Fluorescence method and kit for testing lymphocyte homing receptor

A technology for homing receptors and lymphocytes, applied in fluorescence/phosphorescence, material excitation analysis, etc., can solve the problems of time-consuming operation and high cost, and achieve the effect of simple operation, low cost and fast response speed.

Inactive Publication Date: 2014-03-26
深圳云安智慧医疗科技有限公司
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

Although this method can achieve high sensitivity and excellent specificity, it is time-consuming and costly

Method used

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  • Fluorescence method and kit for testing lymphocyte homing receptor
  • Fluorescence method and kit for testing lymphocyte homing receptor
  • Fluorescence method and kit for testing lymphocyte homing receptor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Example 1 Detection of different concentrations of CD44

[0044] In the Tris-HCl buffer solution with a temperature of 25°C and pH=8, add 5×10 -7 mol / L cationic water-soluble conjugated polymer and 2.5×10 -5 A series of different concentrations of CD44 were added to the bottom solution of the mol / L hyaluronic acid probe, the concentrations were 1.95×10 -7 , 3.9×10 -7 , 6.5×10 -7 , 1.0×10 -6 , 1.0×10 -5 g / mL, scan its fluorescence emission spectrum, the experimental results are as follows Figure 5 shown. It can be seen from the figure that as the concentration of CD44 increases, the FRET efficiency from the cationic water-soluble conjugated polymer to the hyaluronic acid probe decreases gradually. ΔFRET was calculated and plotted against the concentration of CD44 to obtain Figure 6 . The figure shows that at a CD44 concentration of 10 -7 There is a very good linear range in the range of g / mL order of magnitude, and the curve of ΔFRET tends to be flat when it ...

Embodiment 2

[0046] Embodiment 2 specificity analysis

[0047] Detection 1: Set the concentration to 1.95×10 -7 g / mL of CD44 was added to the detection bottom solution, and other experimental conditions were the same as in Example 1.

[0048] At the same time, do the following test as a comparison.

[0049] Detection 2: An equal volume of bovine serum albumin (BSA) and thrombin (Thrombin) with a concentration 100 times higher than that of CD44 was used for the test as a negative control, and other experimental conditions were the same as in Example 1.

[0050] Detection 3: An equal volume of blank buffer solution was used instead of CD44 for the test as a blank control, and other experimental conditions were the same as in Example 1.

[0051] The result is as Figure 7 shown. The relative change of fluorescence resonance energy transfer efficiency of the detection system added with CD44 was significantly higher than that of the other three control systems, indicating that this method h...

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Abstract

The invention discloses a fluorescence method a kit for testing a lymphocyte homing receptor, the fluorescence method is as follows: a cationic conjugated polymer is bonded with fluorescein-marked negatively-charged hyaluronic acid by electrostatic attraction for fluorescence resonance energy transfer from the conjugated polymer to the fluorescein, and the fluorescence intensity of the fluorescein is increased significantly; when CD44 ( lymphocyte homing receptor) is added, specific bonding of the CD44 and the hyaluronic acid is happened, the bonding of the cationic conjugated polymer and the hyaluronic acid with fluorescein groups is weakened, the distance between molecules is increased, so that the energy transfer efficiency is reduced, the conjugated polymer fluorescence is gradually restored, the fluorescein fluorescence is gradually weakened, and the energy transfer efficiency reducing degree is relative to the concentration of the CD44. The method is simple in operation, fast in response speed, low in cost and high in sensitivity and selectivity, and has important significances in biological detection and early diagnosis and treatment of diseases.

Description

technical field [0001] The invention belongs to the field of biosensing and analysis, and in particular relates to a fluorescent method and a kit for detecting lymphocyte homing receptor (CD44) by using a cationic conjugated polymer and a hyaluronic acid probe. Background technique [0002] The light-absorbing ability of conjugated polymers is very strong, and it also has the function of "molecular wires", which allows the collected light energy or photo-excited electrons to migrate and transfer rapidly on the conjugated main chain, thereby rapidly generating group responses to the detected molecules, reflecting the Unique fluorescent signal amplification. Therefore, conjugated polymers have become popular reporter materials in the field of sensing in recent years. Compared with traditional small molecule fluorescent materials, they often have better signal multiplication effect and real-time detection effect. In order to apply conjugated polymers to biosensing, ionic group...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64
Inventor 黄艳琴欧阳浪黄维刘兴奋范曲立
Owner 深圳云安智慧医疗科技有限公司
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