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
  • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
  • G01N33/6887—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids from muscle, cartilage or connective tissue
• • G—PHYSICS
  • G01—MEASURING; TESTING
  • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
  • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
  • G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
  • G01N2333/46—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans from vertebrates
  • G01N2333/47—Assays involving proteins of known structure or function as defined in the subgroups
  • G01N2333/4701—Details
  • G01N2333/4712—Muscle proteins, e.g. myosin, actin, protein
• • G—PHYSICS
  • G01—MEASURING; TESTING
  • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
  • G01N2800/00—Detection or diagnosis of diseases
  • G01N2800/52—Predicting or monitoring the response to treatment, e.g. for selection of therapy based on assay results in personalised medicine; Prognosis

Definitions

• troponin is a myofib llar protein complex, made up of three proteins, troponins I, T and C. This protein complex helps to regulate muscle contraction by the Ca 2+ ion, interacting with myosin and actin. More precisely, it is known that when a nerve impulse arrives at the level of the motor plaque of a muscle, there is generation of an action potential which is transmitted to the sarcoplasmic reticulum. Ca 2+ is then released into the cytosol and binds to troponin C, which leads to a strengthening of the interaction between troponin I and troponin C and, consequently, a change in conformation of the troponin I, T complex. , C. There is then release of the actin-myosin interaction sites, which allows the muscle to contract.
• heparin in addition, before angioplasty or after a myocardial infarction or during certain treatments for cardiovascular conditions, patients are administered significant amounts of heparin and this treatment is often extended beyond 24 hours. Depending on the dose administered, the concentration of heparin in the plasma one hour after administration can vary between 0.1 to 2 IU / ml. The presence of heparin in the blood samples due to the treatment of the patient constitutes an important problem for the dosage of troponins. - 3 -
• the present invention relates more specifically to an immunological assay method for troponin I (cardiac or skeletal), T or C, and troponins I, T or C associated together in the form of dimers (IT, IC or TC) or in the form trimer (ITC) in a biological sample containing heparin, characterized in that the immunoassay is carried out in the presence of hexadimethrin bromide (polybrene).
• hexadimethrin bromide polybrene
• the amount of polybrene used during the immunoassay of troponin I (cardiac or skeletal), T or C, and troponins I, T or C associated with one another in the form of dimers (IT, IC or TC) or under Trimer form (ITC) may vary depending on the immunoassay procedure used.
• the ratio [molar concentration of the polybrene used / molar concentration of heparin present in the sample to be analyzed] can be from 1 to 1000. Depending on the dosage, it can more specifically be - 4 -
• the present invention can be implemented with any immunological method allowing the evaluation of troponin I (cardiac or skeletal), T or C, and troponins I, T or C associated with one another in the form of dimers (IT, IC or TC) or in the form of trimer (ITC).
• troponin I cardiac or skeletal
• T or C troponins I, T or C associated with one another in the form of dimers (IT, IC or TC) or in the form of trimer (ITC).
• troponin I cardiomyalpha-1 (cardiac or skeletal), T or C, and troponins I, T or C associated together in the form of dimers (IT, IC or TC) or in the form of trimer (ITC), to avoid interference due to heparin is part of the present invention
• heparin lithium heparinase I
• heparinase I heparinase I from Flavobacterium heparinum Sigma ref. H 2519
• L-histidine Sigma ref. H 8000
• protamine sulfate protamine sulfate grade X from salmon Sigma ref. P 4020
• antithrombin III antithrombin III from huma ⁇ plasma Sigma ref. A 7388
• cation exchange resins QSFF (Q Sepharose Fast Flow, Pharmacia Biotech ref. 17051001) and DEAE (DEAE Sepharose Fast Flow, Pharmacia Biotech ref. 17070901).
• the total analysis time is 15 minutes.
• the Troponine I Pasteur kit was used (Sanofi Diagnostics Pasteur, Marnes-la Coquette, France). The assay is carried out as follows:
• washing is carried out by adding 1 ml of 0.1 M phosphate buffer, pH 6.8 containing 0.1% Tween® 20 and 0.3% Kathon, keeping the temperature of the washing solution at most 8 ° C. This step is repeated 4 times.
• the enzymatic development is carried out using 600 ⁇ l of a mixture consisting of 1 part of tetramethylbenzydine and 100 parts of citrate buffer containing 4% DMSO and 0, 03% hydrogen peroxide.
• Tnl positive serum + diluent 1 160 1,240 Tnl positive serum + heparin 40IU / ml 0.480 1.170
• the troponin I assay is performed with the Troponine I Pasteur kit (Sanofi Diagnostics Pasteur, Marnes-la Coquette, France).
• the automated system used for the enzyme immunoassay is the Access® immuoassay system, a system marketed by Beckman.
• the immunoassay method is that described in Example I.

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• General Health & Medical Sciences (AREA)
• General Physics & Mathematics (AREA)
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• Investigating Or Analysing Biological Materials (AREA)

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• 1998
  • 1998-02-05 FR FR9801367A patent/FR2774473B1/fr not_active Expired - Fee Related
• 1999
  • 1999-02-01 JP JP2000530804A patent/JP2002502979A/ja active Pending
  • 1999-02-01 WO PCT/FR1999/000198 patent/WO1999040442A1/fr not_active Application Discontinuation
  • 1999-02-01 EP EP99901673A patent/EP1051623A1/de not_active Withdrawn

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