Immunoturbidimetric detection reagent and method for serum amyloid protein A

An immunoturbidimetric and serum starch technology, which is applied in the field of medical immunodiagnosis, can solve the problems of unfavorable small and medium-sized hospitals, reduced test accuracy, and inconvenient use, and achieves the effects of shortening test time, simplifying operation, and improving speed

Inactive Publication Date: 2018-03-30
SUZHOU KANGHESHUN MEDICAL TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The operation of dilution and mixing can be automatically operated by machines on automatic biochemical analyzers (commonly used instrument platforms in tertiary hospitals), but on small semi-automatic platforms, especially POCT portable platforms (hospitals below the third level and medical services at the county and township level) The test performed on the unit) needs to be manually implemented by the operator, which brings additional operation steps and introduces more operational errors, which reduces the accuracy of the test and is not conducive to the promotion and implementation of the test in small and medium-sized hospitals.
Another disadvantage of the SAA latex-enhanced immune turbidimetric reagents on the existing market is that it can only measure the SAA content in serum or plasma samples, and the whole blood samples must be pre-treated to extract serum, which is time-consuming and laborious. In clinical practice Very inconvenient to use

Method used

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  • Immunoturbidimetric detection reagent and method for serum amyloid protein A
  • Immunoturbidimetric detection reagent and method for serum amyloid protein A
  • Immunoturbidimetric detection reagent and method for serum amyloid protein A

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] 1) Preparation of latex reagent

[0042] The surface carboxylated microspheres with a particle size of 60nm were diluted to 1% in MES (50mM, pH=6.2) buffer solution and stirred at room temperature. Put in EDC solid powder, keep the molar ratio of EDC and carboxyl group at 10:1, stir for 2 hours and then centrifuge at 10000RPM for 10 minutes. The latex was washed twice with MES (50mM, pH=6.2) buffer solution and resuspended. Add SAA antibody, the ratio of antibody mass to latex mass is 0.025:1, and stir for 2 hours. Then centrifuge at 10,000 RPM for 15 minutes, and discard the supernatant. Latex was resuspended in storage solution (50mM MOPSO+0.5%BSA+0.1%ProClin 300+0.5%Triton X-100+0.5%trehalose+0.9%NaCl, pH7.0) and ultrasonically dispersed, diluted to a latex concentration of About 0.5% is reserved.

[0043] 2) Preparation of standard products

[0044] The commercially available SAA antigen standard was prepared into a series of standard products with 50mM PBS, th...

Embodiment 2

[0050] 1) Preparation of latex reagent

[0051] The surface carboxylated microspheres with a particle size of 60nm were diluted to 1% in MES (50mM, pH=6.2) buffer solution and stirred at room temperature. Put in EDC solid powder, keep the molar ratio of EDC and carboxyl group at 10:1, stir for 2 hours and then centrifuge at 10000RPM for 10 minutes. The latex was washed twice with MES (50mM, pH=6.2) buffer solution and resuspended. Add SAA antibody, the ratio of antibody mass to latex mass is 0.05:1, and stir for 2 hours. Then centrifuge at 10,000 RPM for 15 minutes, and discard the supernatant. The latex was resuspended in storage solution (50mM PBS+0.5%BSA+0.1%ProClin 300+0.5%Triton X-100+0.5% trehalose, pH 7.0) and ultrasonically dispersed, diluted to a latex concentration of about 0.5% for later use.

[0052] 2) Preparation of standard products

[0053] The commercially available SAA antigen standard was prepared into a series of standard products with 50mM PBS, the con...

Embodiment 3

[0059] Analytical sensitivity of reagents (blank limit)

[0060] Using the reagents in Example 1, select the zero-value standard as a blank sample test, and test the absorbance value on an automatic biochemical analyzer (Mindray BS-400) at a temperature of 37°C, a wavelength of 546nm, and an optical path of 1cm. Repeat the test 20 times, calculate the mean (X) and standard deviation (SD) of the 20 test results according to the calibration curve established in Example 1, and calculate X+2SD as the analytical sensitivity of the reagent. The test results are shown in Table 1, showing that the analytical sensitivity of the reagent is 0.58 mg / L. The clinical reference value for detection of serum amyloid A (SAA) is about 10 mg / L, and the sensitivity of the reagent of the present invention is an order of magnitude smaller than the reference value, which fully meets the needs of use.

[0061] Table 1. The reagent analysis sensitivity test result of embodiment 1

[0062] Ex...

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Abstract

The invention discloses a latex-enhanced immune turbidimetric detection reagent for serum amyloid A (SAA‑Serum Amyloid A). The reagent is a single reagent, and the main component of the reagent is latex particles labeled with SAA antibody, and also includes buffer solution, surfactant, salt, stabilizer, suspending agent and preservative. The invention also discloses a method for using the reagent to detect the concentration of serum amyloid A (SAA) in blood samples by the principle of transmission or scattering turbidimetry. The invention adopts a single reagent, is simple to operate, does not need to be mixed, has high sensitivity, and has a wide linear range, and can directly measure various samples such as whole blood, serum, and plasma, and can be widely used in various transmission or scattering analyzers, including ordinary biochemical analyzers , specific protein analyzers, etc.

Description

technical field [0001] The invention belongs to the field of medical immunodiagnosis, and in particular relates to the preparation and application of a latex-enhanced immune turbidimetric detection reagent for serum amyloid A (SAA). Background technique [0002] Serum amyloid A (SAA), an acute phase protein associated with high-density lipoprotein (HDL), is mainly produced in the liver and is regulated by interleukin 1 (IL-1), interleukin 6 (IL-6), tumor Regulation of cytokines such as necrosis factor (TNF-alpha). When inflammation occurs or infection with bacteria or viruses occurs, the concentration of SAA in the blood can rise to more than 1000 times the normal value within a few hours, which is a very important infection index in clinical application (Maury CPJ, Clinical science , 1985, 68:233-238). Compared with CRP (C-reactive protein), another important infection indicator in clinical application, SAA is more sensitive to viral infection, with a larger increase an...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N21/31G01N21/47G01N21/59
CPCG01N33/68G01N21/31G01N21/47G01N21/59
Inventor 刘向晖陈胜胜王明
Owner SUZHOU KANGHESHUN MEDICAL TECH
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