JPH028270B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention relates to immunoassays, in particular to qualitative and quantitative determination of immune responses. For more details,
The present invention relates to the use of electro-optical imaging to quantify the extent of aggregation that occurs in immune response systems, and to the determination of the concentration of immune response components that cause aggregation in immune response systems. The invention also relates to a device for carrying out this method. Efficacy assays of drugs in biological fluids have received considerable attention in recent years, and therapeutic monitoring of drugs has gained considerable clinical significance and contributed to the development of new potency assay devices and methods. Potency assay devices using enzyme-labeled antigens, haptens, or antibodies have been used to measure substances in a variety of biological fluids. G.Brian Wisdom, “Enzyme−
Immunoassay”, Clinical Chemistry, vol. 22, 8.
No. 1976, pages 1243-55. These assay devices are commonly used for enzyme immunoassay, abbreviated as EIA, and enzyme-linked immunoassay, abbreviated as ELISA.
(linked immunoassay). Other methods generate hapten-protein conjugates,
Includes radio-immunoassay (RIA), which attaches a radioactive label to a specific location on a drug molecule. Alan Broughton and James E. Strong
“Radio immunoassay of Antibiotics and
Chemotherapeutic Agents”Clinical
Chemistry, Vol. 22, No. 6, 1976, pp. 726-32 and R. Cleeland et al. “Detection of Drugs of
Abuse by Radio Immunoassay：A
Summary of Published Data and Some New
Information”Clinical Chemistry, Volume 22, Issue 6,
1976, see pages 713-25. Before the development of various modern test methods, antigen/
The presence of antibody complexes was usually detected visually.
However, the results are often of limited clinical significance because the naked eye cannot detect or discern marginal but clinically meaningful responses. Microscopic examination also provides limited clinical information, as quantitative results cannot be obtained. Although EIA methods are generally specialized and sensitive identification and quantification methods for a wide range of substances, they have several drawbacks and limitations. These methods are generally RIA
It is less sensitive than the conventional method and is more susceptible to interference. moreover,
Determination of the terminal point (initial rate of enzymatic reaction) is more difficult than RIA methods, and all EIA methods except homogeneous EIA methods require separation of bound molecules from free labeled molecules. However, the number of separation methods that can be used is usually limited. RIA methods, on the other hand, require the production of specific antibodies and generation of appropriate radioactively labeled compounds. Due to the radiation hazards, the use of this method requires great care and skill, and radiation damage can even adversely affect the immunochemical reactivity of the labeled substance. Other known test methods include spin immunoassays. This involves mixing a known amount of antibody against the drug to be detected with a nitric oxide-labeled (spin-labeled) analogue of that drug, and adding to it a sample of biological fluid. The concentration of unknown test drug is determined by electron spin measurements. Simon L.Sharpe
“Quantitative Enzyme Immunoassay：Current
Status”Clinical Chemistry, Volume 22, Issue 6, 1976
See 2013, pp. 733-38. Still other testing methods include attaching antibodies to fluorescent compounds that emit light when excited with light at specific wavelengths;
It involves using opaque colloidal particles such as kaolin, carbon and charcoal particles, and animal blood components, typically red blood cells, and attaching antigens/antibodies thereto. In the immunoassay described above, the labeled component of the antigen/antibody reaction binds to its complementary position.
The amount bound depends on the concentration of other components. When one of these concentrations changes, the distribution of the labeled component changes with it between the bound and unbound portions. The nature of the label determines its distribution, and a calibration curve can be constructed by relating the concentration of the varying (unlabeled) component to the labeled component. This is currently accomplished by solid phase absorption in test tubes or plastic beads, centrifugation, repeated resuspension, and removal of bound or unbound moieties by washing. Although these methods yield curves representing the relationship between bonded and unbound moieties, they require tedious, cumbersome, and complex equipment. Additionally, EIA, RIA, and spin immunoassays introduce labeled substances into the reaction system that adversely affect the immunoreactivity of the non-immunoreactive reaction system by chemical or radioactive action that can be utilized as a label. usually required. Classic visual indicators of immunoreactivity do not require phase separation and removal of components from the immunoreactive system, nor do they require the introduction of labeling substances that interfere with or alter the immunoreactivity of the reaction system. This indicator is not subject to the drawbacks of EIA, RIA and spin immunoassay methods. Rather, they are based on a uniform redistribution of opaque particles from a non-agglomerated state to an agglomerated state. Since each particle acts as a label for many immunoreactive protein molecules, some labeled particles aggregate to form characteristic micro-agglomerated structures. This is observed qualitatively by visual inspection. However, this method is not useful for quantitative measurements and therefore has very limited clinical significance as well. There are also many patents related to various detection methods of agglutination reactions. U.S. Pat. No. 3,074,853 describes an immunological reaction by preparing and spreading a mixture of the subdivided solid and the two liquids to be tested for antigen/antibody reactions in test spots on a contrasting opaque surface of a solid. A method and apparatus for performing this are presented. The test strip scores are then visually inspected. No. 3,520,609 describes a method for detecting agglutination reactions that involves scanning a sample with a beam of energy source (eg, light or electrons). Substantial aggregation creates significant boundaries between the aggregated cells in the scanning area and the surrounding area, and these boundaries modulate the scanning beam. As the beam crosses the boundary, the rate of signal generated changes. A second signal is taken from the rate of change in the signal produced by the beam of light, integrated over a predetermined time period, and the integrated value is compared to a predetermined standard value corresponding to substantial aggregation to determine the reaction area. Determine whether agglomeration has occurred. US Pat. No. 3,819,271 describes a method and apparatus for measuring cell aggregation in a carrier liquid.
According to this method, a carrier liquid with cells suspended therein is placed in a container and a circular path is moved to direct a light beam through the suspension. The degree of cell aggregation is determined by the amount of light scattered upon passing through the suspension. U.S. Pat. No. 3,984,533 describes an electrophoretic method for detecting antigen/antibody reactions, and U.S. Pat. No. 3,990,851
No. 5, No. 1, No. 1, No. 1, No. 1, No. 1, No. 1, No. 1, No. 1, No. 1, No. 1, No. 1, No. 1, No. 1, No. 1, No. 1, No. 1, No. 1, 2006, 2013, and No. 2, No. 1, No. 1, No. 1, No. 1, No. 1, No. 1, No. 1, No. 1, No. 1, and No. 1, No. 1, No. 1, No. 1, No. 2, and No. 1, No. 1, No. 1, No. 1, No. 1, and No. 2, No. 1, No. 1, and No. 1, No. 1, No. 1, and No. 1, No. 1, No. 1, 2003, 2004, and describes a method and apparatus for measuring antigen/antibody reactions by transmitting laser light through a reaction mixture and measuring the forwardly scattered light. US Pat. No. 3,905,767 describes a method for qualitatively or quantitatively measuring antigens or antibodies by antigen/antibody reactions in which a precipitate is formed. This patent also measures the amount of light that is scattered when a light beam passes through a precipitate. The methods and devices of these and other patents all require detection of agglutination reactions by photometric means, such as light scattering measurements, turbidity measurements, or opacity measurements. These measurements therefore have clinically significant errors due to the color or inherent haze of the biological fluid being tested. In addition, U.S. Patent No.
In the method of all the above patents except No. 3819271,
Non-uniform distribution of reaction indicators introduces statistical errors in testing by automated equipment and makes interpretation by manual techniques difficult. Therefore, the detection of immune responses is simple and quick to perform and provides clinically meaningful information without the difficulties and limitations inherent in or associated with currently available immunoassays. The need for a test method for quantitative determination has been felt for a long time. It is therefore an object of the present invention to provide an immunoassay method and a device for carrying it out for the detection and quantification of immune reactions. Another object of the present invention is to provide an immunoassay that provides more clinically meaningful information than is provided by currently available immunoassays and devices. Yet another object of the present invention is to provide an immunological test and device that detects and quantifies immune responses more quickly and accurately than conventional methods and devices. Yet another object of the invention is to provide an immunoassay that is non-invasive and does not require separation of binding from non-binding portions. Yet another object of the present invention is to obtain an immunoassay method and device that can extract more information from labeled components. Yet another object of the present invention is to obtain a uniquely designed, disposable, flat reaction imaging cell in which the immune reaction takes place. Yet another object of the invention is to provide a case for retaining a reaction video cell within the device of the invention during video analysis of the reactants within the video cell. The present invention provides a unique device and novel method for detecting and quantifying immune response components of the immune response system. The device includes a retaining case formed by side walls and a front and back wall, and a plurality of spaced, axially aligned compartments between the side walls. Each compartment holds a uniquely designed video cell embodying the invention. Each video cell is made up of a pair of opposing, parallel, flat surfaces. Each of these surfaces has a roughly circular depression that, when joined together, creates a reaction cell. One of these circular depressions has a port through which the fluid sample and reactants are introduced into the reaction cell. The apparatus also includes a device, such as a caliper member, which is inserted into the compartment to raise the image cell part way into the optical path of the beam of radiant energy projected from the monochromatic light source. There is. The light beam transmitted through the image cell is transferred to a charge-coupled device (Charge-Coupled device) by an imaging lens.
Focus on the surface of an image detector such as a device. In this method, a biological fluid test sample and a reagent (e.g., substantially opaque colloidal particles such as latex spheres) are pipetted into a reaction cell through an input port, mixed uniformly, and the reaction mixture is incubated. (proceeding the reaction) to create aggregated particles,
Transilluminating the contents of the reaction cell. The transmitted light is imaged onto the surface of the image detector, creating a dark area corresponding to the aggregated particles in the reaction cell. The total dark image area is then measured, preferably with an electronic device. This procedure is repeated for test samples with known antibody concentrations and the resulting dark areas are plotted against these concentrations. The concentration of the immunoreactive component of the unknown test sample is then obtained from the plot. The present invention provides a unique method and apparatus for conducting immune reactions and qualitative and quantitative determination of the immune components of the reaction system. The unique method of the invention is characterized by the application of electro-optical imaging to a sample of the immune system and the cartographic analysis of the optical image produced on the imaging detector. The degree of agglutination in the immunoreaction medium is thus quantified and is related to the concentration of the immunoreaction components (ie, antigen and antibody) that cause the agglutination. An embodiment of the present invention will be described using the following figures. FIG. 1 shows a uniquely designed immunoreactive imaging cell 1 for use in the photovoltaic device of the present invention. The image cell 1 comprises two opposite parallel flat surfaces 3, 5, which are suitably glued together, preferably with an ultrasonic device, to form side edges 7, 9 and top and bottom edges 11, 13.
It shall be an integral structure defined by Sides 7 and 9 are slightly tapered and the lower end of side 9 is chamfered 15 to facilitate insertion of the video cells into their respective compartments 17 of holding case 19 in FIG. A pair of notches 2 are provided on the sides 7 and 9 of the video cell.
There are 1, 23, 25, 27, and the video cell is divided into 1
When inserted into section 7 , it is adapted to engage with an elastic projection 29 formed on the wall of compartment 17 or channel 31 . As shown in FIG. 3, the holding case 19 is made up of front and rear panels or walls 33, 35 and side panels or walls 37, 39. A plurality of generally groove (U) shaped partition walls 41 are spaced along the main axis of the holding case 19 and are suitably attached to the side walls 37, 39 to define respective compartments 17. The side walls 39 are partial walls extending to slightly more than half the height of each compartment. The lower end of the side wall 37 has a shoulder 43 inside, and when the video cell is fully inserted into the compartment, the ear 45 formed on the bottom side 13 of the video cell rests on the shoulder 43.
This arrangement and the engagement of the notches 21, 23, 25, 27 with the protrusions in the channel 31 prevent the video cells from slipping out of the compartment. Further, as shown in FIGS. 3 and 4, the holding case 19 has lateral flange members 47, 49 which are convenient for pinching it between the fingers and moving it into and out of the holding case of the device of the invention. be. Referring again to FIG. 1, each of the opposing surfaces of the two planes 3, 5 has a generally circular groove forming a reaction cell 51 in which the immune reaction takes place when the two surfaces are joined as described above. In the groove of the plane 3 there are holes 53 for introducing reagents and biological fluid into the reaction cell.
Open. A focusing target 55, for example a cylinder, is molded onto a round groove on the inner surface of the plane 5 of the reaction cell 51 so that the optical system is focused in the middle of the reaction cell. The two planes 3, 5 are made of glass or a suitable plastic material such as polystyrene. Both sides can be made of transparent material;
The other may be semitransparent. In the latter case, the plane 3 is made of a transparent material and the plane 5 is made of a translucent material. 3 and 4, in order to introduce reagents and biological fluid samples into the reaction cell, pinch the sides 7, 9 of the imaging cell between your fingers so that the notches 21, 25 are inside the channel 31. from the compartment until it engages the corresponding protrusion of the Reagents and biological fluid are then sequentially introduced into reaction cell 51 through hole 53 using a pipette or other suitable means. The image cell is then pushed fully into the compartment and the next image cell is filled with reagents and biological fluid as described above. The plurality of reaction cells are thus filled with reagents and the desired biological fluid to be tested. After the reagents and biological fluids are placed in the reaction cell as described above, the holding case is gently rotated about its axis of rotation to mix and evenly distribute the reagents and biological fluids. Figures 6a-6d show the respective positions of the video cells during rotation of the holding case. Since a so-called toroid-shaped meniscus is formed around the rotation hole 53 (FIG. 7), no fluid can escape from the reaction cell 51. Although the apparatus of the present invention is designed to self-rotate until the desired degree of mixing and uniformity is achieved,
The holding device 19 may also be rotated by hand. However, the rotational speed of the holding case may be varied as long as the centrifugal force of the fluid in each reaction cell does not exceed the surface tension of the liquid meniscus. As a practical matter, gently rotating the holding case for several minutes is adequate for proper mixing and uniform distribution of the reactants in the reaction cell. Before the imaging operation described below, the contents of the reaction cell are cultured under appropriate culture conditions (the reaction is allowed to proceed). Culture conditions vary depending on the reagents and biological fluid. Generally, the temperature of the fluid in the reaction cell during cultivation is maintained at about 25°C to 50°C for about 5 minutes to 15 minutes. Incubation is carried out with the imaging cell in a holding case in a suitable chamber of the device of the invention, or, if preferred, in a separate chamber. Reagents suitable for the practice of this invention are approximately 1 micron-
Virtually opaque colloidal materials such as charcoal, latex spheres, glass or ceramic spheres, kaolin earth, polystyrene beads, mammalian red blood cells, etc., with a particle size of 5 microns or somewhat larger. Although latex and charcoal particles are preferred due to their high opacity and easy availability, they are generally used as reagents in the practice of this invention. In FIGS. 8 and 9, the holding case 19 is placed on a conveyor belt 57 that is wrapped around a pulley 59 driven by a motor 61. In FIGS. Conveyor belt 57 has lug 6
3 and engages with the protrusion 65 of the holding case when the holding case is sequentially advanced to a predetermined position after each image operation. To lift the image cell out of the compartment and into the optical path, the apparatus of the invention includes an upper caliper section 69.
There is a caliper member 67 including a lower caliper portion 71 and a lower caliper portion 71 . The lower caliper portion 71 has an integral vertically projecting arm 73 which is inserted into a compartment of the holding case to lift the image cell into the optical path. The lower caliper section has a lateral section 75 fixed to a belt 77 which is placed around a pulley 79 driven by a motor 80. The upper and lower caliper parts have guide rods 81
It is usually fixed to a spring load member 83 including spring members 85 and 87 and biased. The upper caliper portion 69 is maintained above the image cell by a stop 89 on the guide rod. In order to bring the video cell into the optical path, the motor 80 is driven by a switch (not shown) to move the belt 77 vertically upward. In this way, the vertically protruding arm 73 of the lower caliper part engages the lower side of the video cell, and as the belt continues to move, the video cell is pushed up and pressed against the upper caliper part 69, holding it as shown in FIG. Partially push it out of the case and put it in the optical path. After image processing of the contents of the reaction cell, the belt 77
The direction of movement is reversed to return the video cell to its original position within the compartment. Thereafter, the motor 61 is driven to move the belt 57 in the direction of the arrow in FIG. When the lug 63 engages the next protrusion of the holding case corresponding to the next compartment, the motor 61 is automatically turned off and the motor 80 is turned off.
is driven to bring the next image cell into the optical path for image processing. This operation is continued to process the video of a desired number of video cells. As shown in FIGS. 10 and 11, the electro-optical arrangement of the apparatus of the present invention includes a radiant energy source, such as a monochromatic light source 93. As shown in FIGS. A monochromatic light beam from a light source 93 enters the image cell 1 and transilluminates the contents 52 of the reaction cell 51 . The area transilluminated by the light beam contains a polydisperse uniform distribution of labeled latex particles representing the total reaction medium within the reaction cell. The depth of the reaction cell 51 must be minimal in order to concentrate a large proportion of the substantially opaque colloidal particles in the focal plane of the optical system, which is aligned with the image plane from which all information is extracted. The light beam emitted from the light source 93 is made into a parallel beam and collected by a condenser lens (not shown).
The light transmitted through the reaction cell 51 is passed through an imaging lens 95 of an appropriate diopter. The imaging lens 95 has rack and pinion devices 99, 101 for adjusting the imaging lens.
and is mounted within an imaging device 97 operatively connected by. The light beam transmitted through the imaging lens 95 is incident on the surface of the image detector 103 to form an image of the aggregated substantially opaque colloidal particles. The image detector 103 is a Charge-Coupled Device, commercially abbreviated as CCD, and is a charge-coupled device commercially abbreviated as CCD.
Device”, Scientific American, February 1974,
It may be of the type described on pages 23-33. CCD is
It includes a silicon grid made of 10,000 photosensitive elements arranged in a 100 x 100 square and metal contacts around the periphery of the grid for connection to metal connectors. When light is incident on the surface of a silicon grid, it is absorbed and electrons are liberated in an amount proportional to the amount of incident light. Thus, in accordance with the present invention, substantially opaque colloidal particles in the biological fluid within reaction cell 51 are transilluminated and imaged onto the photosensitive surface of the CCD. The magnification of the optical system is adjusted so that the size of the image of non-agglomerated particles is smaller than the area of each photosensitive element (image element or pixel). Since the electrical output from the photosensitive elements is proportional to the amount of light incident on their surface, if the unagglomerated particle image is smaller than one pixel, the electrical output is less than the electrical output corresponding to a completely shaded pixel. Therefore, all images whose electrical output is less than a predetermined threshold level, such as non-agglomerated particles, i.e. particles that are close to each other but not in any contact, allowing some light to pass through. Limit rejection criteria can be determined to discard areas. When several substantially opaque colloidal particles aggregate into a large mass, the image appears in the first
As shown in Figure 2, aggregated particles that cover several pixels are tightly packed together, so they appear darker than a single particle. After imaging the aggregated particles as described above, the CCD is electronically scanned row by row. Pixels that fall into portions of the image that are darker than the threshold level generate an electrical (voltage) output that is a function of the image density of aggregated particles. FIG. 13 shows the electrical output corresponding to the agglomerated grains on the photosensitive surface of the CCD of FIG. The method of the invention is particularly suitable for determining disease states such as syphilis, hepatitis, etc. Generally, the method involves coating substantially opaque colloidal particles (latex particles) with disease-associated antigens. Antigens are attached to the surface of the latex by absorption, adsorption, chelation, or other conventional techniques known in the art. A compatible immune response component is present in the analyte, ie, the biological test sample, such as serum. When the analyte is added to the latex-coated particles, it is believed that the reaction between the antigen and antibody creates a physicochemical complex that causes aggregation of the latex particles. The degree of aggregation depends on the number of matched positions, and the reaction proceeds until all antigen positions are bound by antibodies in the analyte. In this way, clumps, ie agglomerated particles of different dimensions, are created in the reaction medium. Their number and size are proportional to the relative concentrations of antigen/antibody and substantially opaque colloid particles in the analyte. After imaging the aggregated particles by transilluminating the reaction cell, the image area (image area) is quantified (electronically) to obtain the total area. This is a function of the concentration of antibody in the analyte. The above method can be used for qualitative determination of unknown samples. The above method is repeated for the reference sample and the total dark image area is compared to determine the nature of the unknown sample. To determine the concentration of the immune component in an unknown biological sample, the above method is repeated on at least two control samples of known but different antibody concentrations. One of the control samples is a negative control (serum without antibodies). The total dark image area of the control sample is then related to each concentration. The antibody concentration of the unknown sample is obtained from the total dark image area obtained for the unknown sample and the relationship between the total dark image area of the control sample and its concentration. The photoelectric device of the present invention will be explained using the block diagram shown in FIG. Reaction cell 5 as shown
1 is transilluminated by a monochromatic light beam from a light source 93. The light beam transmitted through the image cell is sent to the imaging lens 95.
Focus on CD103. Divide the CCD output into two parts. Meanwhile, 105 supplies averaged data to an intensity feedback controller 106. the other 1
07 provides data for determination of granularity and frequency of occurrence. A second path 107 leads to a threshold comparator and particle counter 109 that excludes unagglomerated particles from both intensity and size. Comparator 109 passes through tightly packed particles (i.e., agglomerated particles) and excludes counts of out-of-focus particles, which produce a very dark image on CCD 103 versus a faint image of non-agglomerated particles. A display cathode ray tube 111 assembles the sequential output data from the CCD 103 into an image of the reaction field and displays it as an image of the test sample, providing visual feedback for determining system operation. Temperature control device 113 monitors and controls the temperature of the contents of the video cell to a predetermined optimum level. The apparatus shown in FIG. 14 also includes a digital computer 115.
and a control logic unit 117 that monitors and controls the main functions of the device. The storage device 119 stores the output of the calculation, and the printing device 121 prints the output of the computer. Next, two examples of the present invention will be shown. It should be noted, however, that these examples do not limit the scope of the invention or its applicability to other disease states. EXAMPLE This example demonstrates the applicability of the method and apparatus of the invention to syphilis. Rapid Reagent (RPR) is a government-supplied reagent manufactured by the Venereal Disease Reagent Laboratory.
Plasma Reagent) Charcoal Antigen Suspension (HWD)
60 microliters were pipetted into each of the 30 video cells placed in the three holding cases described above. Additionally, 40 microliters of the control serum described below (obtained from a syphilis patient) was placed in each of cells 1-4.

[Table] An additional 40 microliters of suspected patient serum was added to each of the remaining 26 image cells, and these cells were incubated by gently rotating the holding case at a temperature of 30° C. for 8 minutes. The degree of aggregation in each cell was then determined using the method and apparatus described above. The results are shown in the table.

【table】

Table As shown in the table, various scores can be compared to control samples to determine the extent of disease in the test sample. For example, the fifth cell indicates a weak response state and therefore a weak disease state. Cells 6-10 indicate no syphilis. Cell 11 indicates moderate disease status, and cells 20 and 28 indicate advanced disease status. Furthermore, the results obtained can be quantified in percent of the positive control as follows. Machine Score/1500 x 100 For example, a machine score of 998 in cell number 10 means that the suspect sample concentration is 998/1500 x 100 or 66.53 percent of the positive control in cell number 1. Thus, once the concentration of the positive control is known, the degree of disease state of the test sample can be determined by the method and apparatus of the present invention. Example This is human chorionic gonadotropin.
This is an example of determining pregnancy by testing for Chorionic Gonadotropin (HCG). Seventy-five microliters of anti-HCG/latex bead reagent suspension (HWD) was placed into ten imaging cells loaded into holding cases. Two cells were added with 25 microliters of urine and used as a control. One is known reactivity and the other is unknown reactivity.
The remaining image cells contained 25 microliters of test sample. The holding cases were incubated at a temperature of 28°C for 8 minutes with gentle rotation and evaluated using the apparatus and methods described herein. The results are shown in the table.

【table】

Table 1 It will be appreciated that many variations can be made to the method and apparatus of the invention described above and are within the scope of the invention.

[Brief explanation of drawings]

FIG. 1 is a perspective view of a reaction video cell used in the practice of the present invention, FIG. 2 is a sectional view taken along line 2-2 in FIG. 1, and FIG. 3 is a perspective view of a holding case for the video cell of the present invention. , FIG. 4 is a cross-sectional view taken along line 4--4 in FIG. 3, FIG. 5 is a cross-sectional view taken along line 5--5 in FIG.
Figures 6a-6d are diagrams showing the positions of the reaction cells in the image cell being rotated about the central axis to achieve mixing and uniform distribution of the reactants; Figure 7 is the illustration of the reaction image cell in rotation; A cross-sectional view taken along line 7--7 of FIG. 6a showing the toroidal meniscus created in the filling hole; FIG. FIG. 9 shows the device of the invention to the extent that it shows both lateral displacement and
The figures are cross-sectional views taken along line 9--9 of FIG.
Figure 11 is a diagram showing the process of creating an image of aggregated labeled particles in a suspension in a reaction cell, Figure 12 is a front view of the image detector where an image of aggregated particles is created, and Figure 13 is the image detector. FIG. 14 is a graph of the electronic signal corresponding to the above agglomeration region, and is an assembled diagram of the optoelectronic device used to implement the present invention. 1...Video cell, 19...Holding case, 51...
... Reaction cell, 53 ... Filling hole, 55 ... Focusing target, 61 ... Motor, 67 ... Caliper, 80 ... Motor, 81 ... Guide rod,
93...Light source, 103...Video detector.

Claims (1)

[Scope of Claims] 1. A method for the rapid and accurate qualitative determination of immunoreactive components of a biological fluid test material (sample), comprising: (a) treating said specimen with antigen in substantially opaque colloidal particles; (b) uniformly mixing and incubating the analyte and the reagent in the reaction area; and (c) irradiating the contents of the reaction area. (d) transilluminating with energy and imaging the transmitted light beam onto a surface of an image detector to form a dark area corresponding to the aggregated particles in said reaction region; (e) repeating steps (a)-(d) on the reference material; (f) measuring the area of the total dark image region of the specimen and the reference material; and comparing the area of the total dark image region of the document. 2. The method of claim 1, wherein the reagent is charcoal particles. 3. The method according to claim 1, wherein the reagent is a latex sphere. 4. The method of claim 1, wherein the culturing is carried out at a temperature of about 25°C to 50°C for about 5 minutes to 15 minutes. 5. The method according to claim 2, wherein the culturing is carried out at a temperature of about 25°C to 50°C for about 5 minutes to 15 minutes. 6. The method of claim 3, wherein the culturing is carried out at a temperature of about 25°C to 50°C for about 5 minutes to 15 minutes. 7. The method according to claim 1, wherein the area of the region corresponding to the aggregated particles is measured by an electronic device. 8. The method according to claim 2, wherein the area of the region corresponding to the aggregated particles is measured by an electronic device. 9. The method according to claim 3, wherein the area of the region corresponding to the aggregated particles is measured by an electronic device. 10. The method according to claim 4, wherein the area of the region corresponding to the aggregated particles is measured by an electronic device. 11. The method according to claim 5, wherein the area of the region corresponding to the aggregated particles is measured by an electronic device. 12. The method according to claim 6, wherein the area of the region corresponding to the aggregated particles is measured by an electronic device. 13 A method for rapidly and accurately determining the concentration of an immunoreactive component in a biological fluid test material (specimen), which method comprises: (a) converting the specimen into a specimen reagent in which substantially opaque colloidal particles are coated with an antigen; (b) uniformly mixing and incubating the analyte and the reagent in the reaction zone; and (c) transilluminating the contents of the reaction zone with radiant energy. and (d) imaging the transmitted light beam onto the surface of an image detector to form a dark area corresponding to the aggregated particles in the reaction region; and (d) total darkness on the surface of the image detector. (e) repeating said steps (a)-(d) for at least two control samples of known but different antibody concentrations; and (f) measuring the area of said control sample. (g) determining the antibody concentration of said specimen from the relationship obtained from said step (f). 14. The method according to claim 13, comprising:
A method in which the reagent is charcoal. 15. The method according to claim 13,
A method in which the reagent is a latex sphere. 16. The method according to claim 13, comprising:
The incubation is carried out for about 5 minutes at a temperature of about 25°C - 50°C.
How to do it for 15 minutes. 17. The method according to claim 14,
The incubation is carried out for about 5 minutes at a temperature of about 25°C - 50°C.
How to do it for 15 minutes. 18 The method according to claim 15, comprising:
The incubation is carried out for about 5 minutes at a temperature of about 25°C - 50°C.
How to do it for 15 minutes. 19 The method according to claim 13,
A method in which the area of the region corresponding to the aggregated particles is measured using an electronic device. 20 The method according to claim 14,
A method in which the area of the region corresponding to the aggregated particles is measured using an electronic device. 21. The method according to claim 15,
A method in which the area of the region corresponding to the aggregated particles is measured using an electronic device. 22 The method according to claim 16,
The area of the region corresponding to the aggregated particles is measured using an electronic device. 23 The method according to claim 17,
A method in which the area of the region corresponding to the aggregated particles is measured using an electronic device. 24 The method according to claim 18,
A method in which the area of the region corresponding to the aggregated particles is measured using an electronic device. 25 (a) a retaining case formed by front and rear walls and side walls with a plurality of spaced axially arranged compartments between said side walls; (b) formed by two opposed parallel planes; , each said surface has a generally circular groove, and when said surfaces are joined together, the front circular groove forms a reaction cell, and said groove has a biological fluid (analyte) and an analyte in said reaction cell. (c) a device that engages and biases each said image cell to partially push it out of its respective compartment and into the optical path; (d) a light source that transmits a light beam to the reaction cell; (e) an image detector having a photosensitive surface made of a photosensitive element; and (f) a light source that transmits the light beam transmitted through the reaction cell to the image detector. (g) a device for measuring the area of the total dark image region on the surface of the image detector; Also, devices used for immune reactions. 26 The device according to claim 25,
The retaining case includes an upper open bottomless chamber formed by front and rear walls and a pair of opposing spaced side walls, and a plurality of spaced axial, generally rectangular compartments of the side walls. , a first lateral flange attached to one of the side walls, and a second lateral flange attached to the other of the side walls. 27. The device of claim 25 or 26, wherein one of the side walls of the holding case is a partial wall. 28. The device of any one of claims 25 to 27, wherein the holding case includes at least two compartments. 29. The device according to any one of claims 25 to 28, wherein both surfaces of the video cell are made of a transparent material. 30. The device according to any one of claims 25 to 28, wherein one of the surfaces of the video cell is made of a transparent material and the other surface is made of a translucent material. . 31. The apparatus of claims 25 to 30, wherein the imaging cell is arranged on the inner surface of one of the circular grooves such that the optical system focuses on an intermediate surface within the reaction cell. A device that has a cylindrical member that you want to attach. 32 The device according to claim 25, which
The device that engages and biases the image cell is a caliper device. 33. The device according to claim 26,
The device that engages and biases the video cell is a caliper device. 34. The device according to claim 27,
The device that engages and biases the video cell is a caliper device. 35 The device according to claim 28,
The device that engages and biases the video cell is a caliper device. 36 The device according to claim 29,
The device that engages and biases the video cell is a caliper device. 37 The device according to claim 30, which
The device that engages and biases the video cell is a caliper device. 38 The device according to claim 31,
The device that engages and biases the video cell is a caliper device. 39 The device according to claim 25,
The device for measuring the area of the total dark image area on the surface of the image detector is an electronic device. 40 The device according to claim 26,
The device for measuring the area of the total dark image area on the surface of the image detector is an electronic device. 41 The device according to claim 27,
The device for measuring the area of the total dark image area on the surface of the image detector is an electronic device. 42 The device according to claim 28,
The device for measuring the area of the total dark image area on the surface of the image detector is an electronic device. 43 The device according to claim 29,
The device for measuring the area of the total dark image area on the surface of the image detector is an electronic device. 44 The device according to claim 30,
The device for measuring the area of the total dark image area on the surface of the image detector is an electronic device. 45 The device according to claim 31, which
The device for measuring the area of the total dark image area on the surface of the image detector is an electronic device. 46 The device according to claim 32,
The device for measuring the area of the total dark image area on the surface of the image detector is an electronic device. 47 The device according to claim 33, which
The device for measuring the area of the total dark image area on the surface of the image detector is an electronic device. 48 The device according to claim 34, which
The device for measuring the area of the total dark region image area on the surface of the image detector is an electronic device. 49 The device according to claim 35,
The device for measuring the area of the total dark image area on the surface of the image detector is an electronic device. 50 The device according to claim 36,
The device for measuring the area of the total dark image area on the surface of the image detector is an electronic device. 51 The device according to claim 37,
The device for measuring the area of the total dark image area on the surface of the image detector is an electronic device. 52 The device according to claim 38,
The device for measuring the area of the total dark image area on the surface of the image detector is an electronic device. 53 (a) an imaging cell containing a biological fluid material (sample) and reagents for the specimen; (b) a light source projecting a beam of radiant energy onto said imaging cell; and (c) a photosensitive surface made of a photosensitive element. (d) an imaging lens for focusing light transmitted through the image cell onto a surface of the image detector to create a dark image area corresponding to aggregated particles within the image cell; (e) a threshold comparator for counting the area of dark image areas larger than a predetermined threshold value excluding dark image areas smaller than a predetermined threshold value; Device. 54. The photoelectric device according to claim 53, comprising a temperature control device for controlling the temperature of the contents of the video cell. 55. The photoelectric device according to claim 53, which visually displays the contents of the video cell.
A photoelectric device comprising a display screen operatively associated with said image detector. 56. A photoelectric device according to claim 54, which visually displays the contents of the video cell.
A photoelectric device including a display screen operatively associated with the image detector. 57. The photoelectric device according to claim 53, which includes a digital computer linked with the threshold comparator, a device for storing the output of the computer, and a device for printing the output of the computer. Device. 58 The photoelectric device according to claim 54, which includes a digital computer linked with the threshold comparator, a device for storing the output of the computer, and a device for printing the output of the computer. Device. 59 The photoelectric device according to claim 55, which includes a digital computer linked to the threshold comparator, a device for storing the output of the computer, and a device for printing the output of the computer. Device. 60 The photoelectric device according to claim 56, which includes a digital computer linked to the threshold comparator, a device for storing the output of the computer, and a device for printing the output of the computer. Device.