METHOD FOR DETECTING MALIGN PROLIFERATION

Abstract: A method for detecting malign proliferation tumorous diseases including leukemia. The method is accomplished by optically examining blood serum, body fluid, cell agglomerates or tissue preparates having previously been dried - preferably under physiological conditions - and the pathological change is proved if at least partial lack of the terminal effect is observed.

Description: METHOD FOR DETECTING MALIGN PROLIFERATION Technical field The present invention relates to a diagnostic method for detecting malign proliferation - tumorous diseases , including leukemia - from blood serum, body fluid, cell agglomerates and tissues . Background art It is -well known that the essential of successful treatment of malign tumorous diseases is the diagnosis thereof in a very early state . Extended research has been carried out recently for develop¬ ing such diagnostic methods . These methods are generally based on the measurement of the concentration or the change in concentrat¬ ion of certain specific proteins - e . g. enzyme - in the blood. These methods are , however, of controlled area of use , and need longer time and cost. No method has been disclosed so far which would generally be applicable in the diagnosis of tumorous diseases. Therefore , the present invention aims to provide a simple and quick method for detecting tumorous diseases , possibly in a very early state thereof. D sclosure of invention The present invention is based on the recognition that in the samples of blood serum, body fluids , cell agglomerates and tissue preparates prepared suitably under physiological conditions , charac¬ teristic protein filaments can be observed by optical methods. These filaments are characterized by the so-called "terminal effect", i^ e . in microscopic picture the ends of the said filaments in every case reach or advance in the possible shortest way towards a point of another - 2 - filament provided that the sample originates from a healthy organ. If the sample has been obtained from tumorous organ, the "terminal effect" is not characteristic, i. e . the filament system characteris¬ tic to the healthy state does not occur at all or the filaments meet each other partly only. The characteristic features of the terminal effect mentio ed above and the formation thereof is explained herebelow in the example of blood serum. It is obvious , however, that by making pre¬ paration from samples of other body fluids, cell agglomerates or tissues, the appearance of terminal effect will be alike . Brief Description of drawings In the following s the invention is demonstrated by the .figures more in details. Fig. 1 is a photograph of dried blood serum taken by electron ∞-croscope . Fig. 2 is a photograph of blood serum prepa ate of healthy patient taken by electron microscope . Fig. 3 is the photograph of blood serum preparate of tumorous patient taken by electron microscope. Eig. 4 is the photograph of the filament structure of blood serum preparate from a healthy patient. Fig. 5 is the photograph of the filament structrure of blood serum preparate from a tumorous patient. Fig. 6 is a section of the photograph of blood serum preparate from healthy patient showing the terminal effect. Fig. 7 is the photograph of Ascytes fluid preparate from a healthy patient. Fig. 8 is the photograph of Ascytes fluid preparate obtai from a tumorous organ. When blood taken venally is allowed to sediment and the the serum is dropped on microscope slide , a characteristic picture is obtained after drying under mild conditions . Figure 1 shows interesting formations , filaments in the crackled preparate . It has been found that the picture of blood serums from healthy and tumorous organism, respectively, are basically different after drying. Fig. 2 shows the dried blood serum preparate of a healthy patient while Fig. 3 shows the same from a patient suffering from malign tumour . The splits shown on the photographs always form along the filaments ; when the upper layer of the preparate is removed, the filament structures according to Figs . 4- and 5 are obtained. Fig. L shows clearly that the filaments are in close contact with each other, i. e . the terminal effect. shown in Fig. β more in details^ occured. Fig. 5 shows no terminal effect, neither filaments were formed; the structure refers unambiguously to the presence of malign processes. Similar pictures may be obtained when examining the preparates of Ascytes fluids instead of blood serum preparates . Figs. 7 and 8 show the picture of these preparates obtained from healthy and tumorous organism, respectively. It can clearly be seen from Fig. 7 that in case of healthy cells intercellular contact was formed by the filaments and the charac¬ teristic splits occured along these . Fig. 8 shows the lack of terminal effect, i. e . the presence of malign processes in the organism. The method according to the invention was worked out on basis of our aforesaid experiences . The method comprises optically examining blood serum,body fluid, cell agglomerate or tissue preparates having been dried preferably under physiological conditions and the pathological change is proved on basis of at least partial lack of the terminal effect. The preparates can be obtained generally by methods known per se . It is preferred, however, to prepare them under controlled - - conditions as the method can be standardised by this way and the different samples can be compared easier. In case of examining blood samples , the preferred process is as follows: Native blood is taken, the blood serum is prepared suitably by sedimenting or centrifuging, then the serum is dropped on a suitable support. Good preparate can be obtained when applying a drop of the volume of 1 to 20 Λil or about the same amount is distributed on microscope slide . Drying of the fluid is generally carried out at controlled temperature , preferably at 20 to 50 C by maintaining constant moisture content and pressure . The drying can be accomplished by using vacuum, however, it is preferred to work at atmospheric pressure . Drying may also be performed by freezing to the critical point when the water in the sample separates in micro crystalline form and the splits will again follo the filaments. Examination and evaluation of the preparates are made on basis of the terminal effect. The organism can be regarded as healthy if the terminal effect can be found in the whole of the sample. If the terminal effect is lacking - even if only in part - , the phenomenon refer to malign process. Evaluation is accomplished visually as the most simple way, however, it is also possible to use special apparatus and methods , respectively, which might ease the identification. Processes like these include Urbane sek-algorhythm method, Fourier-transformation algorhyth the method of closed graphs , counting the black /white /black or reversed transitions or any combination thereof. However, it is to be understood that the mode of evaluation is not- limited to the above anyway and the choice of the evaluation method does not form a part of the scope of protection. - 0 The diagnostic method according to the invention was controlled in tests carried out on more than 1000 patients and was proved to be effective with a very good accuracy in determining tumorous states of different type . The method is very advantageous as it is very quick, thus it can be used for screenings , for following therapies , as well as in the search for anti-tumour compounds for quick in vitro screening of the effect of test materials . Example A group consisting of 116- + patients was tested with the method according to the invention. During the tests the results obtained from the pictures of dried blood serum were compared with clinical hysto logical diagnoses . When evaluating, three sub-groups were formed as shown in Table II. The first group consisted of patients whi ch were proved to have malign tumour according to hystological examination. The method according to the invention /MTA/ showed malignity in 358 of the 383 case /94- %/ . Table I herebelow shows the distribution of different malign diseases within this group. T a b l e Malign disease Number of cases Leukemia 74 Malignant lymphomes 93 Head and neck cancer 9 Lung cancer ' 17 Malignant breast neoplasms 4-1 Malignant neoplasm of digestive 48 organs Malignant neoplasm of female genital 12 organs Malignant neoplasm, of male genital 56 organs Brain tumour 7 Neoplasm of urinary system 20 Epithelioma 6 The second group served as control, consisting of healthy patients below an age of 25. The method according to the invention found 230 from the 242 of the group to be healthy /95 %/ . The third group consisted of patients of an age between 25 and 85 who did not suffer from tumorous disease according to hystological examination but had other illness , such as hepatitis , pneumonia, hypertonia, diabetes , benign tumour , etc . The MTA prove 376 cases as negative from the total of 539 /70 %/ . As mentioned, t this group clinical diagnosis has not detected malign tumorous disease but the possibility could not be eliminated. A summary of the above test is shown in Table II here- below. T a l e II Group Total of MTA MTA MTA patients + total % Hystologically proved 383 358 25 94 tumorous patients Healthy control groups 242 12 230 95 /under the age of 25/ Mixed group /no clinically detected cancer but possi¬ bility , cannot be eliminat¬ ed/ 539 163 376 70 It is believed on basis of the results obtained at the first two groups that MTA detects the malign processes in their very early state , especially when there are no serious symptoms and malignant tumour cannot be detected or only in certain cases by other clinical methods . From the patients of the "mixed group" proved to be positive by the method according to the present invention 7 were later proved to have malign tumour by subsequent clinical examinations while the others were kept under ad ditional ob servation .

Claims: Claims 1. Method for detecting malign proliferation - tumorous diseases including leukemia -, characterized by optically examining blood serum, body fluid, cell agglomerates or tissue preparates having been dried preferably under physiological condit¬ ions and the pathological change is proved on basis of at least partial lack of the terminal effect. 2. The method according to claim 1, characterized by using dried blood serum. 3. The method according to claim 1, characterized by using dried Ascytes fluid.

Inventors: KOVACS ADAM, KOVACS, ADAM
Applicants: KOVACS ADAM, KOVACS, ADAM
Application year: 1985
Main classification: G01N33
IPC codes: G01N33