Detection kit for cystine protease inhibitor C

A cystine protease and detection kit technology, applied in the field of medical immunology, can solve the problems of increasing antibody dosage and production cost, weak anti-interference ability, low accuracy, etc. The effect of strong interference ability and lower production cost

Active Publication Date: 2012-02-15
NINGBO MEDICAL SYSTEM BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, when the cystatin C antibody is coated on the latex, physical adsorption and chemical coupling methods can generally be used. The stability of the sensitized latex particles obtained by the physical adsorption method is worse than that of the chemical coupling method, and the antibody is easily released from the latex. and the latex obtained by physical adsorption may be interfered by rheumatoid factor (RF) and heterophilic antibodies. IgM and IgG type RF can directly bind to the Fc segment of the antibody coated on the latex , resulting in false positive or falsely elevated test results
Heterophilic antibodies can bind to the Fc region of rodent IgG, so that heterophilic antibodies can bind to latex-coated antibodies, resulting in false positive or falsely elevated test results
Most of the chemical coupling methods currently used are random coupling methods, because randomly coupled antibodies are randomly coupled to different parts of the antibody, which will lead to the loss of binding force of the antibody; therefore, complete random coupling will lose most of the binding force of the antibody and increase Antibody dosage and production cost
At the same time, if the sensitized latex particles obtained by the above method are used in the detection kit, it will cause the defects of poor specificity, low accuracy, weak anti-interference ability or high production cost.

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  • Detection kit for cystine protease inhibitor C
  • Detection kit for cystine protease inhibitor C

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1 Preparation of Cystatin C Detection Kit

[0028] The kit of the present invention relates to the main raw materials of reagents as follows:

[0029] 1. Cystatin C antibody: titer determined by indirect ELISA method is 1:100000.

[0030] 2. Latex: The present invention only exemplarily uses polystyrene latex particles with a diameter of 100-200 nm and carboxyl groups to carry out experiments.

[0031] The preparation of the main reagent of this embodiment is as follows:

[0032] Reagent R1: ammonium chloride buffer containing 1.5% PEG6000 (polyethylene glycol 6000), 0.8% NaCl. The reagent is a colorless transparent solution.

[0033] Reagent R2: Latex particles were sensitized with anti-cystatin C antibody. The reagent is a milky white solution. Specific steps are as follows:

[0034] 1. Take 1ml (100mg / ml) latex, wash 3 times with 0.1M (mol / L), pH5.0 MES solution (2-morpholineethanesulfonic acid buffer), and ultrasonically disperse;

[0035] 2. Add 0.2m...

Embodiment 2

[0044] Example 2 Determination of Cystatin C

[0045] Detection tool: Hitachi 7060 automatic analyzer.

[0046] Analysis method: two-point endpoint method; main wavelength: 546nm, secondary wavelength: 700nm: sample volume: 3.0ul; R1: 250ul; R2: 50ul; calibration method: spline function Spline; reaction direction: rising; measurement temperature: 37℃ ;After mixing the sample with R1, read the absorbance A at 30 seconds 1 ;Add R2 at 5 minutes, read absorbance A at 5 minutes 2 . The absorbance of the reaction was calculated as A 2 with A 1 calibration difference.

[0047] Calculation method: multi-point calibration, using spline function as the calculation mode, and making a dose / response curve according to the absorbance and reference serum value, and the sample content can be calculated on the dose / response curve according to its absorbance value.

[0048] Reference range: 0.55~1.05mg / L

Embodiment 3

[0049] Example 3 Testing of various performance indicators of the cystatin C detection kit

[0050] 1. Sensitivity test

[0051] Measure the sample of 6 kinds of different cystatin C contents, each sample measures 10 times, by calculating mean value and SD value (standard deviation value), as seen from Table 1, the sensitivity of detection kit of the present invention is 0.08mg / L.

[0052] Table 1

[0053] CYS-C (mg / L)

[0054] 2. High value linear determination

[0055] Measure 10 samples of different cystatin C content, each sample is measured 2 times, as seen from Table 2, the highest detection range of the detection kit of the present invention can reach 8mg / L, judge basis r 2 ≥0.990.

[0056] Table 2

[0057] Theoretical value of CYS-C (mg / L)

[0058] 3. Precision test

[0059] Two human serum samples with different cystatin C contents were used to determine the intra-assay precision of the detection kit of the present invention. Use 3 batche...

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Abstract

The invention discloses a detection kit for a cystine protease inhibitor C. The kit comprises a reagent R1 and a reagent R2, wherein the R1 is a proper buffer, and the reagent R2 is formed through placing polystyrene latex particles sensitized by cystine protease inhibitor C antibodies in a proper buffer. The kit of the present invention has the advantages of high sensitivity, good specificity, good accuracy, strong anti-interference capability, and low production cost.

Description

technical field [0001] The invention belongs to the field of medical immunology, and relates to an immunological detection reagent, and further, the invention relates to a cystine protease inhibitor C detection kit. Background technique [0002] Creatinine, so far widely used as a marker of glomerular filtration (GFR), is a small molecular substance produced by muscle, so the rate of creatinine formation and secretion is closely related to the muscle content in the body. It is well known that within a population, individuals have relatively different muscle mass. Older adults have less muscle mass than young adults. Many patients experience loss of muscle mass over the course of their disease. Children also have different muscle mass than adults. Men The average muscle mass of women is higher than that of women. When serum creatinine concentration is used as a marker of glomerular filtration rate, the detected serum creatinine value is 50% lower than the actual value. Furt...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/536G01N33/531
Inventor 邹炳德夏佳音
Owner NINGBO MEDICAL SYSTEM BIOTECHNOLOGY CO LTD
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