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Serum lipase detection kit as well as preparation method and application thereof

A technology for detection kits and detection methods, applied in biochemical equipment and methods, measuring devices, microbial determination/inspection, etc., can solve the problems of increased reagent consumption, loss of manpower and material resources, poor stability of determination methods, Solve problems such as batch-to-batch variability of chromophore substrates, reduce reagent consumption, manpower and material resources, avoid repeated testing, and achieve good reproducibility

Pending Publication Date: 2020-03-27
ZHONGSHAN CHUANGYI BIOCHEM ENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Most of the commercial lipase detection kits on the market have a narrow detection range (only 5-300U / L), and the more high-value samples (that is, above 300U / L) appear in the clinical detection process, resulting in its daily During the detection process, the frequency of repeated detection is increasing day by day, which increases the consumption of reagents, manpower and material resources.
In addition, batch-to-batch variability still exists in the emulsification of chromophore substrates, which leads to poor stability of the assay method, and these defects still limit its application range
The reasons may be: 1) in order to ensure high sensitivity, the upper limit of absorbance exceeds the limit when measuring high-value samples; 2) more lipase substrates need to be dissolved in the reagent to measure high-value samples, and the substrate Increases can easily lead to substrate precipitation and cause kit failure
[0008] Chinese invention patent CN104215632A discloses a stable lipase kit, which solves the reagent stability, but its quantitative range is still narrow

Method used

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  • Serum lipase detection kit as well as preparation method and application thereof
  • Serum lipase detection kit as well as preparation method and application thereof
  • Serum lipase detection kit as well as preparation method and application thereof

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Experimental program
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Effect test

Embodiment 1

[0043] Embodiment 1 of the present invention is: a serum lipase detection kit and its preparation method and application, the kit is composed of reagent R1, reagent R2 and calibrator. The calibrator was a biochemical compound calibrator produced by Landau Company in the United Kingdom. The composition of reagent R1 is as follows:

[0044] pH 8.8N,N-bis(2-hydroxyethyl)-glycine buffer 50mmol / L taurodeoxycholic acid 25mmol / L Sodium deoxycholate 5mmol / L Calcium Chloride Anhydrous 5mmol / L Proclin300 0.05％

[0045] The composition of reagent R2 is as follows:

[0046] pH 5.5 tartrate buffer 10mmol / L 1,2-Dilaurylglycerol-3-glutaric acid-(6’-methylresorufin)-ester 0.25mmol / L Mannitol 1％ Triton-100 0.05％ DMSO 1.0％ n-propanol 0.5％ Lecithin 0.2％ BSA 1％

[0047] The preparation process of the kit is as follows:

[0048] Reagent R1 can be prepared according to the conventional mixing se...

Embodiment 2

[0056] Embodiment two of the present invention is: the accuracy test of the kit that above-mentioned embodiment 1 makes:

[0057] Since the lipase item cannot be traced to the SI unit, the accuracy test was carried out through a comparative test.

[0058] 1. After using the calibrator to calibrate, conduct a comparison experiment with the same batch of serum measured by a mature kit on the market. The results are shown in Table 2 below:

[0059] Table 2

[0060]

[0061]

[0062] Mark the data points in the above table in the two-dimensional plane coordinate system, the result is as follows figure 2 Shown (in the figure x represents the concentration measured by the mature kit in the market, and the y value represents the concentration measured by the program kit of the present invention), each point is carried out by linear correlation fitting and found that its slope is close to 1, and the intercept is also It is close to zero, which shows that the detection result ...

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Abstract

The invention discloses a serum lipase detection kit as well as a preparation method and application thereof. The kit comprises a reagent R1 and a reagent R2, wherein the reagent R2 comprises the following components: a tartaric acid buffer solution of which the pH value is 5.5, 1,2-dilauryl glycerinum-3-glutaric acid-(6'-methyl resorufin)-ester, mannitol, triton-100, DMSO (dimethyl sulfoxide), normal propyl alcohol, lecithin and BSA (bull serum albumin). The kit is wide in linear range (100-700U / L), low in cost, good in stability and high in sensitivity, and has good market popularization application prospects.

Description

technical field [0001] The invention relates to the technical field of in vitro diagnosis, in particular to a serum lipase detection kit and its preparation method and application. Background technique [0002] Lipase belongs to the carboxyl ester hydrolase class, which can gradually hydrolyze triglycerides into glycerol and fatty acids. Lipase exists in fat-containing animal, plant and microbial (such as mold, bacteria, etc.) tissues. Lipase (Lipase, LPS) is a group of lipolytic enzymes with low specificity, mainly derived from the pancreas, followed by the stomach and small intestine, and can hydrolyze a variety of glycerides containing long-chain (8-18 carbon chain) fatty acids. LPS needs to be distinguished from another esterase with very low specificity. Esterase acts on esters containing short-chain fatty acids that are soluble in water; lipase only acts on fat at the interface between ester and water, and LPS only works when the substrate is in an emulsion state. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/44
CPCC12Q1/44G01N2333/92
Inventor 于云飞崔海林曾晓君黄清媚
Owner ZHONGSHAN CHUANGYI BIOCHEM ENG
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