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A kind of total bile acid detection kit

A technology for detecting kits and total bile acids, which is applied in the direction of determination/inspection of microorganisms, biochemical equipment and methods, etc., can solve the problems of decreased stability of reagent detection, increased blank absorbance, abnormal test results, etc., and improved the reaction curve. , The effect of small fluctuation of the reaction curve and good stability

Active Publication Date: 2022-06-21
山东康华生物医疗科技股份有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, through comparative experiments with a large number of reagents from different manufacturers, our laboratory found that the total bile acid detection kit based on this reaction principle also has certain disadvantages in sample detection: (1) The reagent is not effective in detecting some special samples (especially young children) Serum or plasma), the reagent R1 will react with the sample, causing the reaction curve to drift, thereby increasing the absorbance of the blank, resulting in abnormal test results; (2) As the reagent storage time prolongs, the detection sensitivity of the reagent will decrease, making the factor The K value increases, resulting in a decrease in the detection stability of the reagent

Method used

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  • A kind of total bile acid detection kit
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  • A kind of total bile acid detection kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1-3

[0027] Embodiments 1-3 relate to a total bile acid detection kit with better reaction curve and higher stability, the kit includes reagent R1 and reagent R2, and the components and concentrations of the reagents R1 and R2 are shown in Table 1 :

[0028] Table 1

[0029]

[0030] The total bile acid detection kits described in Examples 1 to 3 are applicable to various types of automatic biochemical analyzers, taking Hitachi 7170 automatic biochemical analyzer as an example.

[0031] Analysis method: rate method;

[0032] Measurement wavelength: main wavelength 405nm, secondary wavelength 660nm;

[0033] Sample: Reagent R1: Reagent R2=3:210:70 (unit ul)

[0034] Operation method: add 3ul sample to 210ul reagent R1, incubate at 37℃ for 5min, add 70ulR2, delay reading for 60 seconds, the reading time is about 180 seconds, the specific operation is shown in Table 2:

[0035] Table 2

[0036]

Embodiment 4

[0037] Observation and evaluation of the reaction curve of embodiment 4

[0038] Evaluation method: by adjusting the reagent R1 to different pH and adding isotonic regulators of different types and concentrations to establish Example 4, Comparative Examples 4-1, 4-2 and 4-3, observe and record the fluctuation of the reaction curve, implement The dosages and specific results of each component of Example 4 and Comparative Examples 4-1, 4-2 and 4-3 are shown in Table 3;

[0039] Evaluation standard: The number of "+" is used to represent the drift or fluctuation of the response curve. The more "+", the more severe the curve drifts, the greater the fluctuation, the worse the curve; on the contrary, the less "+", the more the curve drifts. The smaller it is, the smaller the fluctuation and the smoother the curve.

[0040] table 3

[0041]

[0042] The results in Table 3 show that by adjusting the pH of the reagent R1 and at the same time adding an isotonic adjusting agent of a...

Embodiment 5

[0043] Example 5 Screening test and comparative verification test of stabilizer

[0044]Stabilizer screening: Select one or several combinations of different stabilizer types to establish Example 5 and Comparative Examples 5-1 and 5-2, and perform accelerated destruction at 37°C at the same time, and perform calibration after the same destruction time interval , record the key factor K value. We take K=1000 (combining the technical requirements of many different manufacturers) as the judgment standard. If the value exceeds this value, it means that the reagent does not meet the requirements. Only the comparative effects of the two groups of comparative examples are listed below, but it should be known that the specific comparative examples also include many other combinations of different stabilizers. The formulas of the examples and comparative examples are shown in Table 4. During the accelerated destruction experiment, the K value The changes are shown in Table 5.

[0045...

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Abstract

The present invention relates to the technical field of detection of in vitro diagnostic reagents, in particular to a total bile acid detection kit, comprising reagent R1 and reagent R2, reagent R1 comprising a buffer, a surfactant, a preservative, an isotonic regulator and Thio-NAD; Reagent R2 includes buffer, surfactant, stabilizer, preservative, reduced coenzyme I and 3α-hydroxysteroid dehydrogenase, and the buffer in reagent R1 is glycine buffer, citric acid-sodium citrate buffer, ortho Potassium hydrogen phthalate-sodium hydroxide buffer, PBS buffer, etc.; the isotonic regulator in reagent R1 is NaCl, KCl, MgCl 2 One or more of them; the stabilizers in reagent R2 are TEA, BSA, TCEP, glutathione, mannitol, etc. When the kit of the invention is used to measure the total bile acids of special samples, the reaction curve can be improved, and the long-term storage stability is good.

Description

technical field [0001] The invention relates to the technical field of in vitro diagnostic reagent detection, in particular to a total bile acid detection kit. Background technique [0002] Bile acid is an important component of bile and plays an important role in fat metabolism. Bile acid mainly exists in the enterohepatic circulatory system and plays a certain protective role through recycling. Only a small part of bile acid enters the peripheral circulation and promotes bile acid intestinal tract. The driving force of the hepatic circulation is the transport system of hepatocytes, the absorption and secretion of bile acids into bile, the cholecystokinin-induced gallbladder contraction, the propulsive peristalsis of the small intestine, the active transport of the ileal mucosa, and the inflow of blood into the portal vein. [0003] Total bile acid (TBA) is a general term for a series of substances, including cholic acid (CA), chenodeoxycholic acid (CDCA), deoxycholic acid ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/32
CPCC12Q1/32
Inventor 杨致亭秦冬立孙异凡吴国才戴艳霞
Owner 山东康华生物医疗科技股份有限公司
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