• 专利附录
  • 专利说明
  • 权利要求
  • 类似技术
  • 同族专利
  • 引用文献
  • 法律状态
  • 优先权
    • 专利摘要:
    A method for the detection of hemoglobin or decomposition products of hemoglobin in feces. Human hemoglobin is isolated from the sample by means of an immunological reaction, i.e. a reaction between an antibody and an antigen, by using an antibody specific for human hemoglobin and attached to the solid phase. The human hemoglobin bound to the antibody is established hereupon.
    公开号:SU1496643A3
    申请号:SU813235497
    申请日:1981-01-16
    公开日:1989-07-23
    发明作者:Суованиеми Осмо；Адлеркреуц Херман；Суни Юкка；Партанен Пол
    申请人:Коммандииттиихтие Финнпипетте Осмо А.Суованиеми (Фирма)；
    IPC主号:
    专利说明:

    The invention relates to medicine, more specifically to biochemical methods for the detection of hidden blood in the diagnosis of various clinically difficult to diagnose intestinal diseases, such as neoplasm, etc. my guts, adenomas, ulcers and diverticula.
    The purpose of the invention is to increase the specificity and sensitivity of the determination.
    The method is based on a two-phase occult blood test, combining a highly sensitive assay using gua cum resin with a highly specific and sensitive immunochemical assay, which is used to determine only human hemoglobin. This combination of tests eliminates false positive results of the analysis, as well as the need to control the food regime.
    The equipment required for the proposed method includes a bag for analyzing feces into which the sample is placed, and a second bag having a paper filter. The absorbent (paper filter) is attached to the bag envelope so that it is between the impregnated gum resin, the pouch paper and the plastic cap.
    When the hemoglobin is extracted from the absorbent, a solution containing hemoglobin (HB) and other soluble molecules from the feces is obtained, this solution is pipetted into a cuvette from a set of polystyrene cuvettes

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    creHKVi and at the bottom of which a certain amount of human hemoglobin antibody is applied. Here, the sample hemoglobin (antigen) binds to the solid phase antihemoglobin (IgG), after an incubation period of the same time as the immunological reaction, the cuvette is washed with water to remove all unrelated substances.
    The solid phase may be a polymer other than polystyrene, and may have a different shape than the cuvette or vessel, for example, a ball, a ring, a disk.
    I After the cuvette is washed, a certain amount of human hemoglobin antibody IgG is placed in it, while the IgG has some enzyme as an indicator. The binding of an enzyme-labeled IgG to an antibody in the solid phase depends on the amount of free antibodies in the solid phase remaining after the sample is processed. Thus, using an enzyme, one can check whether a reaction between an antibody (human anti-Hb) and an antigen (human Hb) was done by adding a substrate that is specific for this enzyme, while the enzyme removes a colored visible or photometrically recorded compound from the substrate. The amount of enzyme can be determined either by measuring the amount combined with the solid phase or the amount remaining in the solution free.
    . The FECA-EIA assay detects only intact human hemoglobin, and the gum resin assay also detects hemoglobin breakdown products. In addition, errors caused by the presence of animal hemoglobins inherent to the known methods are eliminated.
    Example. From the laboratory side of the package case cover, an absorbent containing the sample is taken and placed in a cuvette of the FP-9 analyzer. The FP-9 polystyrene cuvette was pre-sensitized with an immuno-globulin fraction of anti-hemoglobin. 200 µl of phosphate-containing buffer solution (0.1 M polybutadiene, 4) was added to the cuvette. Incubation, for example, 2 hours at C. The cuvette is washed with 400 µl of distilled water. The cuvette is added
    200 µl of the human anti-hemoglobin immunoglobulin fraction (polybutadiene styrene as solvent) with a suitable enzyme, for example, alkaline phosphatase (conjugate addition). Incubation, for example, for 2 hours at. Washing is similar. A substrate specific for the enzyme that is used in the conjugate (para-nitrophenylphosphate) is added. After 30 min incubation at 200, 200 µl of 0.1 M NaOH is added to stop the reaction. The result is evaluated photometrically.
    The recommended limit for stopping the reaction is the average value of absorption (minus the control value), normal non-flowing samples + 3 standard deviations, or in the absence of such data - 9,200 units.
    The smallest amounts detected by this analysis are 0.05-0.1 µg cyanmethemoglobin (ml 1-5 µl hemolyzed blood per 1 g feces), 140-700 µg hemoglobin (g feces) and from 2.0 to 5.0 µl whole blood (g feces, (280-700 µg. Hemoglobin (g feces)
    权利要求:
    Claims (1)
    [1]
    Invention Formula
    The method of determining human hemoglobin in faeces by immunochemical analysis, characterized in that, in order to increase the specificity and sensitivity of the method, 1 g of the sample is adsorbed through paper soaked in glue resin on filter paper, the filter paper is placed in the measuring cell , pretreated for 15 hours with the immunoglobulin fraction of human antihemoglobin, 200 μl of phosphate buffer solution pH 7.4, containing 0.1 M polybutadiene styrene, are added, incubated 2 at 37 ° C, wash the well with 400 µl of distilled water, add 200 µl. the solution of the immunoglobulin fraction of human antihemoglobin, combined with alkali phosphatase and dissolved in the phosphate buffer solution, is incubated for 2 hours at 37 ° C, washed again, 200 micron para-nitrophenylphosphate is added and after incubation 14966436
    For 30 minutes at 405, they determine the general level of the reaction by using 200 µl of the human hemoglobin, M NaOH solution and by absorbing the bin in the sample.
    类似技术:
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    同族专利:
    公开号 | 公开日
    DE3172465D1|1985-11-07|
    JPH0121907B2|1989-04-24|
    FI800140A|1981-07-18|
    EP0032782B1|1985-10-02|
    EP0032782A3|1982-01-27|
    FI61965C|1982-10-11|
    CS228136B2|1984-05-14|
    EP0032782A2|1981-07-29|
    JPS56106154A|1981-08-24|
    US4427769A|1984-01-24|
    DD157025A5|1982-10-06|
    AT15945T|1985-10-15|
    FI61965B|1982-06-30|
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    法律状态:
    优先权:
    申请号 | 申请日 | 专利标题
    FI800140A|FI61965C|1980-01-17|1980-01-17|DETECTING OF HEMOGLOBIN AND DETECTION|
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