Beta 2-microglobulin detection kit
A detection kit and microglobulin technology, which is applied in the field of medical immunology, can solve the problems of false positive test results, antibody shedding, false elevation, etc., and achieve the effects of high accuracy, reduced dosage, and low production cost
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Embodiment 1
[0030] The preparation of embodiment 1β2-microglobulin detection kit
[0031] The kit of the present invention relates to the main raw materials of reagents as follows:
[0032] 1. Antibody to β2-microglobulin: the titer was determined by indirect ELISA method to be 1:100000.
[0033] 2. Latex: The present invention only exemplarily uses polystyrene latex particles with a diameter of 200-300 nm and carboxyl groups to carry out experiments.
[0034] The preparation of the main reagent of this embodiment is as follows:
[0035] Reagent R1: 0.2M ammonium chloride buffer containing 1.0% PEG6000 (polyethylene glycol 6000), 0.09% sodium azide, 0.2% BSA, 0.8% NaCl, 0.3% Tween 20. The reagent is a colorless transparent solution.
[0036] Reagent R2: 0.1M Tris buffer containing 0.2% β2-microglobulin antibody sensitized polystyrene latex particles, 1% BSA, 0.2% NaN 3 . The reagent is a milky white solution. Specific steps are as follows:
[0037] 1. Take 1ml (100mg / ml) latex, was...
Embodiment 2
[0047] Determination of embodiment 2β2-microglobulin
[0048] Detection tool: Hitachi 7060 automatic analyzer.
[0049] Analysis method: two-point endpoint method; main wavelength: 546nm, secondary wavelength: 700nm: sample volume: 3.0ul; reagent R1: 240ul; reagent R2: 60ul; calibration method: spline function Spline; reaction direction: rising; measurement temperature: 37°C; after mixing the sample and reagent R1, read the absorbance A at 30 seconds 1 ; Add reagent R2 at 5 minutes, read absorbance A at 5 minutes 2 . The absorbance of the reaction was calculated as A 2 with A 1 calibration difference.
[0050] Calculation method: multi-point calibration, using spline function as the calculation mode, and making a dose / response curve according to the absorbance and reference serum value, and the sample content can be calculated on the dose / response curve according to its absorbance value.
[0051] Reference range: 1.0~3.0mg / L
Embodiment 3
[0052] Example 3 Testing of various performance indicators of the β2-microglobulin detection kit
[0053] 1. Sensitivity test
[0054] Measure the sample of 6 kinds of different β2-microglobulin contents, each sample measures 10 times, by calculating mean value and SD value (standard deviation value), as seen from Table 1, the sensitivity of detection kit of the present invention is 0.08mg / L.
[0055] Table 1
[0056] β2-MG(mg / L)
[0057] 2. High value linear determination
[0058] Measure 10 samples of different β2-microglobulin content, each sample is measured 2 times, as can be seen from Table 2, the highest detection range of the detection kit of the present invention can reach 18mg / L, judge basis r 2 ≥0.990.
[0059] Table 2
[0060] Theoretical value of β2-MG (mg / L)
[0061] 10
[0062] 3. Precision test
[0063] Two human serum samples with different β2-microglobulin contents were used to determine the intra-assay precision of the ...
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