RU2285443C2 - Method for stabilizing individual substrate for spectral malignance determination - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: method involves using individual substrate having initial composition as follows. 93.8-98.5% by mass take pleural exudates or pericardial exudates or ascites, 1.5-6.2% by mass of stabilizer for separating nebulous and transparent centrifugate, 10.0-26.8% by mass of transparent diluter like ethanol, dioxane or tetrahydrofuran of spectral purity. The exudates composed of pleural exudates or pericardial exudates or ascites are taken in usual sterile way from patient organism in the amount of 93.8-98.5% by mass. Stabilizer is added as 10% of aqueous solution of guanidine sulfate, cesium sulfate, primary potassium biphosphate or ammonium sulfate in the amount of 1.5-6.2% by mass. The obtained mixture is centrifuged at 22.8-26.6 rpm during 800-1050 s. Aliquot part is taken from the whole volume of produced transparent sample equal to 74.5-90.0% by mass and 10.0-26.8% by mass are of transparent solvent. Individual substrate sample obtained in this way is frozen at 248-257 K before being used or it is used immediately in carrying out spectral analysis for determining malignance.

EFFECT: high accuracy of diagnosis.

3 dwg, 1 tbl

Description

FIELD OF THE INVENTION

The invention relates to a new method for stabilizing an individual substrate for spectral determination of malignancy in medicine.

Technical problem

The technical problem solved by the present invention is the implementation of a new method of stabilization of an individual substrate for spectral determination of malignancy in medicine. This allows you to use the well-known UV-VIS diagnostic methods for spectral screening as follows: the method is carried out in vitro (after extraction of body fluids) and regardless of the place of extraction at any time and anywhere where it is possible to carry out.

State of the art

Diagnosing is a common clinical and technical problem. Any additional information in the diagnosis, in the sense of additional diagnostic methods, is very important because it complements the picture of the pathological process in the body and in this way facilitates the establishment of the correct final diagnosis.

The main problem of each diagnostic method (classic and additional) is that it should be the most possible less invasive or non-invasive, fast and reliable. An example of the difficulty of such a task is the diagnosis of hypertension, i.e. the most accurate blood pressure measurement is performed by an invasive method using an intra-arterial catheter. Due to invasiveness, technical limitations and financial reasons, this method is not suitable for daily use and screening (Momčilo Babic, Skrining u medicini, Markom-publik, Beograd, 2001).

Unfortunately, the accuracy of a diagnostic method is often proportional to its invasiveness, and therefore it is used as the last diagnostic method. Therefore, work is underway to develop and discover additional diagnostic methods that are less invasive, but usually less accurate.

Given that all methods have their advantages and disadvantages and that none of them is 100% reliable or one-way, the final diagnosis is always established using several different diagnostic methods. The solution to the problem of correct diagnosis is achieved by the development of new additional, quick, screening diagnostic methods that facilitate and accelerate the usual standard clinical procedure.

Numerous diagnostic methods are known in oncology and pulmonology, for example, X-ray analysis, ultrasound, nuclear magnetic resonance, CT scan MRI (CT NMR slice) pleurocentesis, diseased tissue biopsy, blood tests, etc. (Ruckdeschel JC. Malignant Pleural Effusions, Recent Advances in Diagnosis and Management, Princeton, J. Jersey, Bristol Myers Squibb Company, 1992, 8-11). The number and presence of malignant cells, for example, in pleural effusion cannot be determined by cytological analysis in only one trial. Several consecutive tests give a more reliable result, but not always. In the case of a tissue biopsy, the tissue is a substrate for a further diagnostic method. The analyzed tissue should be representative, i.e. must satisfy certain conditions, namely, the sample must contain the tissue of the test tumor, as well as part of the tissue bordering normal tissue. It is also important that there are no secondary changes in the tumor tissue (necrosis, bleeding, etc.). The largest number of false positive and false negative diagnoses was established specifically on tissue samples that were squeezed, coagulated, swollen from blood, necrotic, fixed inappropriately or due to insufficient material.

Currently, the following diagnostic methods are used in the diagnosis of cancer:

1. Biopsy ex tempore (simultaneous) (Frozen Section, FS).

2. Aspiration biopsy with cytological smear (Fine needle Aspiration, FNA).

Also used:

3. Histochemistry.

4. Immunohistochemistry.

5. Electron microscopy.

6. Tissue culture (in situ hybridization, receptor autoradiography and other new methods that are not available to us for financial reasons).

Ex tempore biopsy must be accurate, reliable and fast, and in order to do this, there must be appropriate preconditions for work:

1. Good knowledge of pathology (solid experience).

2. Representative material (well prepared substrate).

3. Relevant specifications.

If the above conditions are met, an experienced pathologist can make an accurate diagnosis in 90-96% of cases, taking into account long-term responses (with long-term medications), which is essential, since an accurate diagnosis allows the surgeon to immediately apply a very long surgical intervention.

Representative material implies that the pathologist will receive enough tissue and that the sample contains precisely the pathological change with part of the tissue surrounding it. When performing an ex tempore biopsy in cases in which malignancy is suspected, there are three possible answers:

(a) positive for malignancy,

(b) negative for malignancy,

(c) the diagnosis is delayed.

(Beahrs OH: Staging of cancer of the breast as a guide to therapy. Cancer 53, 592, 1984; Bugis SP, Yuoung JEM et al .: Diagnosis accuracy of fine needle aspiration biopsy versus frozen section in solitary thyroid nodules. Amer J Surg 152, 411, 1986; Carter D .: Interpretation of Breast Biopsies. New York, Raven Press, 1988.)

Cytological smears (Pap tests) provide another method for diagnosing cancer. This method is widely used in the determination of cervical cancer, often in the pre-invasive stage, but is also used to confirm cases of potential malignancy, such as endometrial cancer, bronchogenic cancer, bladder and prostate cancer, and stomach cancer. This method is also used to identify tumor cells in abdominal, pleural, synovial and cerebrospinal fluids.

As you know, cancer cells lose their adhesion, peel off and fall off, and show the whole spectrum of morphological changes covered by the term anaplasia. Therefore, flake cells should be tested for signs of anaplasia indicating their origin. In contrast to histological analysis, the findings are based solely on a violation of the cytology of an individual cell or sometimes a small group of cells, without signs of architectural disruption, loss of orientation, or evidence of invasion. This method allows us to differentiate between normal, dysplasic, and cancer cells, and sometimes provides for the detection of cellular changes characteristic of in situ carcinoma.

Smears of other types of tumors - lungs, stomach, prostate - are much more difficult to evaluate. It should be emphasized that all positive results must be confirmed by biopsy and histological testing, before using any therapy. It should also be pointed out that a negative cytological result does not exclude a malignant tumor.

Although histology and exfoliative cytology remain the methods most often used in the diagnosis of cancer, new methods are constantly appearing that contribute to a faster and more accurate diagnosis. (Cancer statistics, 1989, Cancer 39, 13, 1989; Robbins L.S., Cotran R.S., Kumar V .: Pathologic Basis of Disease, 4th ed. WB Saunders Co. Philadelphis 1989.)

In the case of a malignant process in the lungs, pleuroscopy is the most reliable method, but as with the diagnosis of hypertension by an intra-arterial catheter, it is the most aggressive and invasive method for humans. Laparoscopy uses the so-called laparoscopy of media stinuma and the pleural cavity, known as the invasive technique. When studying literature, one can find data on the development of new, additional diagnostic methods that are either less invasive than the above or more reliable than cytological analysis in the sense of establishing a qualitative final diagnosis.

A detailed examination of the available scientific and specialized literature revealed that there are very few references related to this problem, while at the same time trying to overcome the shortcomings of the known invasive diagnostic methods. For example, "Biomax Technologies Inc." in its patent application of June 24, 1999 (WO 9966830, CA 9900586) describes a device and methods that allow direct examination of tissue induced by fluorescence using an endoscope, without the use of large and expensive auxiliary devices. On the other hand, Board of Regents, The University of Texas System, in its application of 12/24/1997 (WO 97/48329, PCT / US 97/10204) describes Raman NIR spectroscopy for in vitro and in vivo determination of precancerous lesions of the cervix. None of these patent applications in their works describes such a simple method of early diagnosis as our method provides using relatively simple UV-VIS spectroscopy, indicating the presence of a malignant tumor in a stabilized substrate in vitro.

SUMMARY OF THE INVENTION

The method of stabilizing an individual substrate in vitro for spectral determination of malignancy includes, in accordance with the present invention, adding 10 wt.% Aqueous solutions of stabilizers to human body fluid taken in vivo (pleural punctate, ascites, pericardial effusion). The resulting mixture was centrifuged. An aliquot of the total volume of the obtained transparent sample is taken. Then add transparent solvents of spectral purity. The resulting sample of an individual substrate is frozen at 248-257 K before use or the spectrum is immediately taken on a UV-VIS spectroscope.

According to a preferred embodiment of the invention, stabilizers having the function of denaturing and stabilizing body fluids, which, among other things, contain proteins, are ammonium sulfate, cesium sulfate, potassium primary phosphate and guanidinium sulfate.

According to a most preferred embodiment of the invention, aqueous solutions of ammonium sulfate or potassium hydrogen phosphate are used as stabilizer. At the same time, in accordance with the most preferred embodiment of the present invention, 95% ethanol and tetrahydrofuran are transparent solvents of spectral purity.

All UV-VIS spectra were recorded at a wavelength of 200-800 nm. Two types of spectra were obtained, one characteristic of malignant diseases, and the second for benign diseases, the presence of which is uniquely detected in pleural effusions, ascites, and pericardial effusions.

In the spectrum of the first type at 230-275 nm, two peaks are detected. Both peaks are very well designed and defined. The absorption of both peaks has a value between 4 and 5. In the range from 380 to 500 nm, one very flat plateau with a low absorption value (about 0.5) was found. This is the usual type of spectrum of all these body fluids (pleural effusions, ascites and pericardial effusions) in the presence of a malignant disease in any part of the body. In 620 of 650 tested body fluids with suspected malignancy in the body, this type of spectrum was obtained. The diagnosis made by UV-VIS spectroscopy was confirmed and coincides with the diagnosis made by the usual clinical, radiological, cytological and histopathological procedures.

The second type of obtained spectra showed instead of two well-defined peaks in the region of 230-280 nm one very wide band, which at the top mainly has a lot of noise. The absorption value is 4-5. This broad band is typical for body fluids resulting from benign diseases in the body. In the range from 380 to 599 nm, a high plateau appears, which is four to five times higher than the plateau in the case of malignant diseases. A diagnosis of the absence of tumor malignancy was confirmed in 300 of 325 tested body fluids (pleural effusion, ascites, pericardial effusion). A significant difference is that a new screening diagnostic method can detect a malignant disease in the three tested types of body fluids.

The table contains all the data in connection with the selected sample of a certain number (30) of patients whose stabilized body fluid was analyzed, in which the diagnosis made by UV-VIS spectroscopy unambiguously coincides with the clinical diagnoses established by various clinical methods.

Patient Code Age / gender Exudate (type of taken body fluid) Clinical diagnosis Cytology result UV-VIS diagnosis XX02 1967 / f pericardial effusion Non-Hodgkin lymphoma st IVB; St. post irrad. CNS am I / iand VCS ami malignant cells M Xx05 1914 / f pleural effusion Adenocarcinoma pulm.I. dex. M Xx07 1951 / f ascites St.post.resec.sigmae pp ca aa II M Xx08 1939 / f ascites St.post.amp. Mammae I.sin pp Ca aaXII; Effusio pleurae I.sin. maligna; Pleurodesis am III; Ascites zlokach. cells originating from the site of carcinoma M Xx09 1947 / f ascites St.post hysterectomiam totalis cum adnexextomiam bill.pp ca ovarii st.IC M XY10 1934 / m ascites Neo ventriculi cum meta hepatitis M XX13 1940 / w ascites Ca ventriculi typus diffusum (anaplasticum) cum meta in lymphonodis zlokach. cells derived from adenocarcinoma M Xx14 1948 / f ascites St.post.amput. Mammae I.dex.pp Ca. Meta hepatis M Xx22 1925 / w ascites Tu ovarii lat dex. M XY24 1945 / m ascites Adenocarcinoma partim anaplasticum f. ventriculi. Meta hepatitis. Carcinosis peritonei. Ascites M XY27 1941 / m ascites Ascites. Carcinosis peritonei ex origine ignote. adenocarcinoma cells, part of a giant cell M Xx28 1940 / w pleural effusion St.post.amp mammae I. dex. pp Ca ductale inv. exilic. Gr. Ill aa III. St.post. irrad. Post op.Meta. Pulmo. bil. cum pleuritis carcinomatosa I. dex. Meta ossium gen. St.post. incultus vascularis cerebri. M Xx29 1956 / f pleural effusion St.post.amp. Mammae I. dex. pp Ca ductale inv. GIII aalX, meta hepatis, meta pulmonum. M XX30 1938 / f ascites St.post.amp. mammae I. Dexpp Ca aaXXSt. post irradiationem praeoper. ssXX Hepattomeglia. Infiltratio hepatis susp.Ascites. Effusio pleurae I. dex. Infiltratio mal. Peritonei. Cystitis ovarii I. dex. Hzdronephrosis ren. Pancreatitis chronica. Noduli erythrocytes, lymphocytes, macrophages and the mass of individual and papillary tumor cells. Malignant. origin from the outbreak weakly different. gigantokl. carcinomas M Xy32 1953 / m ascites Adenocarcinoma corporis ventriculi typus diffusum inop. carcinosis peritonei. ascites. hypoalbuminemia. St.post. lap.Expl. a.m. II. M Xx33 1945 / f ascites St.post. amp.m.Isin. p.p. Ca aa V. St. post. TCT postop. A.a. IV / V. St.post. ovariect. bil. Pp Meta aa II meta ossium gen. St.post. ira. Palitiva coxae pp meta pangastritis chronica erosiva. anemia complexa sec. trombocythopaen. M Xx34 1972 / f pericardial effusion Morbus Hodgkin. M Xx35 1920 / f ascites Tu ovarii sus. Ascites. Effusio pleurae bil. M Xx45 1952 / f pleural effusion St.post. AMLD pp. Ca ductale infiltrativum aa V / VI. Recidivum locale. Effiisio pleurae mixed cell saturation M Xx46 1936 / f ascites Ca ovarii st IV. Carec inosis perifonei parietalis et visceralis M Xx47 1931 / f ascites Laesio hepatitis. Ascites. St.post. nephrectomiam I. dex. St.post. ICV malignant meselioma M Xx50 1946 / f pleural effusion Status post laryngectomiam partialis horisontalis supragolottica et explorationem cillibilateralis A.A. III Ca laryngis recidivans M Xx51 1975 / f ascites St. post. extirpationem Tu mesosigmae a.a.I / II Ph: Schwanoma cellulare M XY54 1930 / m ascites Ca ventriculi, meta hepatis, carcinosis peritonei erythrocytes, macrophages, and meselioma cells M Xx58 1924 / w ascites Tu abdominalis M Xx59 1934 / f pleural effusion Schwanoma malignum regio cruris lat. Dex. susp. Meta M XX61 1942 / f ascites Tu ovarii sin. M Xx62 1946 / f ascites St.post. extirpatio I. gli. colli. I. sin. pp M XY65 1931 / m ascites St.post. resecsigmae. Meta hepatis M Xx66 1934 / f ascites Adenocarcinoma uteri necroticum M

The table shows the age of the patients, the type of test body fluid (exudate), clinical diagnosis and cytological result. The last column shows the result (diagnosis) obtained by the UV-VIS spectral method and the presence of a malignant tumor indicated by M. Analyzed fluids (exudates): pleural punctate (fluid that is a consequence of a pathological process in the body and accumulates in the pleural space), ascites ( fluid accumulating as a consequence of the pathological process in the peritoneal cavity) and pericardial effusion (fluid accumulating as a consequence of the pathological process between two leaves of the pericardium).

The table clearly shows (on samples selected from 30 patients) that the clinical diagnosis, i.e. the presence of a malignant tumor in various tumor processes unambiguously coincides with the diagnosis made by UV-VIS spectroscopy. Regardless of the type of process of a malignant tumor in the body (some examples from the table: ХХ05 - Adenocarcinoma pulm.I.dex., XY10 - Neo ventriculi cum meta hepatitis, XX59 - Schwanoma malignum, XX66 - Adenocarcinoma uteri necroticum, XX02 - Non-Hodgkin lymphoma) by UV-VIS spectroscopy, the presence of a malignant tumor was unambiguously detected (M is indicated in the table).

Regardless of the type of body fluid tested (exudate), the presence of a malignant process was detected by UV-VIS spectroscopy and clearly coincides with the clinical diagnosis obtained by standard clinical methods for diagnosing cancer (biopsy, histopathological examination). This is shown by the examples from the table: in the pericardial effusion (XX02), the presence of a malignant tumor - Non-Hodgkin lymphoma, indicated in Table M. The presence of a malignant tumor (XX46) - Ca ovarii st IV was detected in ascites. Carcinosis peritonei pariet alis et visceralis, is indicated in table M.

Information on the disease of all the studied patients was collected in accordance with the standard clinical protocol and form, including age, gender, profession, first symptoms, smoking habit, body weight and other tests. It was characteristic of all the studied patients with pleural or pericardial effusions that at the time the UV-VIS spectra were taken, they were not diagnosed and 90% of the patients did not undergo treatment (except for those who did not say what treatment was taken outside the medical history) control and knowledge of the doctor). So far, trials have been performed on 232 patients with pleural effusions, 565 with ascites and 178 with pericardial effusions. The average age of the tested patients is 44.6 years with a male: female ratio of 519: 456.

DETAILED DESCRIPTION OF THE INVENTION

The method of stabilizing an individual substrate in vitro for spectral determination of malignancy includes, in accordance with the present invention, adding 1.5-6.2 wt.% 10% aqueous solution of the stabilizer to in vivo human body fluids (pleural punctate, ascites, pericardial effusion) and the resulting the mixture is centrifuged for 800-1050 seconds at a speed of 22.8-26.6 rpm. A sample of the obtained clear content was taken and 10.0-26.8 wt% of a clear solvent of spectral purity was added. The resulting individual substrate sample is frozen at 248-257 K before use or the spectrum is immediately removed.

The advantage of the new method is that by means of the method of stabilization of an individual substrate for the spectral diagnosis of malignancy, a faster and better method for determining malignancy is provided in comparison with known methods.

Examples of methods for obtaining an individual substrate for spectral diagnosis of malignancy

Example 1. Take sterile in the usual way from the human body 2,375 mg of pleural effusion or pericardial effusion or ascites as exudate and add 0.125 mg of stabilizer. As a stabilizer, a 10% aqueous solution of cesium sulfate or ammonium sulfate or gunidinium sulfate or potassium primary phosphate is used. The resulting mixture was centrifuged for 900 seconds at a speed of 25 rpm. To an aliquot of the obtained clear solution, 0.5 mg of a clear solvent of spectral purity of 95% ethanol or dioxane or tetrahydrofuran was added and mixed well. The resulting sample of the individual substrate is frozen at 253 K before use or immediately used for spectral studies of malignancy.

Example 2. Take sterile in the usual way from the human body 2,375 mg of ascites as exudate and add 0.125 mg of the stabilizer. As a stabilizer use a 10% aqueous solution of cesium sulfate. The resulting mixture was centrifuged for 950 seconds at a speed of 25 rpm. To an aliquot of the resulting clear solution was added 0.5 mg of a clear solvent of spectral purity of 95% ethanol and mixed well. The resulting sample of the individual substrate is frozen at 253 K before use or immediately used for spectral studies of malignancy.

Example 3. 2.34 mg of ascites as exudate are taken sterilely in the usual way from the human body and 0.06 mg of stabilizer is added. As a stabilizer using a 10% aqueous solution of ammonium sulfate. The resulting mixture was centrifuged for 800 seconds at a speed of 26.6 rpm. To an aliquot of the obtained clear solution, 0.6 mg of a clear solvent of spectral purity of 95% ethanol was added and mixed well. The resulting sample of an individual substrate is frozen at 250 K before use or immediately used for spectral studies of malignancy.

Example 4. Take sterile in the usual way from the human body 2.56 mg of ascites as exudate and add 0.04 mg of stabilizer. As a stabilizer, a 10% aqueous solution of potassium hydrogen phosphate is used. The resulting mixture was centrifuged for 1000 seconds at a speed of 23.3 rpm. To an aliquot of the resulting clear solution, 0.4 mg of a spectral-purity solvent of 95% ethanol was added and mixed well. The resulting sample of an individual substrate is frozen at 257 K before use or immediately used for spectral studies of malignancy.

Example 5. Take sterile in the usual way from the human body 2.58 mg of ascites as exudate and add 0.12 mg of stabilizer. As a stabilizer using a 10% aqueous solution of guanidinium sulfate. The resulting mixture was centrifuged for 1050 sec at a speed of 22.8 rpm. 0.3 mg of a spectral solvent of 95% ethanol was added to an aliquot of the resulting clear solution and mixed well. The resulting sample of an individual substrate is frozen at 248 K before use or immediately used for spectral studies of malignancy.

Example 6. Take sterile in the usual manner from the human body, 2.065 mg of ascites as exudate and add 0.135 mg of stabilizer. As a stabilizer using a 10% aqueous solution of ammonium sulfate. The resulting mixture was centrifuged for 850 seconds at a speed of 25.8 rpm. To an aliquot taken from the obtained clear solution, 0.7 mg of a spectral solvent of 95% ethanol was added and mixed well. The resulting sample of an individual substrate is frozen at 250 K before use or immediately used for spectral studies of malignancy.

Example 7. 2.36 mg of pleural effusion as exudate are taken sterilely in the usual way from the human body and 0.14 mg of stabilizer is added. As a stabilizer using a 10% aqueous solution of ammonium sulfate. The resulting mixture was centrifuged for 880 seconds at a speed of 25 rpm. To an aliquot taken from the obtained clear solution, 0.9 mg of a spectral solvent of 95% ethanol was added and mixed well. The resulting sample of an individual substrate is frozen at 250 K before use or immediately used for spectral studies of malignancy.

Example 8. 2.36 mg of pericardial effusion as an exudate are taken sterilely in the usual way from the human body and 0.14 mg of stabilizer is added. As a stabilizer using a 10% aqueous solution of ammonium sulfate. The resulting mixture was centrifuged for 880 seconds at a speed of 25.0 rpm. To an aliquot taken from the obtained clear solution, 0.9 mg of a spectral solvent of 95% ethanol was added and mixed well. The resulting sample of an individual substrate is frozen at 250 K before use or immediately used for spectral studies of malignancy.

Examples of using the obtained individual substates for spectral determination of malignancy

Example A. In a patient XY01 (1955 / m), ascites was extracted from the abdominal region and 2.2 ml of ascites (without additional treatment) was transferred to a quartz cuvette for a UV-VIS spectroscope. The UV-VIS spectrum was recorded on a Hewlett Packard UV-VIS spectrophotometer, model 8452A, in the range of 200-800 nm. Distilled water was used as a reference solution. Figure 1 shows the obtained UV-VIS spectrum. In the range from 230 to 275 nm, two well-formed and defined peaks are seen. The absorption of the peaks has a value between 3 and 4. In the range of 380-500 nm, a clearly visible flat plateau with a low absorption value of about 0.5 (Fig. 1).

This is the usual type of spectrum for human body fluid, which is a consequence of malignant diseases in the body, in this case, the UV-VIS spectrum of ascites taken clearly indicates malignancy. The clinical diagnosis established by other diagnostic methods is as follows: Adenocarclnoma ventriculi inop. Carcinosis peritonei ascites.

Example B. Patient XX01 (1932 / g) was extracted with 2,375 mg of pleural effusion from the abdominal space and at the time of extraction no clinical diagnosis was made. An individual substrate was prepared according to the method of Example 1 and transferred to a quartz cuvette for UV-VIS spectroscopy. Distilled water was used as a reference solution. The spectrum was recorded on a UV-VIS spectropometer Hewlett Packard, model 8452A, at 200-800 nm. Distilled water was used as a reference solution. Figure 2 shows the obtained UV-VIS spectrum. In the range from 230 to 280 nm, a wide band with many peaks with an absorption value of 4-5 is visible. A wide streak with many peaks is typical for the presence of benign diseases in the body. In the range from 380 to 599 nm, a high plateau appears, which is also characteristic of the presence of benign disease (figure 2). The clinical diagnosis obtained later through the remaining classical diagnostic methods is pneumonia.

Example C. Patient XX02 (1967 / g) was extracted with 19 mg of pericardial effusion from the pericardium because she suffered from heart failure. An individual substrate was prepared in accordance with Example 4 and transferred to a quartz cuvette for UV-Vis spectroscopy. The spectrum was recorded on a UV-VIS spectrophotometer Hewlett Packard, model 8452A, range 200-800 nm. Distilled water was used as a reference solution. Figure 3 shows the obtained UV-VIS spectrum. In the range from 230 to 275 nm, two well-formed and defined peaks are seen. The absorption of the peaks is between 3 and 4. In the range of 380-500 nm, a plain plateau with a low absorption value of about 0.5 is clearly visible. This is the usual type of spectrum for human body fluid, which is a consequence of a malignant disease in the body (figure 3). In this case, the spectrum of pericardial effusion removed by UV-VIS clearly indicates malignancy. In patient XX02, through various diagnostic methods, it was found that she also has Non-Hodgkin lymphoma st, IVB; st. post irrad. CNS am I / II and VCS am.

The advantage of the UV-VIS diagnostic method by removing the spectrum of the human body fluid is that the spectrum indicates a malignant process in the body, regardless of which part of the body the fluid was taken from and in which organ the malignant process develops.

The best way of carrying out the invention

2.56 mg of ascites under sterile conditions are taken as exudate from the human body and 0.04 mg of stabilizer is added. A 10% aqueous solution of potassium hydrogenphosphate is used as a stabilizer. The resulting mixture was centrifuged for 1000 seconds at a speed of 23.3 rpm. To an aliquot of the resulting clear solution was added 0.4 mg of a spectral grade solvent, 95% ethanol, and mixed thoroughly. A sample of the obtained individual substrate is frozen at 257 K before use or immediately used for spectral determination of malignancy.

Claims (1)

 The method of stabilization of an individual substrate for spectral determination of malignancy, in which the exudate taken sterile from the human body is pleural effusion, pericardial effusion or ascites and ammonium sulfate, guanidinium sulfate, cesium sulfate or primary potassium acid phosphate are used as stabilizer and as a spectral solvent use 95% ethanol, tetrahydrofuran or dioxane, characterized in that 93.8-98.5 wt.% exudate stabilize 1.5-6.2 wt.% 10% aqueous solution of stabilizer and the resulting mixture is centrifuged for 800-1050 s at a speed of 22.8-26.6 rpm, then 10.0-26.8 wt% of a transparent solvent of spectral purity are added to the obtained transparent sample, and the resulting individual substrate sample is frozen at 248-257 K before use or immediately remove the spectrum.

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