WO1989001156A1 - Procede et dispositif d'examen de sang complet, d'autres fluides corporels et d'autres fluides biologiques - Google Patents

Procede et dispositif d'examen de sang complet, d'autres fluides corporels et d'autres fluides biologiques Download PDF

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Publication number
WO1989001156A1
WO1989001156A1 PCT/EP1988/000693 EP8800693W WO8901156A1 WO 1989001156 A1 WO1989001156 A1 WO 1989001156A1 EP 8800693 W EP8800693 W EP 8800693W WO 8901156 A1 WO8901156 A1 WO 8901156A1
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WIPO (PCT)
Prior art keywords
carrier
antigen
holder
blood
substances
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Application number
PCT/EP1988/000693
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German (de)
English (en)
Inventor
Olaf Thraenhart
Original Assignee
Seeger, Wolfgang
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Publication date
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Publication of WO1989001156A1 publication Critical patent/WO1989001156A1/fr

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Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
    • G01N33/56983—Viruses
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G01N33/54306—Solid-phase reaction mechanisms
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G01N33/54366—Apparatus specially adapted for solid-phase testing

Definitions

  • the invention relates to a method for examining whole blood, other body fluids and other biological fluids, and a device for
  • Antibodies and antigens in hormone preparations, in the urine or in sera that have been diluted to reduce the influence of other disturbing factors are examined.
  • Samples from several patients must come together so that a test can be carried out profitably. Carrying out the examination also requires specialist knowledge and the availability of special equipment which, since they are expensive, can only be used expediently in special laboratories. Samples must therefore first be packed and sent to an institute by post, unpacked, registered and handed over to the technical assistants. There, an examination protocol is to be completed from samples from several patients, so that the individual samples can be tested against various antigens or antibodies in several examination approaches. A protocol to test assessment again to create and provide the findings back to the post office 'then. This results in a period of time from the taking of the sample to the receipt of the finding of at least one, but usually several days.
  • the finding is only available at a time when the result can often only confirm the therapeutic measures that are immediately necessary when samples are taken in the best case.
  • No result is available in acute emergencies.
  • An example is neurological diseases such as herpes infections, in which the therapeutic use of an anti-herpes agent should be serologically secured.
  • values of accident victims who are considered organ donors are available at the earliest hours after the transplantation, although it can be vital for the patient to know whether the accident victim is infected, for example, with the AIDS virus.
  • the global spread of HIV infection like the diagnosis of hepatitis B virus infection, requires mass examinations. Such mass examinations will only be manageable and financially if it is possible to develop methods that can also be carried out by trained laymen outside of special laboratories. The examinations should also be possible in the medical practice or clinic.
  • test kits currently available for the investigation of antibodies and antigens in the ELISA are in principle based on the adsorption of antigens or antibodies on a transparent plastic surface or on corresponding spheres, which are placed in transparent test tubes. This is followed by further binding with an antibody or antigen in the form of an antigen-antibody reaction. Finally, an enzyme-labeled protein is added that reacts specifically with the previous reaction product. The amount of bound antibody or antigen in the unknown sample is visible as a color reaction after the addition of a substrate in the liquid phase and is measured visually or photometrically. _
  • Nitrocellulose paper has also been described by Pappas et al (J. Immunol. Methods, 64, 205, 214 (1983)), Walsh et al. (J. Immunol. Methods, 66, 99 (1984)) and Heberling, and Kalter (Develop. Biol. Standard. 64, 199 (1986)) and Schmeer (personal communication, 1986). Herbrink et al. (J. Immunol. Methods, 48, 293 (1982)) compared nitrocellulose and diazobenzyloxymethyl paper for the use of an antibody test. Giallongo et al. (J. Immunol. Methods, 52, 379 (1982)) and Esen et al. (Analytical Biochemistry, 132, 462 (1983)) use chromatography paper as a carrier for protein determinations.
  • the object of the present invention is to provide a method for the examination of whole blood, other body fluids and other biological fluids, which can be carried out particularly easily, quickly and with minimal expenditure on devices.
  • This object is achieved in that the liquid is mixed with a surface-active medium and then subjected to test reactions.
  • the invention first has the advantage that solid components such as cell components of the biological fluid to be examined are lysed or destroyed. This is particularly important in all such examinations in which a reaction on or at one location of a reagent carrier can be used. If solids, for example cell components of the blood, are deposited precisely at this point, they can prevent a reaction of this reagent with the reagent present in the biological body fluid. This risk is eliminated by the present invention.
  • this method has the great advantage that no devices such as centrifuges are required to prepare the blood.
  • it has the advantage that the biological liquid can be subjected to the test reactions in a very short time after its removal, namely immediately after mixing with a surface or surface-active medium.
  • the biological fluid can be diluted with a surfactant.
  • the medium is a surfactant. It can be a surfactant NP 40 in a concentration of about 0.1%.
  • the test reaction for determining proteins with antibody or antigen-specific properties comprises the following steps:
  • the invention comprises the test reaction for the determination of antibodies or antigens the following steps: fixation of one or more first substances with antigen-specific properties on a support, supply of a second substance mixture, the antibody-specific components to be examined with the first substances during a first incubation, can undergo specific test reactions, to the first fabrics; Application of a first indicator which is uniform for the different reactions and which reacts with components of the second substance mixture attached to the first substances during a second incubation period and thus indicates the specific reaction which may have taken place.
  • the first advantage is that a small amount of blood is sufficient. This has the further advantage that no disposable syringe is required to draw blood. It also eliminates the difficult process of taking blood from the vein.
  • centrifuge that is otherwise required is not required. This means that the method according to the invention can be carried out anywhere, not only in laboratories which are equipped with centrifuges and other devices which are otherwise necessary.
  • Confusion risks such as those that occur again and again in the above procedure, are excluded in the examination according to the invention, since this begins in the presence of the patient.
  • the time from taking blood to receiving the findings is reduced from days to less than an hour.
  • the finding is still available during the first examination of the patient and can therefore be used therapeutically immediately.
  • the detection method according to the invention thus fulfills all the requirements for immediate emergency diagnostics at the bedside or even in the ambulance.
  • Serological examinations in an emergency are already of importance in brain inflammation caused by herpes virus infections and in organ transplants when organs are taken from accident victims and used in other patients.
  • Serological emergency diagnostics will be required with the expected development of chemotherapeutic agents against viral infections in the near future.
  • the present method for the determination of antibodies or antigens can also be carried out by the layperson, whereby the costs of the method are considerably reduced.
  • Another advantage can be seen in the fact that with the present method, because it is so easy and inexpensive to carry out, serial examinations of entire groups of people are also possible for the first time. Furthermore, the invention allows allergy examinations, which were previously very complex.
  • the present method can also be used to test vaccines for their antigen content before they are used. This is particularly important in the tropics, since the cold chain, which is often interrupted there, makes it easy for the vaccines to lose their active ingredient.
  • tropical countries and vaccines after their manufacture or import u. U. Store for a long time and then do not use later because of the uncertainty of the residual activity. In this way, large resources are wasted. The same applies to immunoglobulin preparations.
  • capillary blood is used. This further development has the great advantage that the blood does not have to be taken with a syringe, which the layperson is generally unable to do. Rather, it is possible to draw capillary blood with a lancet.
  • Fingertip or the earlobe This process is very simple and can be carried out by anyone.
  • Antibodies and / or antigens in drops of blood, capillary or venous blood are examined directly.
  • blood had to be centrifuged beforehand so that the corpuscular components interfere with the reaction.
  • the disturbing influence is eliminated by mixing according to the invention with a special buffer.
  • the disturbing influences in blood drops, capillary or vein blood can be eliminated by mixing with a medium.
  • the capillary blood can be mixed with the medium in a ratio of at least 1: 2 or 1: 3, preferably 1:50 to 1: 100.
  • the medium can contain a surfactant and a protein.
  • the surfactant can be NP 40. According to one embodiment of the invention, the concentration of NP 40 is about 0.1%.
  • the protein can be fetal calf serum, the concentration of which can be, for example, 3%.
  • antibodies and / or antigens in body fluids such as urine, cerebrospinal fluid or saliva can be examined directly.
  • Antibodies or antigens can also be detected in body fluids other than blood. Since HIV also occurs in saliva, for example, tests can also be carried out on these samples. The invention thus has the further considerable advantage that the blood can be dispensed with if the detection sensitivity is sufficient. Antibodies and / or antigens can be examined in prepared body fluids such as blood serum, blood plasma, or others. Antibodies can also be tested in immunoglobulin preparations.
  • Another important advantage of the present invention is that one or more antigens are tested in vaccines.
  • Enzyme-linked species-specific anti-immunoglobulins of classes G, M, A, D, E and their subclasses can be used as the first indicator.
  • antiimmunoglobulins of the different classes or subclasses are then to be used.
  • an examination with anti-immunoglobulin-A is carried out.
  • the first indicator can contain one or more enzyme-coupled antigen-specific antisera.
  • Peroxidase can be used for enzyme coupling.
  • the mixture is poured off after the incubation. This shortens the time required to carry out the method, since pouring can be carried out considerably faster than a washing process.
  • the carrier is advantageously washed.
  • the antigens directed against the antibodies are applied crude, partially or highly purified or as recombinant products or as antigen-containing or non-antigen-containing cells or as cell preparations on a non-soluble carrier from which the antigens be absorbed or on which they dry up.
  • the carriers can be materials which, owing to the contrasting color, allow a color reaction to be read directly by the eye. This enables examinations on simple papers (for example typewriter paper). The paper must have contrast (white) and should not be too absorbent in order to optimally design the distribution density of the first material in the carrier / surface unit.
  • Nitrocellulose e.g. Nitrocellulose, Chromatographie ⁇ papers or filter papers can be used.
  • Transparent materials and plastics can also be used as supports.
  • between 1 and 8 al of the first substance can be dropped onto the support.
  • about 1 drop of the first substance is dropped from a cannula onto the carrier.
  • the first substance can be dried briefly at a temperature of more than + 20 ° C. A drying temperature of around 37 ° C is suitable.
  • the drying time can be 10-15 minutes, depending on the drying temperature.
  • the supports are coated in the dry state and are used directly after drying or fixing the antigen or antibody.
  • Phosphate-buffered salt solution with a surfactant and a protein additive for example NP40 in a concentration of about 0.1%, can be used as the dilution buffer for the second substance mixture and / or the uniform indicator.
  • Fetal calf serum in a concentration of 3% can be used as a protein additive.
  • the buffer for the washing operations which may be carried out can be a surfactant in phosphate-buffered saline, preferably 0.1% NP40.
  • the process is advantageously carried out at a temperature between 1 and 100 ° C.
  • the temperature erhschreibung of 20 ° C to 37 ° C can shorten the reaction time to 'the order of 40-50%, depending on conditions Bedin ⁇ even stronger.
  • an antibody or an immune class-specific standard is examined simultaneously for the semi-quantitative determination of the antibody content, and the color reaction of the standard is compared with the result of the patient sample.
  • a carrier is used on which several first substances with antigen-specific properties are or are fixed; furthermore, it is not a second substance but one second substance mixture, the antibody-specific constituents to be investigated which can undergo specific test reactions with the first substances during a first incubation, is brought together with the first substances, and a uniform first indicator is applied for the different reactions, which is used during a second incubation period reacts with constituents of the second mixture of substances attached to first substances, and thus indicates the specific reaction that may have taken place.
  • This development of the invention makes it possible to use a single drop of blood to carry out a large number of antibody and antigen tests in a test batch in a group- or disease-associated manner, by applying a plurality of antigens and / or antibodies to the carrier.
  • the selection of the antigens and / or antibodies per test batch depends on the requirements that must be met for a screening of specific groups of people, patients and animals.
  • the selection of the antigens or antibodies can also be made according to those for the diagnosis of a clinical picture, a group of symptoms or organ disease.
  • Examples of group or disease-related examinations are: a) Blood donors at risk
  • antibodies or antigens of one or more pathogens are simultaneously detected which are relevant for the diagnosis of a symptom picture, an organ or systemic disease (disease-associated determination).
  • routine examinations which must always be carried out in the same way for certain groups of people, and which cover a specific spectrum
  • Antigen-antibody determinations include, can be carried out simplified (target group-associated determination).
  • a major advantage in establishing such a method is the method of testing for a large number of antibodies with a single drop of blood, which is presented here for the first time. Capillary blood can also be used, which is taken in the course of other routine examinations without severe patient nuisance.
  • This blood collection is also so simple that it can be carried out by trained laypersons. It is also an object of the present invention to provide a device for the detection of antibodies and / or antigens which is simple to produce, economical and easy to use.
  • this object is achieved by a body with a recess which is adapted in shape to accommodate one or more carriers to which one or more first substances with antigen-specific properties are fixed.
  • the recess can be covered in a water-tight manner and is dimensioned such that its contents can be mixed by shaking the body.
  • Such a body can be constructed and designed in a very simple manner.
  • the distance from the surface or surfaces of the carrier can be approximately one and a half times as high or higher than the layer height of the solution to be brought together with the carrier.
  • FIG. 14 is a plan view of an examination device according to the invention, partially in
  • FIG. 16 is a top view of the device of FIG. 9,
  • 17 is a plan view of an inventive holder with an inserted carrier
  • Fig. 18 is a sectional view of a container with an inserted carrier holder.
  • Example 1 Chessboard titration of three sera for antibodies against rabies virus antigen.
  • FIG. 1 a shows a graphic representation of the test result with a serum with antibodies against rabies virus.
  • 1 shows 5 strips of nitrocellulose paper, each of which is divided into 6 fields.
  • 2 / al of a concentrated and purified inactivated rabies antigen with 680IU / ml in dilutions of 1:50 to 1: 1200 were dripped into these fields.
  • the 1: 200 dilution of antigen thus contained 0.007 IU / 2 / ul.
  • 4 ml of the serum dilutions from 1:50 to 1: 600 were each incubated with a strip for 10 minutes at + 37 ° C with gentle shaking.
  • the liquid was then suctioned off and the strips were washed by placing them in about 5 ml of washing buffer for 5 minutes at + 37 ° C. with gentle agitation. This process was repeated 3 times.
  • the last washing buffer is aspirated thoroughly and then 4 ml of an anti-human IgG (conjugate) coupled to peroxidase are incubated in a dilution of 1: 900 (depending on the batch) for 10 minutes at + 37 ° C with gentle shaking.
  • the process is repeated twice with washing buffer and twice with phosphate-buffered saline.
  • the substrate (4 chloronaphtol + H 2 ° 2) is produced and briefly preheated at + 37 ° C.
  • Fig.la shows a strong color reaction of the serum dilutions up to 1: 600 when using the
  • Antigen dilutions 1:50 to 1: 200 The 1:50 serum dilution still has a clear reaction with an antigen dilution of 1: 1200. This is also the case with the serum dilutions 1: 100 and 1: 200.
  • the serum dilution 1: 600 was negative with this antigen dilution, but was still positive with an antigen dilution of 1: 400.
  • the black spots shown only partially broken through in the figures are intended to represent weak color reactions.
  • Fig. 1b shows the corresponding reaction with a serum without antibodies against rabies virus.
  • the serum dilutions 1:50 to 1: 200 only show a weak non-specific reaction at the antigen dilutions 1:50 and 1: 100. For this reason, a dilution of 1: 200 was used with this antigen in the subsequent studies.
  • 1c shows the result of a rabies immunoglobulin with a known antibody content.
  • a comparison of this strip with the other strips shows an identical color reaction of the serum dilution of the 1: 100 diluted positive serum. This serum therefore has 1/10 of the antibody content of the immunoglobulin preparation.
  • this example shows that there are quantitative dependencies between antigen and antibody with this method and that a semi-quantitative statement about the antibody and antigen content is possible.
  • FIG. 2 shows the test result that was carried out with the described Blood Drop ELISA.
  • a sample of a test subject of whom the positive rabies virus antibody status was known, was diluted 1:75 in dilution buffer and then a carrier with rabies virus antigen was immersed in this dilution. After the reaction had continued as shown in Example 1, a positive reaction was detected.
  • a drop of capillary blood of approximately 20 ⁇ l from the fingertip of a test subject without rabies virus antibody was dripped directly into a vessel intended for the absorption of the antigen carrier, which contained 1500 / ul dilution buffer. After the course of the reaction described in Example 1, the result was negative.
  • Example 3 Testing Different Carrier Materials for the Blood Drop ELISA.
  • FIG. 3 shows the result of 5 rabies virus antibodies positive and 3 negative sera.
  • the antibody status was determined in the established neutralization test.
  • the figure shows that not only nitrocellulose and chromatography paper are suitable as carriers, but also other papers.
  • the pore size should not be too large so that the antigen does not diffuse too strongly and is therefore only present in a relatively low concentration of antigen per unit area, which leads to a weaker coloration.
  • FIG. 4 shows the results of a screening of 17 risk subjects for rabies immunity using the Blood Drop ELISA. The results were correlated with those of established antibody studies.
  • the blood drop ELISA was used to examine in a drop of blood which virus infections had occurred. These results were examined for validity in the conventional complement fixation reaction and in the ELISA of Behring-Werke, Marburg (Enzygnost). With a carrier and the use of a single drop of blood, determinations with 17 antigens were carried out in about 1/2 hour. The result shown in FIG. 5 showed agreement between the results achieved with the conventional methods.
  • FIG. 6 shows the result of an examination of patient blood samples for antibodies against the human immunodeficiency virus HIV (I), the AIDS pathogen.
  • the HIV antibody status results for these 11 selected individuals are identical in conventional and blood drop ELIZA.
  • the names in the right column mean:
  • Negative no antibodies to HIV present, i.e. not infected
  • HIV antigen is infected cells (H9 cell strain).
  • control antigen is uninfected H9 cells.
  • FIG. 7 shows a section through an incubator according to the invention for the test batch.
  • the incubator shown essentially consists of a double-walled container 10 which has a recess 12 in which one or more carriers can be arranged.
  • the container 10 is closed by a lid 14, so that the test mixture in which the test strip is located can be shaken.
  • the cover 14 is formed from transparent material, so that a color reaction to the test strip can be recognized through the cover.
  • the lid 14 is also designed so that it can be removed and replaced several times.
  • the double-walled container 10 is hollow and can be filled with water 18 via a filler neck 16. This embodiment thus allows a water bath at the desired temperature.
  • FIG. 8 shows a sectional view through another incubator.
  • the container 20 shown in this Fig. 8 consists of a good heat-conducting material.
  • a test mixture 23, which surrounds a test strip 25, is again contained in the container.
  • the container has on the outside of its upper region a locking projection 27, under which a locking projection 29, which is formed on the downwardly projecting skirt of the lid 24, projects inwards .
  • the container 20 can be closed by the lid 24 so that the latter snaps onto the container 20.
  • Such an incubator can easily be shaken by hand without test liquid falling out, and at the same time the lid can be easily removed so that the test mixture can be poured off or the strip can be removed.
  • This embodiment of the invention has the advantage that it manages without additional heat sources.
  • Such an incubator can first be put in your pocket until it reaches body temperature. The examination can then be carried out.
  • the carrier is inserted and also when the test mixture is poured in or poured out, the incubator remains at temperature because the test person is holding it in the hand.
  • the carrier is inserted and also when the test mixture is poured in or poured out, the incubator remains at temperature because the test person is holding it in the hand.
  • This incubator also does not cool down during the reaction time: either, the user holds this incubator in hand, e.g. to shake him, or he just puts it back in his pocket. In
  • FIG. 9 shows another incubator 30, which essentially consists of heat-conducting material 35, in which heating wires 36 are embedded. This embodiment of the invention enables the test to be carried out at constant temperature simply by connecting it to a battery.
  • FIG. 10 shows a modification of the incubator of FIG. 9.
  • a disposable insert 41 is shown, which lines the recess. This disposable or disposable insert receives the test liquid and the test strip 46. After a test has been carried out, the disposable insert 41 and its contents are removed. The incubator can then be used for the next examination.
  • This incubator 40 also has heating walls or heating wires 47. Furthermore, a thermostat 48 is provided which ensures that the incubator 40 is always kept at a constant temperature. The entire incubator is arranged on a shaker 49.
  • Fig. 11 shows an illustration for carrying out the AIDS test for lay people.
  • the figure shows a strip or carrier on which 'HIV + cells', marked by 1, and 'HIV cells', marked by 2, are applied.
  • 1 thus denotes virus antigen
  • 2 denotes control antigen.
  • FIG. 12 shows an interpretation aid for evaluating the result.
  • the result of the investigation is only positive if 1 is colored much more strongly than 2.
  • the Coloring of 1 must be stronger than in the example of Fig. 12 c.
  • the results of the findings are shown in each of the illustrations in FIGS. 12 a, b, c and d, in cases a and b the findings are positive, represented by a "+", in FIG. 12 c the findings are questionable, represented by a " ? ", and in FIG. 12d the result is negative, represented by a" - ".
  • the finding is questionable, in 12f it is positive and in 12g and 12h the finding is negative.
  • the identification of a finding by "+” means that antibodies against HIV I are present.
  • the finding "-" means that antibodies against HIV I are not present.
  • the finding "?” states that the result of the finding is questionable. There are probably no antibodies. In this case, repeating the examination or consulting a doctor is recommended.
  • FIG. 13 shows another incubator which can also be used by the layperson in a very simple manner.
  • This incubator 50 has a rigid inner container 52 which is surrounded by an outer, flexible plastic bag 54 which is filled with heat-storing material 56.
  • the inner rigid container 52 is thus closed on the left side by a plug-like cover 58.
  • This embodiment of the invention has the advantage that the incubator can be filled with, for example, granular or pasty material. Such a body will adapt to the hand when it is picked up. If the body is pressed a little, the convex shapes of the fingers will also press into the surface of this incubator, so that a conceivably large contact surface between the outer circumference of the incubator and the hand is achieved. This advantageous result is achieved regardless of whether the person carrying out this examination has a large or a small hand. This ensures optimal heat transfer between the hand and the incubator. In this way, when the material 56 is thermally conductive, the inner container and the test strip arranged in the container and the test mixture are always kept at the same temperature.
  • FIG. 13 which shows an 'vertical section
  • a holder 58 on which one or more test strips are attached.
  • FIG. 14 shows a top view of the incubator of FIG. 13, partly in section. The same parts again have the same reference numerals.
  • a test strip 60 as shown in FIGS. 11 and 12, is arranged on the holder 58, which is angled and has a handle-like projection 59 at its left end.
  • FIG. 15 shows another embodiment of an incubator which is for sale in an airtight manner final, welded sleeve 79 is packaged.
  • a recess 72 is again provided, which is closed by a cover 74.
  • the incubator 70 has, outside the recess 72, recesses 75 which can be designed as a groove.
  • a projection protruding downward from the cover 74 engages in this locking recess 75.
  • the cover 74 can be snapped onto the incubator 70; the cover can be easily removed from the incubator 70 at any time via its lateral projections 73 which protrude beyond the recesses 75.
  • FIG. 15 it can be seen in the sectional illustration in FIG. 15 that two further, approximately cylindrical recesses 64 are provided in the incubator 70. Containers 86 and 88 are provided in these recesses, which can contain the test mixtures required for the examination. 15 only two such further recesses 82 and 84 are shown. However, as can be seen in FIG. 16, four or more such recesses can be provided.
  • the liquid containers 86 and 88 which are contained in the recesses 82 or 74, can consist of glass, which has a predetermined breaking point, for example, at its pointed end. In this way, even the layperson can introduce the test solutions or test mixtures into the recess 72 in a very simple manner.
  • these containers 86 and 88 are made of a flexible material that has a predetermined breaking point, for example, at its front tip. This development of the invention has the advantage that the containers 86 and 88 serve multiple uses. First of all, they store the test mixtures. During the examination, they enable the test mixture contained to be brought into the recess 72 with ease and without any specialist knowledge.
  • FIGS. 15 and 16 thus represents a marketable unit which can be kept indefinitely because it is sealed airtight by the sleeve 79 and contains all the components which are necessary for carrying out an examination.
  • FIG. 15 also shows a holder 80 which can hold the test strips.
  • This holder is shown in Figures 17 and 18 and is described in more detail with reference to these figures.
  • FIG. 80 A number of test strips are arranged on the container.
  • a handle 90 On the right edge of holder 80 there is a handle 90 which has a predetermined breaking point at 92. If this holder is inserted into the recess of an incubator, for example in the recess 72, the handle can be easily broken off so that the recess can be closed off by the cover, for example 74. In this way, it is possible in a very simple manner to insert the test strips into the recess.
  • test strips are conveniently clamped in the holder 80, e.g. how a slide is clamped in a frame. In this way it is possible to carry out the examinations without touching the nitrocellulose, which essentially consists of the test strip, with the fingers. This is particularly important if the investigations are to be carried out by the layperson.
  • test strips or supports made of nitro-cellulose have been offered without such a frame. This is still possible because until now such carrier strips have only been used in laboratories in which the necessary tools such as tweezers and plastic pipettes and the like are available. According to the invention, these tools no longer have to be present, since the holder can be gripped by the handle 90 and the test strips are clamped in the holder.
  • the nitrocellulose does not have to be touched with the fingers to apply the test mixtures either, since, as in the exemplary embodiment in FIG. 15, they can be supplied in plastic containers 86, 86.
  • the holder 80 can be packed in a tub, cf. Fig. 18, which can be inserted into the recess of an incubator. In this way it is ensured that the test strip does not have to be touched with the fingers.
  • the incubator there can be used as often as desired.
  • the incubator shown there can be used as a finished laboratory device with all
  • ingredients and test mixes are sold. Furthermore, it is possible, as additional kits, to have the containers 86 or 88, in the form of vials or small plastic containers, and the tub 94, as shown in FIG. 18, together with the carrier and test strips located thereon, as replacement components to put on the market.
  • the trough 94 in FIG. 18 can of course also be packed in an airtight manner, so that the test strips cannot be influenced subsequently by touching them with fingers or by dirt.
  • an identification aid 96 is also attached to the left edge. ' This can consist of a printed strip, which is the key to evaluating the test result. Such decryption values are given by way of example by handwriting under the holder 80 in FIG. 17.

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Abstract

Un procédé permet de dépister des protéines à propriétés spécifiques de type anticorps ou antigènes dans des fluides corporels ou d'autres matières organiques. On utilise une protéine fixée sur un support qui entre en réaction spécifique avec la protéine que l'on veut dépister dans la solution, rendant cette dernière insoluble, et une deuxième protéine se trouvant dans la solution, pourvue d'un marqueur, qui entre en réaction avec le composant insoluble et devient elle aussi insoluble. On peut ainsi examiner directement du sang, sans centrifugation préalable, afin d'y dépister simultanément une ou plusieurs protéines.
PCT/EP1988/000693 1987-07-31 1988-07-31 Procede et dispositif d'examen de sang complet, d'autres fluides corporels et d'autres fluides biologiques WO1989001156A1 (fr)

Applications Claiming Priority (2)

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DEP3725504.5 1987-07-31
DE19873725504 DE3725504A1 (de) 1987-07-31 1987-07-31 Verfahren und vorrichtung zum nachweis von antikoerpern und antigenen

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WO1989001156A1 true WO1989001156A1 (fr) 1989-02-09

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AU (1) AU2088188A (fr)
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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
LT3644B (en) 1990-03-19 1996-01-25 Ina Acquisition Corp Envelope pipe and method for covering of pipeline or kanal

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE3842700A1 (de) * 1988-12-19 1990-06-21 Boehringer Mannheim Gmbh Verfahren zur proteinimmobilisierung an einer festphase, so hergestellte protein tragende festphase sowie deren verwendung
DE4243375C2 (de) * 1992-12-21 1998-04-23 Brahms Diagnostica Gmbh Bestimmungsverfahren zur Bestimmung von Antikörpern in biologischen Flüssigkeiten

Citations (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3579306A (en) * 1969-01-22 1971-05-18 Organon Diagnostic test device
GB1414479A (en) * 1971-12-21 1975-11-19 Abbott Lab Test apparatus for direct radioimmunoassay for antigens and their antibodies
US4147752A (en) * 1977-01-14 1979-04-03 Kommandiittihytio Finnpipette Osmo A. Souvaniemi Form piece for apparatuses used for immunoassays and enzyme reactions
GB1565402A (en) * 1975-11-03 1980-04-23 Int Diagnostic Tech Diagnostic reagent holder
EP0113075A2 (fr) * 1982-12-06 1984-07-11 Fielder Gillespie Davis Limited Système de réaction d'essai immunologique transportable, procédé pour le diagnostic par essai immunologique et sonde ou support utilisé dans ce procédé
EP0160467A2 (fr) * 1984-04-20 1985-11-06 Syntex (U.S.A.) Inc. Essai immuno-enzymatique, support absorbant pour sa mise en oeuvre et dispositif d'essai
EP0194789A2 (fr) * 1985-02-28 1986-09-17 Becton Dickinson and Company Dispositif et procédé pour le dosage multiple simultané
EP0201339A2 (fr) * 1985-05-09 1986-11-12 Ultra Diagnostic Corporation Méthode d'essai immunologique, dispositif et trousse de réactifs
EP0204579A2 (fr) * 1985-06-05 1986-12-10 Synbiotics Corporation Dispositif d'incubation des essais immunologiques
EP0269092A2 (fr) * 1986-11-26 1988-06-01 Roche Diagnostics GmbH Procédé pour la détermination d'une substance de liaison spécifique

Family Cites Families (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4148869A (en) * 1975-03-06 1979-04-10 Baxter Travenol Laboratories, Inc. Immunological reagent and method of using same
US4452903A (en) * 1981-02-17 1984-06-05 Lee Jin P Assay method and reagent kit means for lipid-containing body fluid
EP0061541A1 (fr) * 1981-03-27 1982-10-06 Biospecia Limited Analyse immunologique et agent biochimique pour cette analyse
GB2121962B (en) * 1982-06-08 1985-10-02 Serono Diagnostics Ltd Manual immunoassays and apparatus therefor
DE3327642A1 (de) * 1983-07-30 1985-02-14 Boehringer Mannheim Gmbh, 6800 Mannheim Verfahren zur bestimmung eines partners einer immunreaktion sowie reagens zur durchfuehrung dieses verfahrens
US4594327A (en) * 1983-11-02 1986-06-10 Syntex (U.S.A.) Inc. Assay method for whole blood samples

Patent Citations (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3579306A (en) * 1969-01-22 1971-05-18 Organon Diagnostic test device
GB1414479A (en) * 1971-12-21 1975-11-19 Abbott Lab Test apparatus for direct radioimmunoassay for antigens and their antibodies
GB1565402A (en) * 1975-11-03 1980-04-23 Int Diagnostic Tech Diagnostic reagent holder
US4147752A (en) * 1977-01-14 1979-04-03 Kommandiittihytio Finnpipette Osmo A. Souvaniemi Form piece for apparatuses used for immunoassays and enzyme reactions
EP0113075A2 (fr) * 1982-12-06 1984-07-11 Fielder Gillespie Davis Limited Système de réaction d'essai immunologique transportable, procédé pour le diagnostic par essai immunologique et sonde ou support utilisé dans ce procédé
EP0160467A2 (fr) * 1984-04-20 1985-11-06 Syntex (U.S.A.) Inc. Essai immuno-enzymatique, support absorbant pour sa mise en oeuvre et dispositif d'essai
EP0194789A2 (fr) * 1985-02-28 1986-09-17 Becton Dickinson and Company Dispositif et procédé pour le dosage multiple simultané
EP0201339A2 (fr) * 1985-05-09 1986-11-12 Ultra Diagnostic Corporation Méthode d'essai immunologique, dispositif et trousse de réactifs
EP0204579A2 (fr) * 1985-06-05 1986-12-10 Synbiotics Corporation Dispositif d'incubation des essais immunologiques
EP0269092A2 (fr) * 1986-11-26 1988-06-01 Roche Diagnostics GmbH Procédé pour la détermination d'une substance de liaison spécifique

Non-Patent Citations (5)

* Cited by examiner, † Cited by third party
Title
CLINICAL CHEMISTRY, Vol. 24, No. 3, 1978, pages 455-59, J.C. FRUCHART et al., "Enzyme Immunoassay for Human Apolipoprotein B, the Major Protein Moiety in Low-Density- and Very-Low-Density Lipoproteins". *
CLINICAL CHEMISTRY, Vol. 30, No. 11, 1984, pages 1753-57, P. KASPAR et al., "New Photometric Assay for Chymotrypsin in Stool". *
DIALOG, Access. Number 085248, Analytical Abstracts, AA Access. Number: 49-02-D-00005, Immunoassay Tech. Nol., Volume 2, Issue 189-213, Publication Date 860000. *
JOURNAL OF IMMUNOLOGICAL METHODS, 52, (1982), 395-408, R.G.E. PALFREE, "An Enzyme-Linked Immunosorbent Assay (Elisa) for Detergent Solubilized Ia Glycoproteins Using Nitrocellulose Membrane Discs". *
MEDLINES, NLM, Access. Number 86079998, CLIN. CHEM., January 1986; 32(1 Pt 1): 16-21. *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
LT3644B (en) 1990-03-19 1996-01-25 Ina Acquisition Corp Envelope pipe and method for covering of pipeline or kanal

Also Published As

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EP0327629A1 (fr) 1989-08-16
DE3725504C2 (fr) 1989-10-05
AU2088188A (en) 1989-03-01
DE3725504A1 (de) 1989-02-09

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