A kit for the detection of tetanus antibodies in human plasma by latex-enhanced immune turbidimetry
An immunoturbidimetric and latex-enhanced technology, used in immunoassays, biological tests, measuring devices, etc., can solve the problems of inaccurate measurement of tetanus antibody titer, difference between batches and plates, and short detection time.
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Embodiment 1
[0095] The preparation of the latex particle that embodiment 1 is coated with tetanus antigen
[0096] The polystyrene latex particles used in the kit of the present invention are all self-produced, by the polymerization method of soap-free emulsion (do not add emulsifier or add a small amount of emulsifier), with persulfate (potassium persulfate, sodium persulfate, persulfate ammonium, etc.) as an initiator to initiate the copolymerization of styrene and vinyl carboxylic acid functional monomers (methacrylic acid, acrylic acid, maleic acid, etc.) to prepare polystyrene microspheres rich in carboxyl groups on the surface. By adjusting parameters such as water-oil ratio, functional monomer ratio, and reaction temperature, it is possible to prepare microspheres with an adjustable particle size in the range of 50nm to 350nm and a controllable carboxyl density in the range of 0-400μmol / g.
[0097] Get particle diameter respectively and be each 6.47mL of the polystyrene latex parti...
Embodiment 2
[0104] The particle diameter of the latex particle of embodiment 2 coating tetanus antigen and the linear correlation coefficient determination of reactivity
[0105] Get the latex particles coated with tetanus antigen with a particle diameter of 180nm, 200nm, 257nm, 350nm, 400nm prepared in Example 1, the concentration is 0.19wt% by mass, test the 5-point reactivity of the calibrator respectively, and fit a straight line .
[0106] Experimental results such as figure 1 As shown, the linear correlation coefficients between the particle size and reactivity of the tetanus antigen-coated latex particles all reach above 0.95. However, under the condition of 180nm particle size, the reaction degree is obviously low, and the linear correlation coefficient is 0.96535. When the latex particle size of the antigen is 180nm and 400nm, it cannot be used for the detection of tetanus antibody. And when the particle size of the latex particles coated with tetanus antigen is 200nm-350nm, t...
Embodiment 3
[0107] The linear correlation coefficient determination of different buffer solutions and reactivity in the embodiment 3 reagent R1
[0108] Select Tris buffer (50mM, pH8.5), phosphate buffer (50mM, pH7.2) glycine buffer (50mM, pH6.5), add 1g / LBSA and 1M / LNaCl respectively, and test as reagent R1. Take latex particles coated with tetanus antigen with a particle size of 257nm, the concentration is 0.19wt% by mass, test the 5-point reactivity of the calibrator, and fit a straight line.
[0109] Experimental results such as figure 2 As shown, the linear correlation coefficients all reached more than 0.98, especially the R1 solution prepared with Tris buffer, the linear correlation coefficient reached 0.99855.
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