Agent kit for measuring urinary transferrin by latex-enhanced immunoturbidimetry and preparation method thereof

A technology of transferrin and immunoturbidimetry, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., and can solve the problems of no routine detection of urine transferrin

Inactive Publication Date: 2011-07-20
上海景源医疗器械有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In order to solve the above problems, the present invention improves the sensitivity of detection by studying the method of latex particle enhancement, thereby solving the problem that there is no means for routine detection of urine transferrin in clinical practice, and provides a latex-enhanced immune turbidimetric method for the determination of urine transferrin. Ferritin Kit

Method used

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  • Agent kit for measuring urinary transferrin by latex-enhanced immunoturbidimetry and preparation method thereof
  • Agent kit for measuring urinary transferrin by latex-enhanced immunoturbidimetry and preparation method thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0032] (1) Anti-transferrin antibody coated latex particles

[0033] A latex-enhanced immune turbidimetric assay urine transferrin kit is characterized in that the kit includes latex particles, transferrin antibodies and a buffer; transferrin antibodies and latex particles are covalently bonded The approach is to establish a low-concentration transferrin level and a transferrin antibody that has been bound to latex particles to react to form a 340nm-700nm detectable turbidity reaction system, including the following steps:

[0034] 1. Activation of latex particles

[0035] (1) Add 1000 μl of Q0327 (rabbit anti-human transferrin antibody, produced by Dako, Denmark) to 2000 μl of 20 mM phosphate buffer, pH 7.50 (code 20PS5), mix well and set aside;

[0036] (2) Take 600 μl of 10% K1-0005 latex particles (manufactured by Merck), centrifuge at 21,000 rpm for 90 min at 4°C, and discard the filtrate;

[0037] (3) Add 1000 μl of 10mM phosphate buffer solution, pH 6.00 (code 10PS), ...

Embodiment 2

[0059] Example 2: Urinary Transferrin Immunoturbidimetric Detection Kit Using Steps and Detection of Analytical Performance

[0060] 1 detection conditions

[0061] 1.1 Instrument

[0062] Olympus automatic biochemical analyzer

[0063] 1.2 Detection settings

[0064] Wavelength: 340nm

[0065] Detection dosage: 30+150+50

[0066] Reaction buffer: 200mmol / L, Tris solution pH7.15 (code RB)

[0067] 2. Detection

[0068] 2.1 Determination method

[0069] a) Mark test tubes: standard tubes, sample tubes, quality control tubes.

[0070] b) Zero the spectrophotometer at 340 nm with distilled water.

[0071] c) Add 30 μL pre-diluted standard solution, sample and quality control solution to the corresponding test tubes respectively.

[0072] d) Add 150 μL transferrin reagent 1 to each tube, mix well, pre-warm at 37°C for 3-5 minutes, and measure the absorbance A1.

[0073] e) Add 30 μL of transferrin reagent 2 to each tube, mix well and react at 37° C. for 5 minutes, and meas...

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Abstract

The invention provides an agent kit for measuring urinary transferrin by latex-enhanced immunoturbidimetry and a preparation method thereof. The agent kit is characterized by comprising emulsion particles, transferrin antibody and buffer liquid; according to appropriate paths for covalent bond between the transferrin antibody and the emulsion particles, a turbidity reaction system between 340-700 nm detection is established below the low-concentration transferrin level and the transferrin antibody bonded with the emulsion particles. The preparation method comprises the following steps of: activation of emulsion particles; bonding of the transferrin antibody and the emulsion particles; and storing. The agent kit can be used for detecting the urinary transferrin level with the concentration to be 0-12 mg / L in the urine; the accuracy can trace back to CRM470, and the precision is smaller than 10% so as to satisfy the clinic requirements.

Description

technical field [0001] The invention relates to a latex-enhanced immune turbidimetric assay kit for urine transferrin, in particular to a latex particle enhancement technology to improve the sensitivity of transferrin concentration detection, which can be used for the detection of low-concentration transferrin in urine . Background technique [0002] The glomerular filtration membrane has two barriers for protein filtration, namely volume barrier and charge barrier. The size of the filtration pore size of the filtration membrane determines the volume barrier; the charge barrier arises because the glomerular filtration membrane is rich in negatively charged structural components, making it difficult for negatively charged proteins to filter, transferrin belongs to this category protein. It has been proved that once the glomerulus is diseased, the content of negatively charged components sialic acid and heparan sulfate in the glomerular filtration membrane decreases, and the...

Claims

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Application Information

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IPC IPC(8): G01N33/545
Inventor 孙永杰
Owner 上海景源医疗器械有限公司
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