RU69866U1 - BIOCHIP - Google Patents

Abstract

The device relates to diagnostics in the field of medicine, forensics and testing genetically modified objects in products. In accordance with the technical solution, the working area of the biochip contains asymmetrically located relative to the horizontal and / or vertical axis of the working area from two to four groups of clusters of positive control and at least one group of clusters of negative control, and the number of clusters included in each group of positive and negative control ranges from 1 to 7. One of the clusters included in each of the groups of positive control is located in opposite corners of the working area, and the ratio of the total the number of clusters of positive and negative controls is from 1: 1 to 10: 1. The working area of the biochip is located on the surface of a solid carrier or on the surface of a flexible carrier or on the surface of a multilayer attached to a flexible or solid carrier, or on the surface of a solid carrier provided with a multilayer, with the possibility of forming a hybridization volume. An oligonucleotide with a known nucleotide sequence or a homologous sequence selected from sequences of marker genes belonging to the selected diagnostic type is used as a probe material in a positive control cluster, and the probe material in a negative control cluster consists of an oligonucleotide, partially complementary to a positive sequence control, or oligonucleotide, partially complementary to the sequence of one of the probe oligonucleotides. 11 items, 3 ill., 1 ave.

Description

Technical area

The utility model relates to the field of diagnostics based on biochips containing a working area on which probes with oligonucleotides or proteins are immobilized, which include probes complementary to the diagnosed markers, as well as probes of positive and negative controls.

State of the art

Biochips are known that are used to diagnose medical diseases, biochips for diagnosing the presence of GMOs in foods and feeds, biochips for testing environmental pollution, biochips for scientific research and forensic science. In the practice of diagnosis, different types of biochips are used. Biochips with high probe densities (Rava R.P. et al., 2004) are expensive and are used mainly for scientific research, since they require sophisticated diagnostic equipment in their manufacture and in the process of data acquisition and analysis. Biochips are widely used in which in the working area of the biochip there are from several units to several tens of different types of DNA probes (Zeleny R. et al., 2001) or proteins. Simpler biochips are used in small laboratories, in hospitals and in test laboratories. On their surface there are no more than twenty types of probes. Such biochips are used for medical diagnosis of specific types of diseases, to determine the presence or absence of GMOs in products (Beletsky I.P. et al., 2005, Mirzabekov A.D. et al., 2006). Biochips of low density are cheap and allow quick analysis of a large number of biochips on fairly simple equipment.

One of the serious problems that arise during mass diagnostics using cheap biochips is the reliability of the analysis, reproducibility of the results, and the possibility of transportation and storage without reducing sensitivity and reproducibility.

To control the quality of diagnostics, methods are used related to the application of separate test probes in addition to the oligonucleotide probes in each biochip cluster, which allows monitoring the quality of probe immobilization before hybridization (Kincaid R.H., 2003). However, this method requires a lot of time and materials. Known methods for immobilizing individual additional clusters with test probes on the biochip substrate surface, for example, negative control clusters (Mirzabekov A.D. et al., 2006) or simultaneously applying positive and negative control clusters to the biochip (I. Beletsky et al. , 2005).

For more successful processing of the analysis data, additional markers are introduced onto the biochip surface, indicating one (Zeleny R. et al., 2001) or two boundaries of the working area (Mirzabekov A.D. et al., 2006), which is necessary for the operation of computer programs for processing diagnostic results. Thus, improving the quality control of fairly simple biochips and improving the quality control of the diagnostic process is an urgent task, since the well-known biochips are focused on testing individual diagnostic processes.

The closest analogues of the utility model are biochips designed for food quality analysis (Beletsky I.P. et al., 2005, Mirzabekov A.D. et al., 2006). A common feature of the known types of biochips is the presence on the working surface of additional positive and / or negative controls. The disadvantage of the biochip described in the description (Beletsky I.P. et al., 2005) is the use of a large number of positive controls of various types. An increase in the number of types of positive controls involved in the PCR reaction requires the development of parameters of asymmetric multi-primer PCR and leads to a decrease in PCR efficiency due to an increase in the number of reaction steps. In addition, the placement of clusters of positive control along one boundary of the working area of the biochip can lead to errors in the interpretation of the results, since other boundaries of the working zone are not marked.

Biochips used for diagnostics can be made of different materials. The most widely used transparent materials are made of glass (Beletsky I.P. et al., 2005, Mirzabekov A.D. et al., 2006) or polymers ((Shlyapnikov Yu.M. et al. 2007, Zarytova V.F. . et al., 2004). In some cases, an identifier characterizing the parameters of the biochip is applied to the biochip.

the identifier is widely used barcode (Zeleny R. et al., 2001). In the absence of test indicators of the boundaries of the biochip, it becomes possible to incorrectly evaluate the diagnostic results due to improper installation of the biochip in the scanning device or due to incorrect selection of image sizes when using computer programs.

Three options for improper installation of a biochip placed on a transparent medium are possible. Incorrect installation is possible both due to the horizontal turn of the biochip by 180 degrees while maintaining the position of the working surface, in which the front surface is directed upwards. It is possible to turn the biochip and set its front surface down. In the third embodiment, simultaneous horizontal and vertical turns with respect to the correct position are possible. In the third embodiment, the presence of indicators located along one of the boundaries of the working area does not guarantee interpretation errors, since indicators located, for example, on the left side of the image do not change their position, while the position of the remaining clusters changes.

The biochip is known (Mirzabekov A.D. et al., 2006), in which, along with negative control clusters, fluorescent markers are used to control the position of the biochip located along two sides of the working area. The disadvantage of such a biochip design is the decrease in the working surface of the biochip due to the placement of additional fluorescent markers in the corners of the working area and the absence of positive controls and, therefore, a decrease in the functions of biochip quality control. In addition, the markers are made of a different material than the oligonucleotide probe and therefore it is necessary to change the print head when printing probes on the surface of the biochip, for example, by the drip method.

The technical task is to create the design of a new biochip with greater reliability and reproducibility of the results obtained in the field of medicine, forensics or when testing the quality of food products.

Another technical challenge is to reduce the cost of producing biochips.

These technical problems are solved by developing a new biochip design in which positive controls have two functions: a) the function of controlling the process of biochip production and its use

during the diagnostic process, b) the function of indicating the boundaries of the working area to monitor the correct installation of the biochip in the scanning device.

By choosing the material of the positive and negative control clusters, the technical solution disclosed in the utility model allows monitoring the reliability of diagnostics, including, but not limited to: checking the quality of the biochips used, checking the quality of reagents for diagnostics, checking the correct choice of the PCR and hybridization modes, checking the correctness of washing and drying biochips. At the same time, errors that occur during computer processing of data are reduced by monitoring the correct installation of the biochip in the scanning device. Cost reduction is achieved by choosing the material from which the clusters of positive control are formed, which does not require reconfiguration of the equipment when printing clusters on the surface of the biochip.

List of figures

Figure 1. The placement of positive (Q +) and negative (Q-) clusters. Where: a) an option with a minimum number of positive and negative clusters, b), c), d) options for placing groups of positive clusters presented in the form of corner and linear structures.

Figure 2. Frontal view of a biochip equipped with a cover sheet.

Figure 3. Isometric view of a biochip equipped with a cover sheet.

Description

The proposed utility model eliminates the disadvantages of known designs and increases the reliability of the interpretation of the diagnostic results.

The biological microchip for diagnosing diseases, for forensic science and for determining genetically modified objects in products contains a planar element, on the surface of the working zone of which probes complementary to the nucleotide sequences of diagnostic markers and the sequences of positive and negative control are immobilized in clusters. In this case, the working zone contains from two to four groups of clusters of positive control and at least one group of clusters of negative control, and the arrangement of clusters of positive control located in two parts of the working area of the biochip relative to the horizontal and / or vertical axis of the biochip is asymmetric with respect to friend. In addition, the number of clusters included in each group of positive and negative controls is from 1 to 7. Moreover, one

of the clusters, included in the positive control group, is located in the corner of the working area, and the ratio of the total number of clusters of positive and negative controls is from 1: 1 to 10: 1. The working area of the biochip is located on the surface of a solid carrier or on the surface of a flexible carrier or on the surface of a multilayer attached to a flexible or solid carrier, or on the surface of a solid carrier provided with a multilayer with which at least one hybridization volume is formed. An oligonucleotide with a nucleotide sequence, or a homologous sequence selected from marker gene sequences belonging to the selected diagnostic type, serves as the material of the probe included in the positive control cluster. The material of the probe included in the negative control cluster is an oligonucleotide, partially complementary to the sequence of the positive control, or an oligonucleotide, partially complementary to the sequence of one of the probe oligonucleotides.

On figa shows a structural diagram that contains a minimum number of cluster groups. In this version of the biochip, two positive controls (Q +) are placed in the corners of the working area (1) of the biochip along the diagonal of a rectangular or square field and occupy an asymmetric position relative to the horizontal (2) and / or vertical axis (3) of the working area of the biochip. Negative control (Q-), placed along the border of the working area and is located next to the positive control.

To form an asymmetric image for placement of positive controls, a combination of several groups of positive controls can be used. On Fig shows a variant of the structural layout of the positive (Q +) and negative (Q-) clusters in the embodiment in which the groups of positive clusters are placed in such a way that they form an angular structure consisting of three clusters on one side of the working area, and linear structure on the other side of the working area, forming asymmetry in the position of positive clusters relative to each other. Corner structures can be made in the form of equilateral, as shown in figb or non-equilateral angular geometric elements, with the total number of clusters constituting the corner element, choose from 3 to 11. Preferably from 3 to 7.

Linear structures can be made in the form of a column or row of clusters placed along the boundaries of the working area of the biochip. At the same time, at least one of the clusters (4, 5, 6) included in the linear or corner structures is located in the selected corner of the biochip working zone in which the linear or corner structure is placed. In the embodiment shown in FIG. 1c, when placing groups of positive control, an asymmetric image is formed using two groups of clusters of linear structure located in opposite corners of the working area and the third cluster located in the third corner of the working area. The option of placing four groups of positive clusters located in the four corners of the working area is shown in Fig.1g.

The asymmetry of the image is formed due to the different number of clusters included in different groups. The above options include, but are not limited to other options for placing positive clusters asymmetrically located relative to the horizontal and / or vertical axis of the biochip. Positive clusters perform an additional function of indicators, and allow you to identify the location of the working area on the biochip during software processing of the results, after scanning the diagnostic results.

At the same time, at least one negative control is placed along the border of the working area and is located next to the positive control, which includes, but does not limit other options for placing negative controls on the working area of the biochip.

In the utility model under consideration, an oligonucleotide with a known nucleotide sequence, or a homologous sequence selected from marker gene sequences of the selected diagnostic type, was used as the probe material of the positive control cluster.

An oligonucleotide that is partially complementary to the positive control sequence or an oligonucleotide that is partially complementary to the sequence of one of the probe oligonucleotides is selected as the material of the probe forming the negative control cluster. In the sequence of the oligonucleotide immobilized on the surface of the working zone serving as a negative control, the number and position of nucleotides that do not match the DNA sequence of the positive control are selected so that this differentiating oligonucleotide is capable of

to form and maintain an imperfect duplex with single-stranded DNA of a positive control if hybridization and washing conditions are not observed, namely, when hybridization and washing are performed under milder than optimal conditions.

The biochip can be made of a solid, flexible or multilayer material, representing, for example, a combination of a solid planar carrier with one or more flexible layers constituting a multilayer.

As the material of the solid base of the biochip, polymers, metals, ceramics, glass, mica, or combinations thereof are used. To create inexpensive biochips, it is preferable to use glass and polymers. The polymers are selected from the group consisting of polymethyl methacrylate, polybutyl methacrylate, polyvinyl chloride, polycarbonate, copolymers of methyl methacrylate and / or copolymers of butyl methacrylate with other monomers, such as styrene, acrylonitrile, etc. The thickness of the carrier element should preferably be the same and within the thickness of the slides used 0.5 mm to 5 mm.

Polymer materials can be transparent, matte, white, black or color. To measure the diagnostic results in transmission mode, the biochip material must have a light transmission of at least 80-90%. When measuring a reflected, for example, colorimetric signal, the biochip material is selected with a white reflective surface. To measure signals in fluorescence mode, it is necessary that the material of the slide or the outer surface of the multilayer has a minimum intrinsic fluorescence.

Depending on the method for detecting fluorescence, the biochip material can be transparent, made of glass or transparent plastic, for example, polymethyl methacrylate, or made in the form of an opaque material, for example, black, which has minimal reflectivity, made, for example, of polyvinyl chloride.

As the material of the flexible layer using polymers and / or metals, the thickness of the flexible layer is from 0.05 to 1 mm The flexible layer included in the multilayer is made of materials included in the group consisting of: i) opaque materials, ii) translucent materials, iii) reflective materials, iv) light-absorbing materials.

The composition of the multilayer may include: a) flexible layers provided with one adhesive layer, b) flexible layers provided with two adhesive layers and deposited on the upper and lower surfaces of the flexible layer, c) flexible layers, not

equipped with an adhesive layer, d) combinations of layers included in paragraphs a), b), c). The flexible layers forming the multilayer are arranged one above the other and connected in such a way that one of the surfaces of the upper or lower multilayer contains a self-adhesive layer that is glued to the upper and / or lower surfaces of the supporting element and to the surface of the slides by applying pressure. An adhesive or adhesive layer should create a strong bond between the surface of the slides, the surface of the supporting element and multilayers. This connection should be stable and should not be broken when the temperature of the external environment changes in the range from -10 to 60 ° C. The adhesive layer must retain its adhesive parameters under the conditions of the action of the reagents used to wash the surface of the slides or during the formation of the modifying layer on the surface of the slides, or the reagents involved in the formation of the hybridization solution.

In the process of studying the properties of polymeric materials with an adhesive layer deposited on their surface, it was found that flexible polymeric materials with adhesive layers deposited on them (made in the form of self-adhesive films) do not change their physicochemical parameters under conditions of a changing temperature in the range from -10 ° С to + 60 ° С and in the environment of hybridization buffers. This allows the use of self-adhesive films for the manufacture of many variants of inexpensive multi-chips, since the stage of applying the adhesive layer is excluded.

A biochip made of a solid, flexible, or multilayer material can be additionally equipped with a modifying layer for stronger immobilization of oligonucleotides probes and oligonucleotides of positive and negative controls in the working area of the biochip.

The surface of a solid or flexible biochip base made of polymers, glass, metals and composites can be modified with an aminosilane layer (Lefkowitz S.M. et al., 2005). The surface of polymer biochips can be modified with reagent solutions that do not create additional layers (Shlyapnikov Yu.M. et al. 2007). It is known that probes can be applied to the working surface of a biochip without surface modification (Zarytova V.F. et al., 2004) or polymerized probes in drops of an agarose gel (Mirzabekov A.D. et al., 2003). It is possible to modify the entire surface of the biochip or to modify only the surface of the working zone located inside the hybridization zone.

Variants of biochips are known, on the surface of which hybridization volumes are created using lattices (Harvey M.A. et al., 2005), or by creating sealed hybridization volumes with liquid inlets and outlets (Yuen P.K., 2002). However, the implementation of these technical solutions increases the cost of biochips. In a simpler embodiment, the biochip is made in the form of a combination of a solid carrier and a flexible multilayer provided with an adhesive layer. At the same time, one or more through holes are made in the material of the flexible layer, the boundaries of which coincide either with the boundary of the common working area of the biochip or with the boundaries of individual working areas related to individual clusters or a group of clusters. A hybridization volume is formed between the working surface of the solid biochip carrier and the walls of the hole in the flexible multilayer (Lee J. et al., 2005; Yin Li-Te et al., 2005).

In contrast to the known variants of biochips made on a solid basis (Zeleny R. et al., 2001) or made on the basis of multilayer biochips with hybridization volumes (Lee J. et al., 2005), to increase the reproducibility of diagnostic results, this useful A biochip option is proposed for the model, according to which, in addition to the asymmetric placement of positive clusters on the working area of the biochip, a cover sheet is fixed on the auxiliary surface of the biochip, a label is riveted to one of its corners. The label provides the possibility of partial or complete removal of the cover sheet from the surface of the biochip at different stages of diagnosis.

Figure 2a shows a front view of a biochip (7) equipped with a cover sheet (8) to which a label (9) is glued. The cover sheet is glued to the surface of the auxiliary zone (10) of the biochip located along the perimeter of the working area (1). The use of a cover sheet provides an increase in the reproducibility of diagnostic results, since the cover sheet protects the working area from the effects of light and moisture from the transport and storage of the biochip, and also prevents the hybridization mixture from drying out during the hybridization process.

Fig.2b shows an axonometric image of one of the options, which includes, but does not limit other solutions for the design of the biochip (7) made on the basis of a solid support element (11) with a flexible multilayer (12) mounted on the front surface of the biochip. A through hole (13) is formed in the flexible multilayer, which forms the boundaries of the working area of the biochip. A cover sheet (8) provided with a label (9) is configured to

gluing and detaching its adhesive layer from the surface of the biochip. For the formation of the adhesive layer, double-sided adhesive tape is used (14).

The device operates as follows. In the initial state, the cover sheet is glued to the auxiliary surface of the biochip and protects the deposited clusters (15) on the surface of the working area from the influence of the external environment. Before hybridization, part of the cover sheet (8) using the label (9) is disconnected from the upper surface of the biochip and open access to the hybridization volume (16). Then the hybridization solution is applied to the working area (1) of the biochip and the hybridization volume is again closed by gluing the cover sheet (8). After hybridization, the cover sheet (8) is removed using the label (9) from the entire surface of the biochip. The surface of the biochip is washed and dried to scan the diagnostic results. Installation of the cover sheet eliminates uncontrolled contamination and the formation of false results in the diagnostic process.

To increase the reliability of the results, one or more identifiers are located on the auxiliary zone of the biochip (17), which contain information about the type of biochip, the date of its release, and other parameters identifying a specific biochip with the diagnostics being carried out. Identifier data is used in processing and storage programs for diagnostic results. In the embodiment, when one identifier (17) is placed on the surface of the biochip, it is preferably placed next to one of the end edges of the biochip, preferably in the place that the operator uses to hold the biochip when it is installed in the scanning device. In the case when more than one working zone is formed on the surface of the biochip, separated by gratings made, for example, of a multilayer material, the identifiers can be located not only on one end side of the biochip, as shown in Fig. 2a, but also from several sides, including the auxiliary zone, for example, on the lateral surfaces of the biochip. Identifiers can be made, for example, in the form of a bar code or in the form of a sticker of magnetic material.

Identifier (17) may be made in the form of a label with an adhesive layer. In another embodiment, the identifier may be plotted on the surface of the multilayer before the assembly process of the multilayer biochip structure. In addition to the general identifier on the surface of the flexible layer on the auxiliary surface can be graphically displayed

additional identifiers made in the form of digital (18) and / or letter (19) images.

In another embodiment of the biochip, the multilayer may contain a reflective layer, which increases the efficiency of scanning probes with fluorescent labels, increasing the reflected fluorescence signal up to four times. Biochips are known, representing a multilayer structure, which includes a solid base on which reflective and transparent layers are placed (Weisbuch C. et al., 2004). In the invention (Lefkowitz S.M., 2003), biochips are formed on a flexible multilayer located on the front surface of a biochip in which a reflective layer is formed by vacuum or plasma spraying.

Biochips are known in which a reflective layer can be deposited on the back of a transparent biochip carrier (Matsushita T. et al., 2006). In the proposed version of the biochip, a self-adhesive film with a reflective or retroreflective coating is used as a reflective or retroreflective layer. This film is glued to a solid supporting surface, and a second multilayer is glued to its surface with the help of which at least one hybridization volume is formed, by making corresponding through holes in the second multilayer, coinciding with the boundaries of the working areas of the biochip. Thus, the combination of asymmetric placement of positive controls in the corners of the biochip working area with the possibility of creating a hybridization volume using a multilayer and with the ability to protect the working area from environmental influences and protecting the hybridization volume by attaching the cover sheet, provides a synergistic effect to increase greater reliability and reproducibility diagnostic results conducted in the field of medicine, forensics or when testing the quality of food products.

Example

Material selection and synthesis of primer oligonucleotides.

The choice of material composition for primer oligonucleotides is selected taking into account the high specificity requirements for conserved regions of the studied genes, which allows selective isolation of DNA fragments and subsequent amplification together with primer oligonucleotides using PCR directly from biological samples.

Oligonucleotides are synthesized on an automatic synthesizer (for example, Pharmacia's Gene Assembler model) using the standard amidophosphite method. Each pair of oligonucleotides for PCR specific to the diagnostic marker consists of one unlabeled (not containing any labels necessary for subsequent analysis and interpretation of the results) specific oligonucleotide primer and labeled (containing one or more labels necessary for subsequent analysis and interpretation results) of a specific oligonucleotide.

Labels that modify primers at the 5'-end are selected from the group consisting of: catalytic, ligand, fluorescent and radioactive, which is introduced by chemical methods or enzymatically. As catalytic labels, hemin, cyancobalamin, or flavin are used (Beletsky I.P. et al., 2005). As ligand labels, biotin, dioxigenin or dinitrobenzene are used. Su3, Su5, Su7, FAM, TAMRA, R6G, R110, ROX or JOE are used as fluorescent labels. The tag lists provided include, but are not limited to, other tag options that can be used to detect a signal.

Oligonucleotides are synthesized on an automatic synthesizer (for example, Pharmacia's Gene Assembler model) using the standard amidophosphite method. The oligonucleotides that make up the probes contain a 5'- or 3'-terminal group, ensuring their immobilization on the surface of the chip, for example, a 5'-terminal phosphate group, which is introduced by chemical phosphorylation during synthesis.

Industrial applicability

The new technical solution allows the diagnostic monitoring process to be implemented including, but not limited to: checking the quality of used biochips, checking the quality of reagents for diagnostics, checking the correct choice of PCR and hybridization reaction modes, checking the correctness of washing and drying of biochips, while reducing errors that occur during computer processing of data due to improper installation of the biochip in the scanning device. The reduction in the nomenclature of materials from which the positive control clusters are made, and their use as indicators of the boundaries of the biochip working area, reduces the costs of manufacturing biochips and the costs associated with the analysis of diagnostic data.

Literature

1. Rava R.P. et al. Methods for concurrently processing multiple biological chip assays. US Patent 6,720,149 (April 13, 2004).

2. Zeleny R. et al. Automatic imaging and analysis of microarray biochips. US Patent 6,215,894 (April 10, 2001).

3. Beletsky I.P. etc. A set of primers for the detection and / or identification of transgenic DNA sequences in plant material and its containing products (options), a primer (options), a pair of primers (options), a method for detection and / or identification using them (options) and a device to implement the method. Patent RU 2265668 (2005.12.10).

4. Mirzabekov A.D. et al. A method for identifying transgenic DNA sequences in plant material and products based on it, a set of oligonucleotides and a biochip for implementing this method. Patent RU 2270254 (2006.02.20).

5. Kincaid R.H. Apparatus and methods of detecting features on a microarray. US Patent Applic. 20030186310 (October 2, 2003).

6. Shlyapnikov Yu.M. et al. Method for producing DNA chips (variants) Patent application RU 2005125776 (2007.02.20).

7. Zarytova V.F. et al. A method for producing DNA chips. Patent RU 2236467 (2004.09.20).

8. Lefkowitz S.M. et al. Chemical arrays. US Patent 6,841,663 (January 11, 2005).

9. Harvey M.A. et al. Device for preparing multiple assay samples using multiple array surfaces. US Patent Applic. 20050135974 (June 23, 2005).

10. Yuen P.K. Well frame including connectors for biological fluids. US Patent Applic. 20020168624 (November 14, 2002).

11. Lee J. et al. Patch for microarray reaction chamber having adhesive means support and two or more adhesive materials. US Patent Applic. 20050136459 (June 23, 2005).

12. Yin Li-Te et al. Biochip containing reaction wells and method for producing same and use thereof. US Patent Applic. 20050106607 (May 19, 2005).

13. Weisbuch C. et al. Biochip type device. US Patent Applic. 20040222480 (November 11, 2004).

14. Matsushita, T. et al. Component, apparatus, and method for analyzing molecules. US Patent 6,999,166 (February 14, 2006).

15. Mirzabekov A.D. et al. Method for the polymerization immobilization of biological macromolecules and a composition for its implementation. Patent RU 2216547 (2003.11.20).

Claims (11)

1. A biological microchip for the diagnosis of diseases, forensic science and testing genetically modified objects in products, containing a solid carrier, on the working area of which in the form of clusters oligonucleotide probes are immobilized, complementary to the nucleotide sequences of diagnostic markers and sequences of positive and negative control, characterized in that the working area contains from two to four asymmetrically located relative to the horizontal and / or vertical axis of the working area ex groups of clusters of positive control and at least one group of clusters of negative control, and the number of clusters included in each group of positive and negative controls is from 1 to 7, while one of the clusters included in each of the groups of positive control, placed in opposite corners of the working area, and the ratio of the total number of clusters of positive and negative controls is from 1: 1 to 10: 1, where the working area of the biochip is located on the surface of a solid carrier or on the surface of a flexible carrier or on the surface of a multilayer mounted on a flexible or solid carrier, or on the surface of a solid carrier provided with a multilayer, with the possibility of forming a hybridization volume, and an oligonucleotide with a known nucleotide sequence or homologous is used as the probe material included in the positive control cluster a sequence selected from sequences of marker genes related to the selected type of diagnosis, and the probe material included in the cluster a negative control consists of an oligonucleotide, partially complementary to the sequence of the positive control, or an oligonucleotide, partially complementary to the sequence of one of the probe oligonucleotides. 2. The biochip according to claim 1, characterized in that on the front surface of the supporting element an additional covering sheet is installed, provided with an adhesive layer and a label around the perimeter, with the possibility of attaching or detaching part or all of the covering sheet from the auxiliary zone of the supporting element. 3. The biochip according to claim 1, characterized in that the groups of positive clusters are made in the form of a corner or linear structure. 4. The biochip according to claim 3, characterized in that the corner structures are made in the form of equilateral or non-equilateral corner geometric elements. 5. The biochip according to claim 1, characterized in that polymers, metals, ceramics, glass, mica, or combinations thereof are used as a solid base of the biochip. 6. The biochip according to claim 5, characterized in that the polymers are selected from the group consisting of polymethyl methacrylate, polybutyl methacrylate, polyvinyl chloride, polycarbonate, copolymers of methyl methacrylate and / or copolymers of butyl methacrylate with other monomers such as styrene, acrylonitrile. 7. The biochip according to claim 5, characterized in that the thickness of the solid base lies in the range from 0.5 mm to 5 mm. 8. The biochip according to claim 1, characterized in that polymers and / or metals are used as a flexible layer. 9. The biochip according to claim 1, characterized in that the flexible layer is made of polymers and / or metals, the thickness of the flexible layer being from 0.05 to 1 mm. 10. The biochip according to claim 5, characterized in that the solid base is made of opaque or translucent materials. 11. The biochip according to claim 9, characterized in that the flexible layer is made of materials included in the group consisting of: i) opaque materials, ii) translucent materials, iii) reflective materials, iv) light-absorbing materials.