Long-optical-path enzyme-linked immunoassay for testing thyroid stimulating hormone, and kit
A thyroid-stimulating hormone and long optical path technology, which is applied in the direction of analytical materials, biological testing, measuring devices, etc., can solve problems such as insufficient sensitivity, achieve the effects of expanding the measurement range, increasing the amount of coated antibodies, and improving the performance of immunoassays
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preparation example Construction
[0097] The preparation method of above-mentioned kit comprises the following steps:
[0098] (1) Preparation of antibody-coated tubes
[0099] Dilute the TSH antibody to 0.6 mg / L with 0.03 M, pH 7.2 phosphate buffer, add 200 μL to each polystyrene tube, and keep overnight at 4 °C; discard the reaction solution, and add 1% bovine serum white to each tube Protein 0.03M pH 7.2 phosphate buffer 300ml, block at 37°C for 2 hours, discard the blocking solution, freeze or dry at room temperature, store at 4°C for use;
[0100] (2) Preparation of TSH series standard products
[0101] Dilute the pure TSH product with calf serum containing 0.2% thimerosal and inactivate at 56°C for 1.0 hour to a series of standard products with concentrations of 0, 0.05, 0.3, 4.0, 10, and 20 mIU / L, and pack in 1.0 ml per bottle , stored at -20°C for use;
[0102] (3) Preparation of enzyme markers
[0103] Weigh 5mg of horseradish peroxidase, dissolve in 1ml of distilled water, fully dissolve; add 1ml...
Embodiment 1
[0130] The TSH long-path ELISA method provided by the present invention is compared with the conventional TSH ELISA method: the conventional TSH ELISA method is established by using the same antibody as in the present invention, and the sensitivity is 0.06mIU / L after methodological optimization. , the linear range is 0.3~20mIU / L; the sensitivity of the analytical method of the present invention is 0.015mIU / L, and the linear range is 0.05~20mIU / L; the sensitivity of the analytical method of the present invention is improved by 4 times compared with the conventional ELISA method, and has obvious Sensitivity, linear range advantage (see Table 2 below). The linear range in the table refers to the range between the standard point where the first concentration is not zero and the standard point of the highest concentration; the signal-to-noise ratio refers to the ratio between the standard point where the first concentration is not zero and the zero standard point; Linear correlatio...
Embodiment 2
[0134] According to the TSH analysis method and operation steps of the present invention, a 0.5m fused silica tube was used as a flow sample cell to carry out an immunoassay experiment to obtain a better standard curve, sensitivity and signal-to-noise ratio (see Table 3).
[0135] Table 3 Experimental results of 0.5m fused silica tube flow sample cell
[0136]
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