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Stable total bilirubin (oxidase method) detection reagent with strong anti-interference ability and detection method

A technology for total bilirubin and detection reagents, applied in the field of serum total bilirubin detection reagents, can solve problems such as poor anti-interference ability and stability, poor sensitivity and anti-hemolysis ability, and achieve good accuracy and stability, enhanced Anti-interference ability, enhanced stability and effect of anti-interference ability

Inactive Publication Date: 2019-09-24
济南煊赫生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] At present, the detection methods of total bilirubin include diazo reagent method, high performance liquid chromatography, bilirubin oxidase method, etc., among which the diazo reagent method is simple to operate, but the method has poor sensitivity and anti-hemolysis ability; Phase chromatography has high sensitivity and specificity, but requires special instruments; while bilirubin oxidase method has very good specificity and sensitivity, but poor anti-interference ability and stability

Method used

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  • Stable total bilirubin (oxidase method) detection reagent with strong anti-interference ability and detection method
  • Stable total bilirubin (oxidase method) detection reagent with strong anti-interference ability and detection method
  • Stable total bilirubin (oxidase method) detection reagent with strong anti-interference ability and detection method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Serum total bilirubin detection reagents, including reagent R1 and reagent R2:

[0047] 1) The composition of its R1 is:

[0048] PIPES (1,4-piperazine diethanesulfonic acid) buffer solution (pH=7.2, 25°C) ······· 100mmol / L,

[0049] Lipase ············································ 20KU / L,

[0050] ascorbate oxidase ···································· 10 KU / L,

[0051] α-cyclodextrin ·········································· 2g / L,

[0052] sucrose ··········································· ·· · 50g / L,

[0053] Trehalose ············································ 20g / L,

[0054] PEG-6000 ·········································· 4g / L,

[0055] Castor Oil Polyoxyethylene Ether (EL-20) ··························· 2g / L,

[0056] Preservative Proclin300 ·································· 0.2mL / L;

[0057] 2) The components of reagent R2 are:

[0058] PIPES (1,4-piperazine diethanesulfonic acid) buffer solution (pH=7.2, 25°C) ···· ·· 100mmol / L,...

Embodiment 2

[0068] Interference test: Take fresh mixed serum, divide it into 2 equal parts, then divide each equal part into 7 equal parts, add different interfering substances, so that the concentration in the serum reaches Figure 4 requirements. Then respectively use the reagent obtained in Example 1, and the total bilirubin (oxidase method) reagent that is common and recognized in the market to compare and measure the content of total bilirubin in serum at the same time. See the test results Figure 4 . Relative deviation (%) = (measuring mean value of interference samples - measuring mean value of control samples) / measured mean value of control samples × 100%.

[0069] Depend on Figure 4 It can be seen that in the sample, when hemoglobin≤100mg / dL, chylomicrons≤3000 turbidity units, glucose≤555mmol / L, ascorbic acid≤10mg / dL, urea≤18mmol / L, and uric acid≤2μmol / L, the reagent of Example 1 There was no significant interference in the test results. However, the reagents of the contro...

Embodiment 3

[0071] Correlation experiment: use the formula of Example 1 to prepare reagents, and conduct a control test with a company's total bilirubin (oxidase method) detection kit approved by the State Food and Drug Administration, which is common in the market, and detect 20 clinical sera at the same time samples, the test results are as follows Figure 5 shown. And obtained the correlation curve of the two reagents (such as figure 1 Shown), the test results show that the correlation coefficient of the two kits is 0.9994, indicating that there is a great correlation between the two.

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Abstract

The invention relates to the technical field of total bilirubin detection, especially to a serum total bilirubin (oxidase method) detection reagent. A reagent R1 contains a buffer solution, lipase, ascorbic acid oxidase, alpha-cyclodextrin, sucrose, trehalose, PEG-6000, castor oil polyoxyethylene ether (EL-20) and a preservative. A reagent R2 contains a buffer solution, bilirubin oxidase, sucrose, trehalose, PEG-6000, castor oil polyoxyethylene ether (EL-20) and a preservative. The interference of lipid turbidity and ascorbic acid oxidase can be effectively avoided and the anti-interference ability of the reagent can be greatly enhanced by optimizing a reaction system and adding the lipase, the alpha-cyclodextrin and the ascorbic acid oxidase. The adoption of the novel PIPES(piperazine-1,4-diethylsulfonic acid) buffer solution and the addition of stabilizers such as the sucrose, the trehalose, the polyethylene glycol 6000 can effectively improve the stability of the reagent. Moreover, the addition of the optimal novel non-ionic surfactant castor oil polyoxyethylene ether (EL-20) can prevent the reaction system from turbidity and further improve the stability and the anti-interference ability of the reagent.

Description

technical field [0001] The invention relates to the technical field of serum total bilirubin detection, in particular to a serum total bilirubin detection reagent and a detection method using the detection reagent. Background technique [0002] Bilirubin is formed by the deferrification of heme released after red blood cells are destroyed. 80% of the bilirubin comes from the spleen and liver, and the rest comes from the bone marrow. There are three forms of bilirubin, indirect bilirubin or unconjugated bilirubin, conjugated bilirubin, delta bilirubin, the three are collectively referred to as total bilirubin. [0003] The content of total bilirubin increases, and the content of direct bilirubin and indirect bilirubin also increases, which is common in hepatocellular jaundice, such as acute jaundice hepatitis, severe hepatitis, chronic liver, cirrhosis, etc.; total bilirubin and direct bilirubin Increased bilirubin content is seen in obstructive jaundice, such as biliary st...

Claims

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Application Information

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IPC IPC(8): G01N21/31G01N33/72G01N35/00
CPCG01N21/31G01N33/728G01N35/00
Inventor 李志明李贵雨张丹罗维晓
Owner 济南煊赫生物科技有限公司
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