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Stable high-interference-resistance direct bilirubin (oxidase method) detection reagent and detection method

A technology for detecting reagents and bilirubin, applied in the measurement of color/spectral properties, etc., can solve the problems of poor anti-interference ability and stability, difficult to popularize and apply, poor sensitivity and anti-hemolysis ability, etc., and achieve good accuracy and stability. , The effect of strong anti-interference and improved anti-interference ability

Inactive Publication Date: 2019-07-09
济南宇鑫生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, the detection methods of direct bilirubin include diazo reagent method, high performance liquid chromatography, bilirubin oxidase method, etc. Among them, the diazo reagent method is simple to operate, but the method has poor sensitivity and anti-hemolysis ability; Liquid chromatography has good sensitivity and specificity, but requires special instruments and is difficult to popularize and apply; while bilirubin oxidase method has high sensitivity and is not affected by hemolysis, but has poor anti-interference ability and stability

Method used

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  • Stable high-interference-resistance direct bilirubin (oxidase method) detection reagent and detection method
  • Stable high-interference-resistance direct bilirubin (oxidase method) detection reagent and detection method
  • Stable high-interference-resistance direct bilirubin (oxidase method) detection reagent and detection method

Examples

Experimental program
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Effect test

Embodiment 1

[0053] Detection reagents for serum direct bilirubin, including reagent R1 and reagent R2:

[0054] 1) The composition of its R1 is:

[0055] MES (2-(N-morpholine)ethanesulfonic acid) buffer solution (pH=4.5, 25°C) ·················· 50mmol / L,

[0056] sodium fluoride ······················································· 2mmol / L,

[0057] NAC (N-acetylcysteine) ·········································· 0.2 mmol / L,

[0058] Sodium 4-toluenesulfonate ················································· 40mmol / L,

[0059] Lipase ······················································· 15KU / L,

[0060] ascorbate oxidase ··············································· 8 KU / L,

[0061] α-cyclodextrin ····················································· 1.5g / L,

[0062] sucrose ························································· 40g / L,

[0063] Trehalose ······················································· 10g / L,

[0064] PEG-6000 ···························...

Embodiment 2

[0078] 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 use the reagent obtained in Example 1, and the direct bilirubin (oxidase method) reagent that is common and recognized in the market to compare and measure the content of direct 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%.

[0079] Depend on Figure 4 It can be seen that in the sample, when hemoglobin≤90mg / dL, chylomicrons≤2700 turbidity units, glucose≤500mmol / L, ascorbic acid≤10mg / dL, urea≤17mmol / L, and uric acid≤2μmol / L, the reagent of Example 1 There was no significant interference in the test results. However, the reagents in the control group were...

Embodiment 3

[0081] Correlation experiment: using the formula in Example 1 to prepare reagents, and conducting a control test with a company’s direct bilirubin (oxidase method) detection kit approved by the State Food and Drug Administration, which is common in the market, and testing 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 As shown), the test results show that the correlation coefficient of the two kits is 0.9996, which shows that there is a great correlation between the two.

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Abstract

The invention relates to the field of serum direct bilirubin detection technologies, in particular to a serum direct bilirubin (oxidase method) detection reagent. A reagent R1 contains a buffer solution, sodium fluoride, NAC(N-acetylcysteine), sodium 4-methylbenzenesulfonate, lipase, ascorbic acid oxidase, alpha-cyclodextrin, sucrose, trehalose, PEG-6000, fatty alcohol polyoxyethylene ether (AEO-9) and a preservative; and a reagent R2 contains a buffer solution, bilirubin oxidase, sucrose, trehalose, PEG-6000, fatty alcohol polyoxyethylene ether (AEO-9) and a preservative. Through the reagent,the specificity and accuracy of direct bilirubin testing are obviously improved, turbidity of a reaction system can be prevented, a reaction is promoted, and the stability and interference resistanceof the reagent are further improved.

Description

technical field [0001] The invention relates to the technical field of serum direct bilirubin detection, in particular to a serum direct bilirubin detection reagent and a detection method using the detection reagent. Background technique [0002] Bilirubin is formed by deferrization of heme released after red blood cells are destroyed. 80% of bilirubin comes from spleen and liver, and the rest comes from bone marrow. Bilirubin exists in three forms, indirect or unconjugated bilirubin, conjugated bilirubin, and delta bilirubin. Among them, combined bilirubin and delta bilirubin can directly react with diazo reagents, which are called direct bilirubin. [0003] Various types of hepatitis, liver cirrhosis, and obstructive jaundice can be seen to increase the content of direct bilirubin, and it is more sensitive than transaminase, especially suitable for the evaluation of the prognosis of chronic hepatitis and the diagnosis of early liver cirrhosis. Serum direct bilirubin leve...

Claims

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Application Information

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IPC IPC(8): G01N21/31
CPCG01N21/31
Inventor 李志明胡晓飞李道伟李贵雨
Owner 济南宇鑫生物科技有限公司
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