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A stable serum phospholipid detecting reagent high in interference-resisting capability and a detecting method

A technology for serum phospholipids and detection reagents, which is applied in biochemical equipment and methods, microbiological determination/inspection, etc. It can solve the problems of reagents that are harmful to operators, difficult to realize automatic operation, poor stability of reagents, etc., and achieve enhanced anti-interference Ability, improve anti-interference ability, improve the effect of stability

Active Publication Date: 2016-05-04
BIOBASE BIODUSTRY (SHANDONG) CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Chemical method, the method is stable, but the operation is cumbersome, the reagents are more harmful to the operator, easy to cause environmental pollution, and it is difficult to realize automatic operation
The enzymatic reagents are easy to operate, high in sensitivity, and suitable for automatic analysis, but the stability of the reagents is slightly poor, and they are easily interfered by interfering substances such as bilirubin and ascorbic acid.

Method used

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  • A stable serum phospholipid detecting reagent high in interference-resisting capability and a detecting method
  • A stable serum phospholipid detecting reagent high in interference-resisting capability and a detecting method
  • A stable serum phospholipid detecting reagent high in interference-resisting capability and a detecting method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] Serum phospholipid detection reagents, including reagent R1 and reagent R2:

[0061] 1) The composition of its R1 is:

[0062] HEPES (4-hydroxyethylpiperazineethanesulfonic acid) buffer solution (pH=7.2, 25°C) · 100mmol / L,

[0063] Phospholipase D ······································· 1KU / L,

[0064] DAOS ·········································· 1.4mmol / L,

[0065] ascorbate oxidase ································ 2KU / L,

[0066] Bilirubin oxidase ·································· 2KU / L,

[0067] polyethylene glycol 6000 ·································· 10g / L,

[0068] sucrose ·········································· 20g / L,

[0069] xanthan gum ········································ 0.5g / L,

[0070] Mannitol ········································ 20g / L,

[0071] Trehalose ········································ 10g / L,

[0072] BSA ··········································· 2g / L,

[0073] Polyoxypropylene oxide ethylene glyceryl ether (P...

Embodiment 2

[0094] Interference test: take fresh mixed serum, divide it into 2 equal parts, then divide each equal part into 5 equal parts, add different interfering substances, so that the concentration in the serum reaches the requirements in Table 2. Then, the reagents obtained in Example 1 were used to compare and measure the content of PLIP in serum at the same time as the common and recognized serum phospholipid (PLIP) reagent in the market. Relative deviation (%) = (measuring mean value of interference samples - measuring mean value of control samples) / measured mean value of control samples × 100%.

[0095] It can be seen from Table 2 that the reagent of Example 1 has no obvious interference on the test results when ascorbic acid≤50mg / dL, bilirubin≤40mg / dL, triglyceride≤1250mg / dL, and hemoglobin≤400mg / dL. However, the reagents of the control group were significantly disturbed when the above-mentioned concentration of interfering substances existed, which indicated that by optimizin...

Embodiment 3

[0099] Correlation experiment: using the formula in Example 1 to prepare reagents, and conducting a control test with a phospholipid kit from a company approved by the State Food and Drug Administration, which is common in the market, and testing 20 clinical serum samples at the same time, the test results are shown in Table 3 . 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 0.9997, indicating that there is a great correlation between the two.

[0100] Table 3 Example 1 reagents and market common and recognized serum phospholipid assay kit comparative detection results

[0101]

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Abstract

The invention relates to the technical field of serum phospholipid detection and particularly relates to a serum phospholipid detecting reagent. A reagent R1 comprises a buffer solution, phospholipase D, DAOS, ascorbic acid oxidase, bilirubin oxidase, polyethylene glycol 6000, cane sugar, xanthan gum, mannitol, trehalose, BSA, glycerol propoxylate-block-ethoxylate (Pluranic L64) and an aseptic. A reagent R2 comprises the buffer solution, 4-aminoantipyrine, peroxidase, choline oxidase, the polyethylene glycol 6000, the cane sugar, the xanthan gum, the mannitol, the trehalose, the BSA, the glycerol propoxylate-block-ethoxylate (Pluranic L64) and the aseptic. The HEPES buffer solutions and a novel Trinder reaction chromogen DAOS are adopted. A plurality of stabilizers are added to obviously improve stability of the detecting reagent. The bilirubin oxidase and the ascorbic acid oxidase are added, thus effectively avoiding interference caused by bilirubin and ascorbic acid and greatly improving interference-resisting capability of the detecting reagent. In addition, addition of the glycerol propoxylate-block-ethoxylate (Pluranic L64) which is an optional nonionic surfactant prevents reaction system turbidity, enhances substrate stability and improves the interference-resisting capability of the detecting reagent.

Description

technical field [0001] The invention relates to the technical field of serum phospholipid detection, in particular to a serum phospholipid detection reagent and a detection method using the detection reagent. Background technique [0002] Serum phospholipids mainly include four parts: lecithin, lysolecithin, neurophospholipid and cephalin. In clinical work, only serum total phospholipids are generally measured. The increase of serum phospholipids is seen in diabetes, nephrotic syndrome, chronic hemorrhagic anemia, liver cirrhosis, liver necrosis, biliary obstruction, hypothyroidism, essential hypertension, syphilis, etc.; Hyperthyroidism, acute infectious fever, malnourished liver cirrhosis, etc. [0003] At present, there are chemical methods and enzymatic methods for the detection of serum total phospholipids, and now the enzymatic method is the most commonly used. Chemical method, the method is stable, but the operation is cumbersome, the reagents are more harmful to th...

Claims

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Application Information

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IPC IPC(8): C12Q1/44C12Q1/28C12Q1/26
CPCC12Q1/26C12Q1/28C12Q1/44C12Q2326/96
Inventor 李志明甘宜梧赵新李静
Owner BIOBASE BIODUSTRY (SHANDONG) CO LTD
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