RU2417056C1 - Diagnostic technique for malignant process in human body - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: invention refers to medicine, namely to diagnosing malignant processes in a human body. A diagnostic technique for a malignant process in a human body consisting in sampling a patient's living tissue, grinding, mixing with physiologic saline to a state of suspension, keeping and agitating under certain conditions, then centrifuging, separating supernatant and detecting cancer-specific markers by an immunochemiluminisent assay.

EFFECT: method exhibits a simple and high degree of malignant process detection in the human body, both at the early, and following stages of disease.

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Description

The invention relates to medicine, namely to the diagnosis of malignant processes in the human body. Oncological diseases are one of the most common causes of death, and often at a young age. Successful treatment is possible only in the early stages, which requires their early detection. The identification of tumor markers allows you to successfully prevent and treat this severe pathology. Oncomarkers are specific substances that are formed as a result of the activity of cancer (and sometimes normal) cells and are usually found in blood and / or urine in cancer patients. They are extremely diverse in structure, although in most cases they are proteins or their derivatives. Some of them are highly specific, i.e. characteristic of one type of tumor, but at the same time can be detected in other forms of cancer. A targeted search for specific and cheap tumor markers is underway. The new tumor markers were given numbered names, for example, CA 19-9 for cancer of the rectum and pancreas, CA 15-3 for breast cancer and CA 125 for ovarian cancer. Many other open markers for the diagnosis of these types of cancer have not been further studied because they have not shown their advantage over the above. But, unfortunately, even the specificity and sensitivity of recognized tumor markers leaves much to be desired. Firstly, in almost every person a small number of tumor markers can be detected in the blood, which complicates the diagnosis at an early stage of tumor development. A significant increase in the tumor marker level is often observed at later stages, when the tumor is already sufficiently developed. Secondly, even with a sufficiently high level of tumor markers in the blood, many of them are nonspecific: CEA, originally discovered as a marker of colon cancer, can be increased in patients with lung and breast cancer, and a high level of CA 125 is found in women with gynecological non-tumorous diseases. Despite the fact that tumor markers were developed specifically for the early (even at an asymptomatic stage) cancer diagnosis, in practice, only the prostate-specific antigen (PSA) is appropriate. In all other cases, a reliable diagnosis of cancer is established only by the results of studying under the microscope a biopsy (piece of tissue) of the tumor. The main use of tumor markers in clinical diagnosis is monitoring the course of the disease and the effectiveness of the treatment, obtaining prognostic information. One of the problems solved by using tumor markers is the possibility, in combination with other diagnostic methods, of early differential diagnosis of the tumor and monitoring the effectiveness of treatment. The dynamics of an increase in the level of a tumor marker usually allows us to make a conclusion about the presence and nature of the development of the disease and metastasis and is of more interest than a single value of the level taken by itself. With regular monitoring of the level of markers that are informative for a tumor of a specific location, metastases can be detected 4-6 months before their clinical detection. During treatment or after surgery, the absence of a decrease in the concentration of tumor markers means the inefficiency of the selected treatment method. Unfortunately, no tumor marker possessing 100% specificity with respect to any organ has yet to be characterized.

Closest to the claimed is a method of morphological diagnosis of malignant processes, which consists in extracting a patient’s tissue sample, maintaining the tissue in a fixed state using fixatives, dehydration and further paraffin filling. After pouring with paraffin and solidification of the sample, the preparation is made using a microtome. Then sections from a paraffin sample are placed on a glass slide, paraffin is removed, stained and covered with a coverslip. The prepared preparation is placed under a microscope to determine the ultrastructure of cells and their components and to diagnose malignant processes (V. Eliseev and others. 1972).

The disadvantage of this method is the complexity of the process of preparing a diagnostic drug, as well as the high probability of obtaining an unreliable result, since the diagnosis does not examine the entire volume of tissue extracted from the patient’s organ, but only sections, which complicates the diagnosis in the early stages of tumor development.

Closest to the claimed is a device for diagnosing malignant processes, which contains a sequentially connected fixation unit, a dehydration unit, a thermostat, a microtome, a staining unit, a microscope (V. Eliseev and others. 1972).

A disadvantage of the known device is the complexity of the process of preparing a diagnostic drug, as well as manual labor at some of its stages, which reduces the quality of the resulting drug and the reliability of the diagnosis of malignant processes in the human body.

The technical result, which is achieved in the claimed method for diagnosing a malignant process, is the simplicity and high probability of detecting malignant processes in the human body both in the early stages of the disease and in the later ones. This is achieved due to the fact that by the level of the tumor marker it is possible to judge the presence of deviations not only in the area from which the tissue sample was taken, but also about the vast distribution of the oncological process.

The technical result is achieved by the fact that in the method for diagnosing a malignant process in a human body, the patient’s living tissue is taken, crushed, mixed with 2 ml of physiological saline to a suspension state, then incubated for 5 minutes at a temperature of 20 ° C, then mixed and again incubated for 5 minutes at at a temperature of 20 ° C, while the steps of maintaining and mixing the suspension are repeated 3 times, then centrifuged at 10,000 rpm for 2 minutes, the supernatant is selected and the tumor mark is determined by immunochemiluminescent method ry.

The method is implemented as follows.

A tissue sample of the patient is placed in a test tube and crushed using a sterile tip or a special pestle. To the crushed tissue, add 2 ml of physiological saline and mix on a vortex until a suspension is obtained. Then the tube is placed in a thermostat at 20 ° C for 5 minutes. Remove the tube from the refrigerator, mix it on a vortex and place it in the thermostat again. The cooling, stirring and cooling procedures are repeated 3 times. The tissue tube in physiological saline is then centrifuged at 10,000 rpm for 2 minutes. The supernatant is selected in a clean tube, in which the tumor marker is determined using the standard immunochemiluminescent method. The table shows the data oncomarker levels for diagnosis by blood and by the present method.

Marker levels in blood and tumor tissue in cancer patients depending on the stage of the disease Patient No. ist. / Illness Disease diagnosis Blood marker Tumor Marker Level B-naya A. No. 23 Breast Cancer T1NoMo CA15-3: 0.10 U / ml CA15-3: 73.82 U / ml B-naya R. №43 Breast Cancer T1N1Mo CA15-3: 6.10 U / ml CA15-3: 123.8 U / ml B-naya K. №48 Breast Cancer T3N2M1 CA15-3: 46.1 U / ml CA15-3:> 200 U / ml B-NoA. Number 13 Gastric Cancer T1NoMo CEA: 0.74 ng / ml CEA: 35.7 ng / ml B-no. Number 63 Gastric Cancer T2N1Mo CEA: 3.74 ng / ml CEA: 43.74 ng / ml B-naya K. No. 73 Gastric Cancer T3N1M1 CEA: 23.74 ng / ml CEA: 146.74 ng / ml B-naya K. №48 Uterine Cancer T3N1M1 CEA: 23.74 ng / ml CEA: 223.84 ng / ml B-naya K. №48 Ovarian Cancer T1NoMo CA125: 2.0 U / ml CA 125: 56.10 U / ml B-naya K. №48 Ovarian Cancer T3N2M1 CA 125: 196.10 U / ml CA 125:> 200 U / ml

Normal content in blood serum: CA15-3 and CA125 - less than 30 U / ml; CEA - 10 ng / ml.

The table shows that the diagnosis of the disease by the level of tumor marker in the blood has a very low percentage of detection of diseases (actually at the stage of the presence of metastases).

The claimed method allows to identify diseases at earlier stages, in which treatment is more effective and, depending on the level of the marker, correct the stage of the disease.

The inventive method for diagnosing a malignant process in the human body is a high-tech, simple diagnostic test, which allows a high degree of certainty to identify malignant processes in the human body, both in the early and later stages of the disease and to correct treatment.

Claims (1)

A method for diagnosing a malignant process in the human body, namely, that they take live tissue from a patient, grind it, mix it in 2 ml of physiological saline to a suspension state, then hold it for 5 minutes at a temperature of 20 ° C, then mix it and again hold it for 5 minutes at a temperature 20 ° C, while the aging and stirring steps are repeated 3 times, then centrifuged at 10,000 rpm for 2 minutes, the supernatant is selected and the tumor marker is determined by the immunochemiluminescent method.