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Preparation and detection method of hepatobiliary acid determination kit

A kit and reagent technology, applied in the field of pharmaceutical reagents, can solve the problems of high cost, long measurement time, errors, etc.

Pending Publication Date: 2020-09-25
JILIN GETEIN BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, the enzyme-linked immunosorbent assay has high detection accuracy, but the operation process is slightly complicated, the measurement time is long, and there are many influencing factors, and it is not suitable for the detection of emergency samples; the automatic operation speed of the chemiluminescence immunoassay is fast, but the cost High, expensive equipment, poor stability, and cannot be popularized in middle and primary hospitals and medical institutions; Immunoturbidimetric assay, when the amount of antigen or antibody is too large, soluble complexes may appear, causing errors, and are easily affected by lipemia

Method used

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  • Preparation and detection method of hepatobiliary acid determination kit
  • Preparation and detection method of hepatobiliary acid determination kit
  • Preparation and detection method of hepatobiliary acid determination kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1 Preparation of hepatic bile acid detection kit

[0033] The hepatic bile acid detection kit prepared according to the present invention comprises independently packaged reagent R1 and reagent R2, and the reagent R1 comprises components of the following concentrations:

[0034] Glycine buffer: 100mmol / L;

[0035] Sodium chloride: 9g / L;

[0036] Triton X-100: 0.5g / L;

[0037] PEG20000: 20g / L

[0038] PC300: 1g / L;

[0039] Described reagent R2 comprises the component of following concentration:

[0040] Glycine buffer: 100mmol / L;

[0041] Antibody latex particles: 35mg / L;

[0042] Sucrose: 5g / L;

[0043] PC300: 1g / L;

[0044] Preferably, the preparation steps of the antibody latex particles in the reagent R2 are:

[0045] Preparation: prepare 220nm polystyrene latex particles, and dilute the latex particles with 5mM / L 2-(N-morpholine)ethanesulfonic acid at pH 7.0 to a final concentration of 1.5% w / v to obtain a latex particle solution; Goat polyclonal a...

Embodiment 2

[0049] Embodiment 2 Analysis of the accuracy of the kit of the present invention

[0050] Test kit: the kit of Example 1, the hepatocholic acid assay kit of the comparative example (produced by Beijing Jiuqiang Biotechnology Co., Ltd.)

[0051] Test instrument: Hitachi 7100 automatic biochemical analyzer

[0052] The specific operation is to set parameters: test method: two-point end point method; main wavelength: 600nm; photometry point: 18-34; sample size: 6; reagent volume R1: R2=150: 50; calibration method: spline

[0053] Setting of the standard curve: Put the standard substances of known concentration, namely 5, 10, 20, 40, 80 mg / L in the calibration plate, set the position and concentration of the calibration substance, and obtain the following data:

[0054] Table 1 (Example 1)

[0055]

[0056] Table 2 (comparative example)

[0057]

[0058] 40 cases of clinical samples with different concentrations were tested in parallel with the control, and the following ...

Embodiment 3

[0064] Embodiment 3 Precision analysis of the kit of the present invention

[0065] Test kit: the kit of Example 1, the hepatocholic acid assay kit of the comparative example (produced by Beijing Jiuqiang Biotechnology Co., Ltd.)

[0066] Test instrument: Hitachi 7100 automatic biochemical analyzer

[0067] The specific operation is to set parameters: test method: two-point end point method; main wavelength: 600nm; photometry point: 18-34; sample size: 6; reagent volume R1: R2=150: 50; calibration method: spline

[0068] After calibration, the same serum sample was measured for 10 times, and the average value and SD value were obtained from the concentration results of 10 times of detection, and the CV value was calculated. The results are shown in Table 5:

[0069]

[0070] As can be seen from the data in Table 5, the repeatability of the measurement results of the same sample in Example 1 is almost the same as that of the comparative example, indicating that this reagen...

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Abstract

The invention aims to provide preparation of a hepatobiliary acid detection kit. The method comprises the steps of: adding the prepared antibody latex particles into a reagent R2; and preparing the reagent R1 and the reagent R2 which are independently packaged, detecting the hepatobiliary acid in a serum sample by adopting a latex enhanced turbidimetric immunoassay method, and therefore the detection is rapid and accurate, the accuracy is high, the specificity is strong, the precision is good, the cost is relatively low, the detection sample amount is small, and a small amount of samples and emergency treatment samples can be determined.

Description

technical field [0001] The invention belongs to the technical field of medical reagents. Background technique [0002] Serum Cholyglycine (CG) is one of the conjugated bile acids formed by the combination of bile acid and glycine. In liver cells, cholesterol is transformed into primary bile acid through complex enzymatic reactions. Among them are cholic acid (CA) and chenodeoxycholic acid (CD-CA). There are three hydroxyl groups (C3, C7, C12) on the steroid core of cholic acid, and the hydroxyl group at the end of the side chain is combined with glycine through a peptide bond, with a molecular weight of 462u. [0003] The normal metabolic pathway of CG is the entero-hepatic circulation. CG is synthesized by liver cells, discharged into the gallbladder through the capillaries and bile ducts, and enters the duodenum with bile to help digest food. 95% of bile acid is reabsorbed in the terminal ileum, and then returned to the liver through the portal vein, where it is taken up...

Claims

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Application Information

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IPC IPC(8): G01N33/543G01N33/53
CPCG01N33/54313G01N33/5308
Inventor 陈凯丽徐李鹏刘玲
Owner JILIN GETEIN BIOTECH CO LTD