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Latex-enhanced immunonephelometry kit for determination of resistin, preparation method and detection method thereof

An immunoturbidimetric and latex-enhanced technology, which is applied in the field of disease diagnosis or detection, can solve the problems such as the absence of resistin kits, and achieve the effect of improving accuracy

Inactive Publication Date: 2015-08-12
NANJING WENZHI BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] At present, there is no detection method and corresponding kit that can quantitatively measure resistin in the domestic market

Method used

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  • Latex-enhanced immunonephelometry kit for determination of resistin, preparation method and detection method thereof
  • Latex-enhanced immunonephelometry kit for determination of resistin, preparation method and detection method thereof
  • Latex-enhanced immunonephelometry kit for determination of resistin, preparation method and detection method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1 Preparation of a latex-enhanced immune turbidimetric kit for the determination of resistin

[0029] 1. Reagent configuration

[0030] 1.1. Preparation of diluent R1

[0031] Recipe: 0.05M Tris-HCl buffer containing 2M NaCl, 2% (w / w) PEG20000, 5% (w / w) BSA, 0.1% (w / w) proclin-300, pH=8.4.

[0032] 1.2. Preparation of latex reagent R2

[0033] (1) Carboxylated polystyrene latex activation

[0034] Add 50mM in the carboxylated polystyrene latex 1ml of 10mg / ml, 1mg EDC (1-(3-dimethylaminopropyl group)-3-ethylcarbodiimide hydrochloride) and 10mg that PH6.1MES dissolves NHS (N-hydroxysuccinimide) was mixed and activated for 60 minutes, centrifuged at 19750rpm to discard the supernatant, and resuspended with 50mM PBS; according to the above method, the activated carboxylated The polystyrene latex, the particle diameter of the latex used in this experiment is respectively 60nm and 150nm;

[0035] (2) Preparation of carboxylated polystyrene latex coupled with resi...

Embodiment 2 Embodiment 1

[0066] Embodiment 2 Embodiment one kit stability test

[0067] The storage condition of the kit is 2-8°C. After 6 months of storage, the various indicators of the kit were measured and found to be within the normal range. Considering that there will be abnormal storage conditions during transportation and use, the kit was stored at 37°C for 6 days, and the accelerated aging test was carried out. The results showed that the indicators of the kit fully met the requirements. Considering the freezing of the kit, the kit was put into a -20°C refrigerator for 5 days, and the test results also showed that all the indicators of the kit were completely normal. From the above results, it can be concluded that the kit can be stored at 2-8°C for at least 6 months.

Embodiment 3

[0068] Embodiment 3 The detection method of the kit of Embodiment 1 of the present invention

[0069] The immunoturbidimetric method is adopted, and the latex-enhanced immune turbidimetric method is used as the determination principle. The resistin in the sample reacts specifically with the latex particles labeled with resistin antibody to form an immune complex, which causes a change in absorbance, which is proportional to the concentration of resistin in the sample. Use the resistin standard substance of known concentration to make working curve, can calculate the resistin content in the sample from this curve, the standard curve prepared by the calibrator of the present invention is as follows figure 1 As shown, after the calibration is completed, first test the quality control product, and its concentration is within the range marked (0.20-0.30μg / L), and the kit is valid.

[0070] 1. Applicable instruments: Toshiba (TBA-40FR), Dirui (CS-T300), Laura (FAITH-1000), Olympus ...

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Abstract

The invention discloses a latex-enhanced immunonephelometry kit for the determination of resistin, a preparation method and a detection method thereof, and belongs to the technical field of disease diagnosis and detection. The latex-enhanced immunonephelometry kit for the determination of resistin comprises a diluent, a latex preparation, a blank solution, a calibrator and a quality control; the latex preparation contains carboxylated polystyrene latex with different particle sizes and respectively coupled with resistin monoclonal antibodies and resistin polyclonal antibodies. After labeling of the resistin monoclonal antibodies and resistin polyclonal antibodies, the obtained latex with large particle size can improve detection sensitivity, and the latex with small particle size can broaden the linear range; therefore, the composite latex preparation can enhance the detection sensitivity, also broaden the linear range of detection, and has the advantages of rapidness, high sensitivity, strong specificity and good accuracy for resistin detection.

Description

technical field [0001] The invention discloses a latex-enhanced immune turbidimetric kit and its preparation method and detection method, in particular to a latex-enhanced immune turbidimetric kit for measuring resistin and its preparation method and detection method. The invention belongs to disease in the field of diagnosis or detection technology. Background technique [0002] Resistin (resistin) is a new cytokine found in mouse adipocytes by Steppan et al. in 2001. Because it has the effect of reducing insulin sensitivity, it is named resistin. It belongs to the secreted protein family of resistin-like molecules (resistin-like molecules, RELMs). [0003] Human resistin is a 12.5kDa protein, and its precursor contains 108 amino acids. The resistin secreted into the blood has two states, namely hexamer and trimer. It participates in the inflammatory response, interacts with some cytokines, inflammatory mediators, and adhesion molecules, and can promote the synthesis and ...

Claims

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Application Information

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IPC IPC(8): G01N33/68
CPCG01N33/6893G01N2333/715
Inventor 许佩佩
Owner NANJING WENZHI BIOTECH
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