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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 detection stability of reagents, increased factor K value, increased blank absorbance, etc., and achieves improvement. Response curve, good stabilization effect, effect of reducing fluctuation

Active Publication Date: 2018-06-29
山东康华生物医疗科技股份有限公司
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  • Description
  • Claims
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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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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1-3

[0027] Examples 1-3 respectively relate to a total bile acid detection kit with better response curve and higher stability. The kit includes reagent R1 and reagent R2. 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 1-3 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°C for 5 minutes, add 70ulR2, delay reading for 60 seconds, and the reading time is about 180 seconds. The specific operation is shown in Table 2:

[0035] Table 2

[0036]

Embodiment 4

[0037] The observation evaluation of embodiment 4 response curve

[0038] Evaluation method: by adjusting reagent R1 to different pHs and adding different types and concentrations of isotonic regulators to establish Example 4, Comparative Examples 4-1, 4-2 and 4-3, observe and record the fluctuations in the reaction curves, implement Example 4 and comparative example 4-1, the consumption of each component of 4-2 and 4-3 and concrete result are shown in Table 3;

[0039] Evaluation criteria: Use the number of "+" to indicate the drift or fluctuation of the response curve. The more "+", the more severe the curve drift, the greater the fluctuation, the worse the curve; on the contrary, the less "+", it means the curve drift The smaller the value, the smaller the fluctuation and the smoother the curve.

[0040] table 3

[0041]

[0042] The results in Table 3 show that in the present invention, by adjusting the pH of the reagent R1 and adding an isotonic regulator of appropri...

Embodiment 5

[0043] Screening test and comparative verification experiment of embodiment 5 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 , record the key factor K value. We take K=1000 (combining the technical requirements of many different manufacturers) as the judgment standard, if it exceeds this value, it means that the reagent does not meet the requirements. Only list the comparative effect of 2 groups of comparative examples below, but will know that concrete comparative example also comprises the combination of many other multiple different stabilizers, and wherein the formula of embodiment and comparative example is shown in Table 4, in the process of accelerated destruction experiment, K value See Table 5 for the changes.

[0045] Table 4

[0...

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Abstract

The invention relates to the technical field of detection of in vitro diagnostic reagents, and particularly relates to a total bile acid detection kit. The total bile acid detection kit comprises a reagent R1 and a reagent R2, wherein the reagent R1 is prepared from a buffer solution, a surfactant, a preservative, an isoosmotic adjusting agent and a Thio-NAD; the reagent A2 is prepared from the buffer solution, the surfactant, a stabilizing agent, the preservative, reduced coenzyme I and 3 alpha-hydroxysteroid dehydrogenase; the buffer solution in the reagent R1 is a glycine buffer solution, acitric acid-sodium citrate buffer solution, a potassium hydrogen phthalate-sodium hydroxide buffer solution, a PBS (Phosphate Buffered Saline) buffer solution and the like; the isoosmotic adjusting agent in the reagent R1 is one or more of NaCl, KC1 and MgCl2; the stabilizing agent in the reagent R2 is TEA, BSA, TCEP, glutathione, mannitol and the like. When the total bile acid of a special sample is tested by utilizing the kit disclosed by the invention, a reaction curve can be improved, and the kit can be placed for a long time, so that the 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 acids are an important component of bile and play an important role in fat metabolism. Bile acids mainly exist in the enterohepatic circulatory system and play a certain protective role through recirculation. Only a small part of bile acids enters the peripheral circulation and promotes enterohepatic circulation. The hepatic circulation is powered by the hepatocyte transport system, uptake of bile acids and secretion into bile, cholecystokinin-induced gallbladder contraction, propulsive peristalsis of the small intestine, active transport of the ileal mucosa, and influx 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) and deoxycholic acid (DCA) produced in metabolism, a...

Claims

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Application Information

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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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