CN-122017220-A - Method for simultaneously improving sensitivity and cross-contamination resistance of latex turbidimetry kit and application thereof

Abstract

The invention discloses a method for simultaneously improving sensitivity and cross-contamination resistance of a latex turbidimetric kit and application thereof, and belongs to the technical field of biology. The invention discloses a method for simultaneously improving the sensitivity and the cross-contamination resistance of a latex turbidimetric kit, wherein latex microspheres used in a sensitized latex solution are low-mass-density latex microspheres, and the sensitized latex solution contains prednisone. The method improves the sensitivity by 39-46%, prolongs the airborne service time by 3-10 times after the reagent is cross-polluted, prolongs the scrapping time from 2-3 days to 7-30 days and ensures the stability, thereby meeting the basic use requirement.

Inventors

• Liu Xiangdai
• XUE YUTING
• BAI JING
• WANG ZHAO
• JIANG XIN

Assignees

• 首都医科大学附属北京同仁医院
• 柏荣诊断产品（上海）有限公司

Dates

Publication Date

20260512

Application Date

20260203

Claims (5)

1. 1. A method for simultaneously improving sensitivity and cross-contamination resistance of a latex turbidimetric kit is characterized in that the latex turbidimetric kit comprises a reagent R1 and a reagent R2, wherein the reagent R1 is a reaction buffer solution, and the reagent R2 is a sensitization latex solution; the latex microspheres used in the sensitization latex solution are low-mass-density latex microspheres, and the sensitization latex solution contains prednisone; the mass density of the low mass density latex microsphere is 0.92-0.96g/ml; the temperature sensitive immunosuppressant prednisone is used in an amount of 0.20-2.00mg in 1L of the sensitization latex solution.
2. 2. A method of simultaneously increasing the sensitivity and cross-contamination resistance of a latex turbidimetric kit according to claim 1, said low mass density latex microspheres having a mass density of 0.94g/ml; the prednisone is used in an amount of 0.8-1.4mg in 1L of the sensitization latex solution.
3. 3. The method for simultaneously improving sensitivity and cross-contamination resistance of a latex turbidimetric kit according to claim 1, wherein the low-mass-density latex microspheres are hollow latex microspheres prepared by a seed method or other methods, the seeds are polymers with mass density lower than 1.03g/ml, and the polymers are polyethylene or polypropylene.
4. 4. The method for simultaneously improving the sensitivity and the cross-contamination resistance of a latex turbidimetric kit according to claim 1, wherein on a full-automatic biochemical or specific protein analyzer, a sample and a reagent R1 are mixed and incubated for 1 to 5 minutes, then a reagent R2 sensitized latex solution is added for carrying out antigen-antibody latex reaction, a delta signal value is calculated by a two-point endpoint method, and after the sensitized latex solution is added into a mixed reaction solution of the reagent R1 and the sample, a first optical signal acquisition is carried out, and after an immune reaction is carried out for 2 to 5 minutes, a second optical signal acquisition is carried out.
5. 5. Use of the method according to any one of claims 1-4 for simultaneously increasing the sensitivity and cross-contamination resistance of latex turbidimetric kits.

Description

Method for simultaneously improving sensitivity and cross-contamination resistance of latex turbidimetry kit and application thereof Technical Field The invention relates to the field of biotechnology, in particular to a method for simultaneously improving sensitivity and cross-contamination resistance of a latex turbidimetry kit and application thereof. Background Latex immunoturbidimetry is a mainstream method in the fields of biochemistry and specific protein detection due to the popularization of equipment (various biochemical transmission, specific protein scattering and transillumination fusion analyzers), rapid detection speed and quantification (quantitative results are usually given in 8-10 minutes, even 1-2 minutes in POCT), and high detection throughput (from 50T/hour of a mini-machine to 2000T/hour of a large-scale equipment). The main performances of the latex immunoturbidimetry kit comprise sensitivity, linearity, antigen surplus safety region, specificity, reagent stability and the like, and a plurality of enterprises and research institutions respectively invest resources in the aspects of the performances for upgrading and optimizing. For example, in the aspect of reagent sensitivity, CN 201811426490.6 is a method for improving the detection sensitivity of a latex turbidimetric reagent, and research and exploration are carried out in the aspect of improving the sensitivity of a kit by introducing an amino acid bracket to change the steric hindrance of two Fab of an antibody. CN202010813669.8 is a polystyrene microsphere for improving sensitivity of latex turbidimetry and its preparation method, then research and exploration are conducted from the aspect of increasing the number and length of carboxyl chains on the surface of polystyrene microsphere. The CN201410401284.5 anti-cyclic citrullinated peptide antibody detection kit increases the variation intensity of scattered light and transmitted light from high refractive index and chemical inertia, so that research and exploration are conducted in the aspect of improving the detection sensitivity. CN202510645838.4 is a method for improving sensitivity of latex turbidimetric immunoassay reagent, CN102759631B is a latex enhanced turbidimetric immunoassay kit for quantitatively detecting procalcitonin PCT, and CN108872616B is an immune latex turbidimetry kit for detecting NGAL based on single-particle-size latex particles, and a biotin-streptavidin binding system is introduced to improve detection sensitivity of the latex turbidimetric reagent. Various researches of a plurality of enterprises and research institutions aim to improve the detection sensitivity of the latex turbidimetric kit, so that a better detection effect is achieved, or more items are detected. But there is rarely a study to control the reaction rate of different sections of the reaction curve and thus the sensitivity of the reagents and their ability to resist cross-contamination. Therefore, it is a need for those skilled in the art to provide a method and application for improving the sensitivity and cross-contamination resistance of latex turbidimetric kits. Disclosure of Invention In view of the above, the invention provides a method and application for simultaneously improving the sensitivity and the cross-contamination resistance of a latex turbidimetric kit. In order to achieve the above purpose, the present invention adopts the following technical scheme: It is currently rare to control the reaction rate at different sections of the reaction curve and thus the reagent sensitivity. Taking Hitachi 7180,3500 as an example, a typical latex turbidimetric reaction curve is shown in FIG. 1, with the abscissa representing the reaction time, counting at 1 to 34 photometric time points, and the ordinate representing the reaction signal value (transmission is absorbance ABS, and scattering is light intensity INT). The sample is added to buffer and incubation is typically performed within 1 to 16 photometric points, typically between 16 and 17 photometric points, and the instrument is added with an antibody-labeled priming latex, and it can be seen that from 17 photometric points, an antigen-antibody latex immune complex is gradually formed, and the turbidity increases with an increase in signal value, but the immune reaction does not reach the end of the reaction within 5 minutes of the usual 17 to 34 photometric points. The parameters are two-point endpoint method, 18-34 read points means that the first light measuring point is 18 light measuring points after the sensitized latex is added, and the second light measuring point is 34 light measuring points at the tail. When the immune response does not reach the end of the reaction, the rate of the reaction and the shape of the reaction curve will have an effect on the delta signal value calculation. 18-34 of the read points of the wafer, the calculation mode is delta signal value= (34 light measurement point signal value+33 ligh