Latex enhanced immunoturbidimetry NGAL detection kit
A detection kit and immunoturbidimetric technology, which is applied in the field of latex-enhanced immunoturbidimetric NGAL detection kits, can solve the problems of high reagent background detection linear range, poor reagent stability, and poor anti-interference ability, and achieve The effect of maintaining uniformity, maintaining stability, and wide linear range
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Embodiment 1
[0034] This embodiment relates to a NGAL detection kit and its preparation:
[0035] 1. Reagent R1:
[0036] pH7.0~7.6, 50~400mM buffer
MES buffer
pH7.4, 150mmol / L
Surfactant
Tween-20
50ml / L
Chelating agent
50mmol / L
Accelerator
Dextran 6000
200mmol / L
preservatives
NaN 3
1‰
[0037] Various ingredients can be added sequentially at room temperature, sealed and stored for later use. Alternatively, they are packaged individually and prepared immediately prior to testing.
[0038] 2. Reagent R2 is prepared as follows:
[0039] Step 1: The hydroxyl latex with a particle size of 80 nm was mixed with the hydroxylated latex of 160 nm, and activated with p-toluenesulfonyl cyanogen bromide chloride (50 mg / ml) in MOPSO (40 mM) solution at pH 6.0.
[0040] Step 2: Wash to remove the p-toluenesulfonyl chloride cyanogen bromide in the above solution, add pH 6.0 MOPSO (40mM) buffer, add ...
Embodiment 2
[0048] The NGAL detection kit described in this example is applicable to various types of automatic biochemical analyzers. Taking Hitachi 7170 automatic biochemical analyzer as an example, its operation is shown in Table 1. Analysis method: Two-point endpoint method, that is, the dosage of reagent R1 and reagent R2 is 150ul and 50ul respectively, and the sample volume is 3ul; 150ul reagent R1 is added to 3ul sample, and 50ulR2 is added after 3min at 37°C, that is, the point is read, and the reaction is read after 7min Another point; the main wavelength of the detection wavelength is 570nm.
[0049] Adopt this reagent and above-mentioned assay method, adopt the curve of the NGAL standard substance (self-made) of 6 kinds of different contents that Hitachi 7170 biochemical analyzer records (such as figure 1 Shown), each point represents a content of the reference standard, wherein the X-axis represents the NGAL content (ng / ml); Y-axis represents the absorbance.
[0050] Table 1 ...
Embodiment 3
[0052] Embodiment 3: Correlation test of detection reagent
[0053] Use the inventive reagent of this method (the specific formula is the same as embodiment 1) and the NGAL latex-enhanced immune turbidimetric reagent of a well-known company as a contrast reagent, adopt automatic 7170 full-automatic biochemical analyzer to test 50 parts of human urine (normal and abnormal specimens) by The respective parameters were measured at the same time, and the correlation analysis was carried out on the measured values. Measure according to the parameters in the above-mentioned NGAL determination method, and the determination results are shown in figure 2 , X, Y axis are measured value (the content ng / mL of NGAL), by figure 2 The results show that the correlation between the two reagents is R 2 =0.9914, the regression equation is y=0.9855x+0.9299. The results show that this reagent has a good correlation with a well-known reagent in the determination of patient urine, and has good...
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