Patent RU-2677872-C1 (RU)
Inventor: Vasilev Nikolaj Voltovich
IPC: G01N33/48
Filed: 2017-08-01
Granted: 2019-01-22
Abstract: FIELD: medicine.SUBSTANCE: invention relates to the field of medicine, specifically to oncology, and can be used to predict lymphangitic metastasis in malignant tumors from the tunics of peripheral nerves. Essence of the method: conduct a macroscopic study of the primary tumor, tissue of the resection lines and all lymph nodes removed during the operation and morphological study of the preparations of the tissue of the primary tumor at the light-optical level, the presence of atypical mitoses figures in tumor cells is evaluated, for this purpose, in 10 fields of view, selected at random, when increasing the lens of the microscope ×400 consider the number of figures of atypical mitoses, in the absence of mitoses, this sign is estimated at 1 point; if there are single mitoses, up to 5 mitoses in 10 fields of view, a sign is estimated at 2 points, in the presence of more than 5 atypical mitoses, the sign is estimated at 3 points, then in the tumor tissue, a morphological assessment of spontaneous necrosis is carried out with determination of the area of spontaneous necrosis in the primary tumor within the histological specimen with an increase in the objective of the microscope ×100, in the absence of spontaneous necrosis in the tumor tissue, the symptom is estimated at 1 point, if the necrosis zone occupies less than 10 % of the tumor area within the histological section, the symptom is evaluated at 2 points, in cases when the area of spontaneous necrosis in the tumor tissue takes from 10 to 50 % of the area – a sign is estimated at 3 points, if more than 50 %, a sign is estimated at 4 points, after a histological examination of the tissue of the primary tumor, an immunohistochemical study is performed using antibodies to SMA and S100 protein (Dako) according to standard procedures according to protocols, then the expression of these markers in tumor preparations is evaluated, in the presence of negative expression of the SMA marker, a sign is estimated at 1 point, with moderate positive expression, the sign is estimated at 2 points, with mild positive expression, the sign is estimated at 3 points, in the presence of negative expression of the S100 marker, the sign is estimated at 1 point, with moderate positive expression, the sign is estimated at 2 points, with mild positive expression, the sign is estimated at 3 points, in addition, in each case, an immunohistochemical study of a wide range of cytokeratins is performed in the tumor tissue, and in each case 11 markers are studied using antibodies: Pan-Cytokeratin (5/6/8/18) (Clone 5D3, Novocastra), Cytokeratin AE1/AE3 (Clone AE1/AE3, Dako), Cytokeratin 5 (Clone XM26, Novocastra), Cytokeratin 5/6 (Clone D5/16 B4, Dako), Cytokeratin 7 (Clone OV-TL 12/30, Novocastra), Cytokeratin 8 (Clone TS1, Novocastra), Cytokeratin 14 (Clone LL002, Novocastra), Cytokeratin 19 (Clone b170, Novocastra), Cytokeratin 20 (Clone K20.08, Novocastra), Cytokeratin HMW (betta Clone 34E12, Dako), Cytokeratin 8/18 (Clone 5D3, Dako), expression of each marker is assessed as negative or positive, with negative expression of all the above markers, this characteristic is estimated at 1 point, in the presence of positive expression of at least one of 11 cytokeratins – the value of the regression equation of the listed markers in the tumor tissue – the sign is estimated at 2 points, further, the value of the regression equation Y is calculated by the formula, which is used in the formula to determine the value of the probability of development of lymphangitic metastases P. With P≥50 % define high, and with P<50 % – low risk of lymphangitic metastasis. Sensitivity of the method was 100 %, the specificity of 100 %.EFFECT: invention provides improved accuracy and information value of method.1 cl, 2 ex