"System and method for solid phase analysis of biological samples"

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The present invention is relative to a system and a method for solid phase analysis of biological samples.

In particular, this invention is relative to a system and method for solid phase analysis of biological samples, of the type used to perform immunoassays on a solid phase, for analytes present in a liquid solution, especially for in vitro diagnosis of allergies and autoimmune diseases.

As it is known, many types of reaction systems for performing sample analyzes are currently known. In some known systems, a reaction system has a plurality of reaction chambers grouped in the same device and arranged each other with a predetermined order. Systems having reaction chambers are used in combination with fully automatic equipment in which a large number of samples are analyzed, and a possible application of these devices is in vitro analysis of allergies.

Systems for solid phase analysis of biological samples can be used in combination with fully automatic equipment in which a large number of samples are analyzed, said equipment being configured to: dispense the samples under examination in the reaction chambers;

- wait the time necessary for the chemical reaction to take place;

remove by appropriate washing, by mean of an introduction from a first end of a U-chamber duct and suction from the other end of the duct of the unreacted solution;

- feed a tracer reagent;

- wait for the time necessary for the second chemical reaction;

repeat the washing procedure of the unreacted solution;

- feed an appropriate detection solution;

- wait for the development of the reaction and read the calorimetric or energy variation produced.

In use, the analytes present in biological fluids are generally determined by specific immunological reactions between antibody and antigen that form an immuno-complex. The reagent that reacts specifically with the analyte is marked so as to allow the detection and measurement of the analyte concentration itself. The marking can be made with radioactive, fluorescent, chemo-luminescent substances or by enzymes. In the latter case, the signal relative to the enzyme must be made explicit by adding a suitable substrate which must modify its state so as to become detectable (for example by changing color and energy state) . To facilitate the separation between the reacted analyte and the unreacted analyte (so it is not specifically related to the test performed) different solid phases are used. The binding of the specific reagent to the selected solid phase, allows to easily separate the immune complex that is formed between the specific reagent and the analyte. It is possible to use a large number of solid phases, such as: paper discs, plastic sphere, microplate wells, test tubes, cellulose capsules, etc.

A known system is described in patent application US2012/0283122A1, in particular it describes an optimized method, Multiple Microarrays, for dosing multiple and distinct analytes by standardizing an internal calibration curve and a calibration curve with known standards. Two calibration curves are generated during the process of various analytes determination. The system and the method described, uses fluorescent antibodies, each having different wavelength fluorescence, measured by scanner.

Another known method is described in patent application US2011/0306511. The method described concerns dosage and quantification of analytes, distinguishing each analyte, in a single reaction vessel. Two calibration curves are generated during the process of various analytes determination. The method uses different antibodies conjugated with different fluorescence that have different wavelengths for a subsequent identification. Therefore, containing the identification of different analytes in the same reaction chamber.

A system for performing diagnostic assays for allergies and autoimmune diseases is described in WO2014/145581A4. In this patent application, a method and a system are disclosed to improve and overcome some critical issues, in which the analytes present in the biological samples bind to paramagnetic particles for their identification .

Also, methods are known in which it is possible to perform a recalibration of reference curve and curves for families of allergens and / or single allergens on the basis of calibration parameters which can derive from different sources.

Although beneficial in many ways, these systems suffer from the fact that in known reaction chambers the liquid is insert and then aspirated, this procedure, due to the reaction chamber conformation, causes a residual liquid in the reaction chamber, which reduce sensitivity and specificity of the tests.

Purpose of the present invention is to provide a system and method for solid phase analysis of biological samples allowing to optimize a reaction vessel so as to make the production and application phases of the solid phase internal to each reaction chamber, and therefore having characteristics such as to overcome the limits which still influence the systems previously described with reference to the prior art.

Also, purpose of this invention is to provide a method for solid phase analysis of biological samples which guarantees control and security of a quantitative analysis result, by checking and verifying a reference curve.

According to the present invention, a system for solid phase analysis of biological samples is provided, as defined in claim 1.

According to the present invention, a method for solid phase analysis of biological samples is provided, as defined in claim 11.

For a better understanding of the present invention, a preferred embodiment is now described, purely by mean of non-limiting example, with reference to the attached drawings, in which: - figure 1 shows a section view of a system for solid phase analysis of biological samples, according to the invention;

- figure 2 shows another section view of the system for solid phase analysis of biological samples, according to the invention;

- figure 3 shows a top view of the system for solid phase analysis of biological samples, according to the invention;

figure 4 shows a section view of the system for solid phase analysis of biological samples in figure 4, according to the invention;

- figure 5 shows another section view of the system for solid phase analysis of biological samples of figure 4, according to the invention;

figure 6 shows a schematic view of a system according to the invention;

- figure 7 shows a flowchart of a method for solid phase analysis of biological samples, according to the invention;

- figure 8 shows a flowchart of a method for solid phase analysis of biological samples, according to the invention . With reference to this figure, and in particular to Figure 1, a system for solid phase analysis of biological samples is shown, according to the invention.

More in detail, the system for solid phase analysis of biological samples comprises:

- At least a reaction chamber 100 consisting in a vertical U-shaped capillary duct;

- First feeding means suitable to feed a liquid to be analyzed, i.e. an analyte under examination, to the reaction chamber 100 for reacting said liquid with the substances contained in the solid coating, in order to generate immune complexes;

- a washing unit of said reaction chamber 100, able to eliminate the analyte under examination and / or additional reagents fed to the reaction chamber at later times in the analysis process;

- second feeding means suitable to feed an additional immuno-enzymatic reagent, for example an enzyme conjugated to the specific antibody for the test analyte ;

- third feeding means suitable to feed a detection reagent which determines an optically and detectable modification of the reactants in the reaction chamber

100, in particular of the immuno-enzymatic reagent; - at least one photometric sensor that reads the intensities of the optical modifications of the immuno- enzymatic reagent during the reaction with the detector reagent ;

- at least one processing unit configured to determine by interpolation with a reference curve, the quantitative numerical values of the allergens and / or antibodies present in the reaction chamber from the photometric data;

- a control unit configured to execute a software program in which are coded the instructions for controlling said feeding means and said units corresponding to the steps of the method according to the invention.

According to an aspect of the invention, the reaction chamber 100 comprises an element having at least one of its walls covered with a solid material in which one or more different substances are contained, as single allergens or autoantigens, or groups or families of allergens or autoantigens .

According to another aspect of the invention, the reaction chamber 100 comprises boundary walls having al least a portion of inner surfaces coated with a solid phase layer containing one or more substances, as single allergens or autoantigens, or groups or families of allergens or autoantigens.

According to an aspect of the invention, at least one of the reaction chambers of the system, preferably each reaction chamber provided in the system, is formed by two parallel and spaced apart ducts, each duct being a branch of the U-shaped reaction chamber. Each duct is open at both ends .

The system further comprises a removable cap 105 for closing the ducts, which can be sealed to one of the two ends of the two parallel tubular ducts. According to an aspect of the invention, the cap 105 is provided with a specific color univocally related to a specific substances group which can be linked to the solid phase applied or expected in one of the branches of the U-shaped duct, such as different families of allergens or different groups of autoantigens .

According to a preferred embodiment, two rectilinear ducts forming the branches of the U-shaped reaction chamber 100 end with their lower end in a common chamber intended to form a sealing fixing seat of a common closure cap of said chamber. The reaction chamber 100 is like a cup while the closure cap 105 has a surface facing the ends of the two ducts that form the branches of the U- shaped reaction chamber 100.

The cap 105 and the chamber take a relative position of end-of-stroke for mutual coupling. In this end-of-stroke position the chamber is sealed together and forms, together with the inner surface of the cap, a transverse conduit connecting the two ducts, forming the two branches of the U-shaped chamber.

According to an aspect of the invention, a reaction container device has a plurality of U-shaped reaction chambers which are arranged aligned with respect to each other, along the plane defined by U-shaped branches axes. The chambers are spaced from each other in a predetermined extent .

According to an aspect of the invention the liquid flow in the U-shaped reaction chamber is unidirectional, passing through the first and the second branches. This conformation of the reaction chamber, advantageously, allows to optimize the sensitivity and specificity of the process .

Optionally, the lower closing cap of said chambers is constituted by a plurality of side-by-side caps each destined for a U-shaped reaction chamber 100 in which the plurality of caps 105 is integrated in a single block. Advantageously according to the invention, the colors provided for the caps 105 allow to differentiate the reaction containers of the system on the basis of the substances contained in the solid phase and, therefore, to the analytes that the reaction in each chamber should detect. Therefore, the execution of an analysis aimed at performing immunoassays and identifying and diagnosing allergies or autoimmunities, is facilitated since the service personnel chooses the reaction container having a staining cap corresponding to the family of allergens or groups of autoantigens examined, avoiding errors.

Advantageously, the invention makes it easier to read the test results with reference to the identification of the substances to which a patient's serum has been positive. Since the reading of the test results is done by photometry, the equipment intended to carry out the steps of the procedure are provided with sensors that allow the reading of the colors. It is possible to automate the editing of test results by automatically recognizing the color of the caps associated with the reaction devices.

According to an aspect of the invention, the caps are made of opaque material while the channels forming the U- shaped chambers are made of transparent material, in order to allow photometric readings. According to an aspect of the invention, the groups of allergens are subdivided and associated with a color according to the following criteria: gramineous - blue; grasses - light green; arboreal plants - dark green; mites - dark orange; house dust - light orange; epithelial - Red; molds and yeasts - light blue; pests, insects, professional poisons - gray; food - Yellow; mixtures of allergens - pink; screening test - violet; allergenic components - purple .

Advantageously according to the invention, it is easy to follow the production steps of the reaction container devices with reference to the application in the containers of the solid phases. In fact, having available branches of straight U-shaped channels open at their opposite ends.

Techniques for applying solid phases to the walls of a reaction chamber are known. Generally, walls of the U- shaped chamber are made of a plastic material selected from one or more of the following: plastic polymers such as polystyrene, polyvinyl chloride, polybutadiene, etc., polystyrene-polybutadiene copolymer or glass or a combination of said materials. The walls of at least one branch are coated with the reactant for example by the use of proteins. The coating steps can be preceded by activation steps by, for example, treatment with gamma rays or by plasma treatment or other treatments aimed at improving the adhesion of coating layers.

It is clear that the manipulations necessary for the coating of a duct open on both ends is much easier than that of a practically blind duct, except for a transverse cross-section .

Advantageously according to the invention, the two- part construction of the U-shaped chambers also has advantages in terms of manufacture. The two branches can be easily made by drilling further, the realization of ducts open at the two ends avoids the formation of undercuts which would make a manufacturing process by injection molding techniques complex.

According to an aspect of the invention, the system comprises a plurality of reaction chambers 100 in particular lined and adjacent between them, each having a U shape .

Each U-shaped reaction chamber comprises a first and a second vertical branches 101, 102 and a transverse duct 103 which connects the lower ends of the two vertical branches 101 and 102 together, putting them in fluid communication.

The plurality of reaction chambers 100 are aligned along a vertical plane, sharpened and equally spaced, and with the axes of the vertical branches 101 and 102 of the reaction chambers 100 contained in the same median vertical plane which, with reference to the example illustrated, coincides with the plane V-V of figure 4.

The first and the second branches 101, 102 which form the reaction chambers 100 are preferably rectilinear and parallel, ending in a recess 104 with an elongated cross- section, slot-shaped.

The recess 104 is open on the bottom side of the system and constitutes a sealing seat for a lower closure cap. Said lower closure cap forms, together with the bottom wall of the recess 104, a transverse duct 103 which connects the ends of the two branches 101 and 102 which open into the lower recess 104.

As shown in the figures, each reaction chamber 100 has the same shape as other reaction chambers, and therefore the recesses 104 are identical and open on the bottom side with an identical opening, while the openings are spaced apart .

The caps 105 are provided in a number corresponding to that of the reaction chambers 100 and are part of a single closure body 115 all made in the same material and shaped so as to form individual caps having portions intended to be inserted in the terminal recesses 104 of each reaction chamber 100, while the peripheral walls delimiting such recesses interpenetrate cuneiform cavities intermediate between the individual parts of the caps 105.

According to an aspect of the invention, at the two opposite ends of each row of reaction chambers 100 there are provided means for detachable mutual coupling of an adjacent system which can be provided hooked to one or the other end of a system.

According to another aspect of the invention, the system comprises at a first end a vertical pin 110, and at an opposite end 120 it comprises a tubular seat. The pin 110 have a cross-section and a diameter such as to engage the tubular seat.

According to a preferred embodiment, both the pin and the tubular seat are provided with a truncated cone-shaped surface and are intended to be inserted into one another when two systems are coupled together and aligned according to the longitudinal axes along which they are aligned the reaction chambers 100.

According to an aspect of the invention, the caps 105 are tapered with a truncated cone shape like the recesses 104, and the recesses have at their ends a seat 124 for a further engagement destined to constrain an eccentric axial protrusion 125 of the corresponding caps 105. Advantageously the protrusion 125 and the seat 124 allow a greater coupling force generated between the cap and the recess .

The solid phase of the reactants is at the inner surface of the cavities formed by the ducts forming the reaction chambers 100. Preferably, the reaction chambers are made of Styrolux which is an optically transparent plastic, and is a polystyrene-polybutadiene copolymer. Said material can be easily coated with the solid phase, for example by using proteins, similarly to what happens for wells of a polystyrene microtiter plate.

The reaction chambers 100 can be made alternatively, in materials such as plastic polymers (polystyrene, polyvinyl chloride, polybutadiene, etc.) or glass. For particular purposes, the internal surfaces of the bodies can be activated by treatment with gamma rays, or by plasma treatment or other suitable treatments.

According to a preferred embodiment, the system consists of a parallelepiped device containing five vertical U-shaped reaction chambers 100. The reaction chambers 100 have relatively small diameters and an internal volume of less than 50 mΐ . Said internal volume is lower than the volume of a common microtiter plate wells. Preferably, the parallelepiped device containing U-shaped reaction chambers 100 has height equal to 15 mm, comprising the cap in its lower part, and it has height equal to 24 mm. the thickness of the parallelepiped device is equal to 3mm in its upper portion and equal to 4mm in its lower portion. The distance between a reaction chamber 100 and a next adjacent reaction chamber 100 is preferably 4.71 mm.

The parallelepiped device has a cylindrical vertical duct at each end, in order to indicate the positioning and inserting direction of the system in the in the analytical platform of an automatic dosing instrument.

According to an aspect of the invention, the first vertical branches 101 has a diameter smaller than the second vertical branches 102. The transverse duct 103 allows the passage of liquids from the first branches 101, being a duct with a smaller diameter, to the second branches 102, being a duct with a larger diameter 102.

According to an aspect of the invention the system comprises a plurality of reaction chambers 100, made of a transparent rigid material, and a cap 105, made of an opaque elastic material, that can be coupled inserting the protrusion 125 in the seat 124. The coupling operation is facilitated by the fact that the cap 105 is made of polyethylene added with a softener, resulting flexible and compressible . The system for solid phase analysis of biological samples according to the invention is intended for use in an automatic analysis system. The system allows various reagents to be introduced for the assay, including the washing solution between the different steps of the assay, through the first vertical branches 101 of smaller diameter. Instead, the second vertical branches 102, having a larger diameter, is suitable for the suction of fluids after the relative incubation times and also for the photometric reading of the final results of the dosage.

In use, once the system has been placed in the in the test instrument platform, on the coupling pin 110 the tubular element 120 of the subsequent device is inserted. A tubular seat 120 remains free at one end of a row of mutually coupled devices, and a truncated-cone pin 110 remains free at the other end. The tubular seat 120 is supported by a rigid metal driven element which is fixed to the edge of the platform.

Even if the system according to the invention has wide applicability in the field of the detection of analytes in a fluid, it is particularly suitable for immunoassays in solid phase ELISA and analogous immunoassays in solid phase invention. Once the reaction chambers 100 are coated by appropriate procedures with the desired proteins (or other) , the proteins (or other) bound to the inner walls remain stable for a few years. The groups of allergens or autoantigens to the walls of the reaction chambers 100 are advantageously indicated by the color of the caps which is uniquely associated with a specific group of these substances .

As stated above, this invention also refers to a method for solid phase analysis of biological samples, with immunochemical methodology, especially in order to allow in vitro diagnosis of allergies and autoimmune diseases.

The method for solid phase analysis of biological samples according to the invention, includes the steps of: a) Providing at least a reaction chamber, comprising an element having at least one wall covered with a layer of solid material in which one or more different substances belonging to individual allergens or autoantigens or to groups or families thereof are contained, alternatively the reaction chamber has at least a portion of an inner wall having a solid phase coating and containing one or more substances belonging to single allergens or autoantigens or to groups or families of the same;

b) feeding to said reaction chambers a liquid to be analyzed and reacting said liquid with substances contained in the solid phase coating in order to generate immuno-complexes ;

c) eliminating the residual analyte at the end of a predetermined reaction time;

d) washing the reaction chamber and feeding into the reaction chamber a further immuno-enzymatic reagent, for example an enzyme conjugated to the antibody specific for the analyte, able to react with the solid phase present in the reaction chamber;

e) eliminating from the reaction chamber unreacted part of said further reactant, at the end of a predetermined reaction duration.

f) measuring the amount of said immune-enzymatic reactant that reacted with the substances present in the solid phase, in which this measurement takes place by means of a detection reagent which determines an optically detectable modification, this modification being measured by photometry;

g) determining the concentration of said immuno-enzymatic reagent specific for the allergen present in the analyte, by interpolating intensity values measured photometrically with a predetermined reference curve. According to an aspect of the invention, the method, together with the execution of the analysis procedure and the succession of analysis steps, also includes the following steps:

h) providing at least one second reaction chamber identical to the first reaction chamber prepared for the analysis of the analyte under examination and which is subjected together with said at least one reaction chamber, in succession or in parallel to steps b) and e) ; and in which

i) feeding to the step b) a control analyte having a known composition, in particular in relation to the antibodies and to the allergens present therein;

j ) comparing values obtained from step g) with known values for the control analyte and recalibrating or modifying the reference curve obtained, based on the difference between the measured values and the known values;

k) applying automatically a new reference curve for a repetition of step g) in relation to the results of the analysis of the analyte under examination referred to in step b) .

According to an aspect of the invention, step j ) , of feeding a control analyte, is preceded by the comparison of the values obtained from the analysis of the control analyte with values obtained from the analysis of the control analyte with maximum and minimum values of the admissible difference between measured values and known values. Step j) is executed when this difference is within said values and an alarm is generated when the difference exceeds the maximum dream value and is lower than the minimum one .

The method according to the invention, assumes to have a single reference curve that links the intensity of photometric measurements to the concentration of an immuno- enzymatic reagent linked to a specific allergen. Since the characteristics of the reagents fixed in the reaction chamber and intended to react with the liquid solution to be analyzed, the respective further reactants, and the specific characteristics of photonic transmissibility of the reaction chamber are variables, the reference curve is a critical element of the result determination process. Currently, the reference curve is indicated by the supplier of the reaction chambers previously provided with the solid phase reagents. The reference curve can therefore be set in the photometric data processing process which is performed by software performed by a processing unit.

According to an aspect of the invention, a further verification analyte is included for which a dedicated reaction chamber is provided which is identical to that provided for the analyte under examination and for the control analyte and which is subjected to the same steps as the control analyte in succession or in parallel at the same. The quantitative results obtained from the analysis are compared with the known values of the verification analyte. The difference between the values obtained from the analysis and the known values is compared with the minimum and maximum threshold values for the difference and the difference is further compared with the difference of the measured and known values calculated for the control analyte. In this case, step j) can be inhibited and at least one alarm can be generated if the difference between the values measured with the analysis and those known for the verification analyte exceeds said maximum threshold or is lower than said minimum threshold. This consequence can also occur if the difference between the measured value and the known value for the control analyte is either less than a maximum or minimum threshold with respect to the difference between the measured value and the known value for the verification analyte. The method is preferably applicable in combination with the system according to the invention or by providing five reaction chambers or multiples thereof for the test analyte and at least two to five reaction chambers for the control analyte and at least one reaction chamber for the verification analyte. According to a preferred embodiment of the method, the reaction chambers are in groups of five arranged in a row between each other, each group being connectable to one or more further groups .

Said method is suitable to be implemented by mean of the system described and comprising: at least one reaction chamber in which an element is present whose wall is covered with a layer of solid material in which one or more different substances are contained belonging. single allergens or autoantigens or to groups or families of the same Or presenting the said chambers at least one part of the inner surface of at least one of its boundary wall a covering layer of the said solid phase surface and containing one or more different substances belonging individual allergens or autoantigens or groups or families of the same; feeder organs a liquid from, to analyze, or an analyte under examination, to said chamber reaction to react the said liquid with the substances contained in the investment in the solid phase in order to generate immune complexes; washing unit of said reaction chamber for the elimination of said analyte under examination of further reactants fed to said reaction chamber in successive times of the analysis process; power supply organs of a further agent immuno-enzymatic as an enzyme conjugated to the antibody specific for the analyte in examination: feeding means a detection reagent which determines an optically detectable modification of the reactants in the reaction chamber, in particular of the immuno-enzymatic reagent; at least a "photometric" sensor that reads the intensities of the changes: optics of the immuno-enzymatic reagent when reacting with the detector reagent; at least one processing unit which is configured to determine by interpolation with reference curve the quantitative values of allergens and / or antibodies present in the reaction chamber from the photometric data; a control unit configured to execute a software in which the instructions to command the aforesaid means and units are coded according to the steps of the method. Also, at least one second reaction chamber identical to the first reaction chamber suitable for analyte analysis is provided and is subjected in succession or in parallel to said first reaction chamber to the steps of the method of analysis. Said second reaction chamber is provided in combination with feeding organs of a control analyte having known composition, in particular, in relation to antibodies and to allergens present in it; washing unit of said second reaction chamber for eliminating said control analyte and / or further reactants fed to said reaction chamber in successive times of analysis process; feeders of a further immuno-enzymatic reagent for example an enzyme conjugated to the specific antibody for the control analyte; feeding means of a detection reagent which determines an optically detectable modification of the reactants in the reaction chamber, in particular of the immuno-enzymatic reagent; and where said photometric sensor reads intensities of optical modifications of the immuno-enzymatic reagent upon reaction with the detector reagent in said second reaction chamber; while the processing unit is configured to receive the photometric measurement signals deriving from the analysis of the control analyte and determines, by interpolation with a reference curve, the quantitative numerical values of allergens and / or antibodies present in the reaction chamber from the photometric data, the said processing unit being further configured to compare the values obtained from the said interpolation with known values for the control analyte and to compare the difference between these values with at least one threshold and / or a minimum threshold for the said difference at least one generator of a commanded alarm signal, from said comparator when said difference is greater than the said maximum threshold or less than the said minimum threshold; and of an activation signal of a phase of recalibration or modification of the reference curve and / or curves for families of allergens and / or single allergens as a function of the differences detected between measured values and known values for the control analyte; said processing unit being further configured to recalculate the quantities values obtained for the examined analyte by interpolation of measurement signals of photometric sensor with the curve of modified or recalibrated reference curve.

According to an aspect of the invention, the system comprises at least one third reaction chamber dedicated to a verification analyte, which is identical to the reaction chambers provided for the examined analyte and for control analyte the third chamber being provided in combination with: feeders of a test analyte having a known composition, in particular, in relation to the antibodies and allergens present therein; washing unit of said second reaction chamber for eliminating said verification analyte and / or additional reagents fed to said reaction chamber of in successive times of the Process of analysis; feeders of a further immuno-enzymatic reactant for example an enzyme conjugated to the antibody specific for the verification analyte; feeders of a detection reagent which determines an optically detectable modification of the reactants in the reaction chamber, in particular of the immuno-enzymatic reagent. Said photometric sensor reads intensities of the optical modifications of the reagent immuno-enzymatic upon reaction with the detector reagent in said third reaction chamber. The processing unit is configured to receive the resulting photometric measurement signals from the analysis of the verification analyte and determines through numerical interpolation with the recalibrated or modified reference curve the quantitative values of the allergens and / or antibodies present in the reaction chamber from the photometric data, the said unit being further configured as a comparator to compare the values obtained from said interpolation with known values for the verification analyte and to compare the difference between these values with at least a maximum threshold and / or a threshold minimum for said difference and at least one generator of. an alarm signal controlled by said comparator when said difference is greater than said maximum threshold or lower than the said minimum threshold.

According to an aspect of the invention, the system comprises several reaction chambers are provided, preferably grouping in groups of five reaction chambers comprised in an integrated reaction device and at least some of the feeding means and / or washing units can be constituted by a general mean or can be mounted on a general movement system and / or connected to general systems for the distribution of solvents or washing liquids .

The system comprises at least one processing unit which is constituted by a processor 140 which executes a logic processing program 150 in which the instructions for configuring the processor to the processing of the data measured by the sensor 130 are coded, and in particular for determining by interpolation with a reference curve the quantities of allergens and / or antibodies present in the reaction chamber from the photometric data. A control unit 160 executes a logic control program 170 which includes instructions for configuring it to operate as a synchronized control unit of the aforesaid organs and the aforesaid units correspondingly to the steps of the analysis method previously described. The photometric sensor 130 is also intended to read the intensities of the optical modifications of the immuno-enzymatic reactant during the reaction with the detector reagent in the said second reaction chamber in which the reaction takes place with a reference analyte having a known composition. Thanks to the processing software 150 and to a memory 180 which includes at least the reference curves 181 and / or the known values of the composition of the control analytes 182, the processing unit 140 is configured to receive the photometric measurement signals deriving from the analysis of the control analyte having known composition and to determine by interpolation the numerical values deriving from the photometric data from or from the sensors 130. An input / output interface 190 with the user, can alternatively or in combination graphical interfaces, keyboards, mice, touch screens, monitors, acoustic interfaces, mobile units, tablets etc. Alternatively, or in combination, reading and / or writing units for portable storage media (USB memories, Cd, DVD-RAM etc) are also possible as interfaces. Through one or more interfaces it is possible both to modify the software processing settings 150 and logic control 170 and to load data 181, 182, 183, 184 and the programs 185, 186 in the memory 180. The user interfaces 190 also comprise at least one alarm signal generator controlled by the comparator. The logic processing program 150 further comprises the instructions for the processor 140 to configure the same to generate a signal to activate a phase of recalibration or modification of the reference curve and / or curves for families of individual allergens as a function of the differences detected between measured values and known values for the control analyte. Therefore, advantageously the system and method for solid phase analysis of biological samples for analytes present in a liquid solution, especially for in vitro diagnosis of allergies and autoimmune diseases, provide an optimized reaction chamber conformation.

Another advantage of the system and method for solid phase analysis of biological samples is that they allow to optimize a reaction vessel so as to make the production and application phases of the solid phase internal to each reaction chamber easy and effective.

Finally, the system and method for solid phase analysis of biological samples according to the invention is easily achievable and of limited cost with respect to the accessible advantages.

Finally, it is clear that the system and method for solid phase analysis of biological samples herein described and illustrated can be subject to modifications and variations without departing from the scope of the present invention, as defined in the attached claims.