a broken line, and normal epithelial hyperplasia (UDH) is indicated by a dotted line. The DCIS component was visible. The presence or absence of myoepithelial cells was easy to determine. The LCIS component was visible. The presence or absence of myoepithelial cells was easy to determine. The UDH component was visible. As indicated by the arrow, the blood vessel component was visible.

  FIG. 18 shows hematoxylin staining + (CK14 1 / 2x) + (calponin 1 / 2x) in breast DCIS and LCIS cases. DCIS is indicated by a solid line, LCIS is indicated by a broken line, and normal epithelial hyperplasia (UDH) is indicated by a dotted line. The DCIS component was visible. The presence or absence of myoepithelial cells was easily determined. The LCIS component was visible. The presence or absence of myoepithelial cells was easily determined. The UDH component was visible. As indicated by the arrows, the blood vessel component was visible.

  FIG. 19 shows hematoxylin staining + (CK14 1 / 4x) + (calponin 1 / 4x) in breast DCIS and LCIS cases. DCIS is indicated by a solid line, LCIS is indicated by a broken line, and normal epithelial hyperplasia (UDH) is indicated by a dotted line. The DCIS component was visible. The presence or absence of myoepithelial cells was easily determined. The LCIS component was visible. The presence or absence of myoepithelial cells was easily determined. The UDH component was visible. As indicated by the arrows, the blood vessel component was visible.

  It can be used for pathological diagnosis.

Claims (5)

A cocktail antibody comprising an anti-estrogen receptor antibody, an anti-progesterone antibody and an anti-calponin antibody, which stains a single pathological specimen material section that is physically identical. Anti-cytokeratin 5/6 antibody or anti-cytokeratin 5 antibody or anti-cytokeratin 14 antibody or anti-cytokeratin 5/14 antibody or anti-high molecular weight cytokeratin that stains a single pathological specimen material that is physically identical A cocktail antibody comprising an antibody (34βE12 antibody) and an anti-calponin antibody. The concentration of the anti-cytokeratin 5/6 antibody is 0.9 μg / L to 45 mg / L, the concentration of the anti-calponin antibody is 0.86 μg / L to 86 mg / L, and the concentration of the anti-cytokeratin 14 antibody is 2.1 The cocktail antibody according to claim 2 , wherein the cocktail antibody is μg / L to 42 mg / L. The cocktail antibody according to any one of claims 1 to 3 , wherein the pathological specimen material section is a mammary neoplastic section. The cocktail antibody according to any one of claims 1 to 3 , wherein the pathological specimen material section is a mammary gland non-neoplastic section.