Rheumatoid factor detection kit and detection method thereof
A detection kit and rheumatoid factor technology, applied in the biological field, can solve the problems of low and non-typing detection sensitivity, etc., and achieve the effect of strong anti-interference ability, high sensitivity, and wide linear range
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preparation example Construction
[0047] 1. The preparation method of the kit is as follows:
[0048] 1) Reagent R1—magnetic particles coated with rabbit IgG, the specific process is as follows:
[0049] Antibody biotinylation: Take an appropriate amount of rabbit IgG, add an appropriate amount of biotin ester, mix and react on a mixer at room temperature for 0.5 to 3 hours, then add an appropriate amount of Tris aqueous solution, mix and react on a mixer at room temperature for 10 ~30 minutes, the mixture was dialyzed with phosphate buffer solution, and biotinylated rabbit IgG was obtained after dialysis;
[0050] Coating antibody: mix an appropriate amount of biotinylated rabbit IgG with streptavidin magnetic particles, mix and react on a mixer at room temperature for 0.5 to 3 hours, and use magnetic particle protection buffer for the coated magnetic particles After washing, resuspend to 0.2-2 mg / mL to obtain reagent R1. (The coating amount of biotinylated rabbit IgG ranges from 0.3 to 60 μg / mg). The form...
Embodiment 1
[0078] The components of reagent R3 in the kit of this embodiment are shown in the table below:
[0079] Table 8 is the composition of reagent R3 in embodiment 1
[0080]
[0081]
[0082] The detection CV value of the kit prepared using the reagent R3 of Example 1 is less than 3% when performing sample detection. See the table below for details.
[0083] Table 9 is the CV value repeated 10 times for the same sample
[0084]
[0085] In the kits of the following comparative examples, the reagents R1 and R2 are the same, and only the reagent R3 is different.
Embodiment 2
[0100] This embodiment is for the detection of samples with known concentrations. The reagent R1 and reagent R2 for detection are the same, and the reagent R3 is the use of Example 1, no reagent R3 and physiological saline instead of reagent R3. The detection method is the same as the detection method of rheumatoid factor concentration in human serum (plasma). The results are shown in the table below:
[0101] Table 13 is the chemiluminescent photon intensity detected for known samples
[0102]
[0103] As can be seen from Table 13, the detection sensitivity of the kit with reagent R3 is high, and the signal ratio between the concentrations is high; the detection sensitivity and the signal ratio between the concentrations without reagent R3 are low; the use of normal saline The detection sensitivity of the replacement reagent R3 is low, which is not conducive to the detection of low-concentration samples.
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