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Kit and method for detecting concentration of fibronectin in urine

A technology of fibronectin and kit, which is applied in the field of bioengineering, can solve the problems of complex concentration process and low accuracy, and achieve the effect of simple and convenient process, high accuracy and remarkable technological progress

Inactive Publication Date: 2014-03-26
上海北加生化试剂有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] In view of the above-mentioned defects in the prior art, the technical problem to be solved by the present invention is to provide a kit for detecting the concentration of fibronectin in urine and a method thereof. The kit for connexin concentration and its method should solve the technical problems of complex process and low accuracy in the determination of urinary fibronectin (FN) concentration by immunodiffusion method and ELISA double-antibody sandwich technology in the prior art

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  • Kit and method for detecting concentration of fibronectin in urine
  • Kit and method for detecting concentration of fibronectin in urine
  • Kit and method for detecting concentration of fibronectin in urine

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Experimental program
Comparison scheme
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Embodiment 1

[0021] The invention provides a kit for detecting the concentration of fibronectin in urine, comprising two reagents, reagent I consists of disodium hydrogen phosphate, potassium dihydrogen phosphate, PEG6000-8000, disodium edetate ( EDTA-NA 2 ) and Triton 100 (TX-100), in the reagent I, the mass percentage concentration of disodium hydrogen phosphate is 28.6g / L, and the mass percentage concentration of potassium dihydrogen phosphate is 2.7g / L, the mass percentage concentration of described PEG6000 is 50g / L, described EDTA-NA 2 The mass percentage concentration of TX-100 is 1g / L, the volume concentration of the TX-100 is 0.2ml / L, and the II reagent is rabbit anti-human fibronectin antiserum, or goat anti-human fibronectin antiserum, or mouse anti-human Suspension of fibronectin antiserum, or mouse anti-human fibronectin monoclonal antibody cross-linked on latex particles.

[0022] Further, the II reagent is a rabbit anti-human fibronectin antiserum, or goat anti-human fibron...

Embodiment 2

[0027] 2.1 Applicable instruments

[0028] Semi-automatic and fully automatic biochemical analyzers.

[0029] 2.2 Analysis method

[0030] Immunoturbidimetry.

[0031] 2.3 Performance requirements

[0032] 2.3.1 Reagent Appearance

[0033] R1: colorless clear transparent liquid;

[0034] R2: milky white suspension.

[0035] 2.3.2 Reagent blank absorbance (A)

[0036] Absorbance (A): R 1 +R 2 ≤2600A (at a temperature of 37°C; main wavelength 546nm, 600nm (secondary wavelength 700nm)).

[0037] 2.3.3 Precision

[0038] 2.3.3.1 Intra-assay precision

[0039] CV≤10%.

[0040] 2.3.4 Batch-to-batch precision

[0041] The relative range is ≤10%.

[0042] 2.3.5 Accuracy

[0043] Inaccuracy: within the range of ±10%.

[0044] 2.3.6 Analytical sensitivity

[0045] Absorbance (A)>2600A.

[0046] 2.3.7 Linearity

[0047] In the range of 20mg / L-100mg / L, the correlation coefficient (γ)≥0.9900.

[0048] 2.3.8 Stability

[0049] The reagent should be stored in the dark at ...

Embodiment 3

[0050] Embodiment 3 Experimental method

[0051] 3.1 Detection conditions

[0052] 3.1.1 Biochemical analyzer (hereinafter referred to as instrument)

[0053] Hitachi 7060 automatic biochemical analyzer.

[0054] 3.1.2 Working environment temperature

[0055] Room temperature 15-32°C.

[0056] Indoor humidity 45-85%RH.

[0057] 3.1.3 Main measurement parameters and operation steps

[0058]Temperature 37°C; main wavelength 600nm (secondary wavelength 700nm), urine sample or urine calibration standard 3μl; R 1 : 240 μl; R 2 : 60 μl, reaction time 10 minutes.

[0059] Method type: two-point endpoint method.

[0060] After adding reagent R1 to the urine sample or urine calibrator, incubate at 37°C for 5 minutes, read the first point (A1), add reagent R2, incubate at 37°C for 5 minutes, and read the second point (A2), sample A=A2-A1.

[0061] Result calculation:

[0062]

[0063] 3.1.4 Calibrator

[0064] The calibrator used in this standard is the standard product pr...

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Abstract

The invention provides a kit for detecting the concentration of fibronectin in urine, which comprises two reagents. A reagent I comprises phosphate buffer; a reagent II is suspension formed by cross-linking a rabbit anti-human fibronectin antiserum, a goat anti-human fibronectin antiserum, a rat anti-human fibronectin antiserum or a rat anti-human fibronectin monoclonal antibody on latex particles. The invention solves the problems of complex steps, low accuracy and poor repeatability when a radial immunodiffusion method and an ELISA (Enzyme Linked Immunosorbent Assay) double-antibody sandwich technology are adopted to measure the concentration of the fibronectin in the prior art. The invention also provides a method for detecting the concentration of the fibronectin in human urine by adopting the kit. The kit and the method which are provided by the invention are used for measuring the concentration of the fibronectin in the urine, adopt simple steps, have high accuracy and good repeatability and are used for an automatic analysis meter.

Description

technical field [0001] The invention belongs to the field of bioengineering, and in particular relates to a detection kit, in particular to a kit and a method for detecting the concentration of fibronectin (FN) in human urine by using enhanced latex immunoturbidimetry. Background technique [0002] Fibronectin, also known as fibronectin (FN for short), consists of two similar polypeptide chain subunits, and the molecular weight of the subunits is 220-250KD. In the prior art, the immunodiffusion method and ELISA double-antibody sandwich technology are used to measure the concentration of fibronectin (FN) in urine. The disadvantages are cumbersome operation, high work intensity, and long retention time. Generally, it takes 8-48 hours. The accuracy is poor. [0003] Because of the low concentration of fibronectin (FN) in urine, there have been no reports on the use of this kit and method at home and abroad. Contents of the invention [0004] In view of the above-mentioned d...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N21/31
CPCG01N33/6893G01N33/54313G01N33/57407
Inventor 杨杰王泉龙朱晓敏刘颖冰
Owner 上海北加生化试剂有限公司
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