Cardiac troponin I assay reagent and method
A technique for cardiac troponin and reagents, applied in the field of cardiac troponin I assay reagents
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Embodiment 1
[0091] [Example 1] Preparation of Cardiac Troponin I Determination Reagent, Measurement Method and Preparation of Sample to be Tested
[0092] As cardiac troponin I measurement reagents, the following reagents A and B with different antibody pairings were prepared.
[0093]
[0094] Antibody-binding particle solution (immobilized antibody solution): Prepare antibody-binding magnetic particles containing 0.025% (w / v), 50 mM 2-morpholineethanesulfonic acid (MES), 1.0% (w / v) Serum albumin (BSA) and 50 mM NaCl antibody-bound particle solution (pH 6.8) that binds to cardiac muscle as an epitope on carboxylated magnetic particles (manufactured by Fuji REBIO Co., Ltd.) The amino acid sequence of troponin I (for example, refer to GenBank: CAA62301.1, the same applies hereinafter) was obtained by recognizing mouse monoclonal antibody at positions 41 to 49.
[0095] Labeled antibody solution: Prepare labeled antibody containing 0.5 μg / mL, 50 mM MES, 2.5% (w / v) BSA, 100 mM NaCl, 0.3 m...
Embodiment 2
[0108] [Example 2] The impact of dextran sodium sulfate on serum plasma correlation in cardiac troponin I assay
[0109] Dextran sodium sulfate (dextran sodium sulfate 5000, manufactured by Wako Pure Chemical Industries, Ltd.) was added to the antibody-conjugated particle solution of reagent A at 0 mg / mL, 0.77 mg / mL, and 1.55 mg / mL , 2.32mg / mL (concentrations in the first reaction liquid are respectively 0mg / mL, 0.462mg / mL, 0.930mg / mL, or 1.392mg / mL), measure the sample to be tested. The results are shown in Table 1 and Figure 1 ~ Figure 3 middle. Each measured value is the emission intensity (count value) at a wavelength of 477 nm. For each sample, the percentage of each plasma count value to the serum count value was calculated, and then their average value was calculated to obtain a paired serum count value (mean). The paired serum count values (mean) in Example 3 and Example 4 are values obtained in the same manner.
[0110] [Table 1]
[0111] Table 1. Determinat...
Embodiment 3
[0116] [Example 3] Influence of the type of antibody epitope on serum plasma correlation in the measurement of cardiac troponin I in the presence of dextran sodium sulfate
[0117] Dextran sodium sulfate (dextran sodium sulfate 5000, manufactured by Wako Pure Chemical Industries, Ltd.) was added to the antibody-conjugated particle solution of reagent B at 0 mg / mL, 0.5 mg / mL, 1.0 mg / mL, Or 2.0mg / mL (concentration in the 1st reaction liquid is respectively 0mg / mL, 0.3mg / mL, 0.6mg / mL, 1.2mg / mL), other conditions are identical with embodiment 2, and test sample is measured. The results are shown in Table 2 and Figure 4 ~ Figure 7 middle. Each measured value is the emission intensity (count value) at a wavelength of 477 nm.
[0118] [Table 2]
[0119] Table 2. Determination of Cardiac Troponin I Using Reagent B
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[0121]
[0122] As a result, reagent B was used, which recognized the cardiac troponin I antigen using a different antibody pair (immobilized antibody ...
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