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Kit and method for detecting C1q concentration of complement with nanosized-latex enhancing immune turbidimetry

An immunoturbidimetric and nano-latex technology, applied in the field of bioengineering, can solve the problems of complex Clq concentration process and low accuracy of complement, and achieve the effects of high accuracy and simple and convenient process.

Active Publication Date: 2014-06-18
上海北加生化试剂有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The object of the present invention is to provide a kind of test kit and the method that strengthen nano-latex immune turbidimetry to detect the concentration of complement Clq, and the described kit and the method of this enhanced nano-latex immune turbidimetry to detect the concentration of complement Clq will solve existing In the technology, the immunodiffusion method and ELISA double antibody sandwich technology are used to measure the concentration of complement Clq, which is complicated and has low accuracy.

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  • Kit and method for detecting C1q concentration of complement with nanosized-latex enhancing immune turbidimetry
  • Kit and method for detecting C1q concentration of complement with nanosized-latex enhancing immune turbidimetry
  • Kit and method for detecting C1q concentration of complement with nanosized-latex enhancing immune turbidimetry

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] 1.1 Applicable instruments

[0019] Semi-automatic and fully automatic biochemical analyzers.

[0020] 1.2 Analysis method

[0021] Enhanced Nanolatex Immunoturbidimetry.

[0022] 1.3 Performance requirements

[0023] 1.3.1 Reagent Appearance

[0024] R1: colorless clear transparent liquid;

[0025] R2: Pale yellow clear liquid or latex suspension.

[0026] 1.3.2 Reagent blank absorbance (A)

[0027] Absorbance (A): R 1 +R 2 ≤0.04A (at a temperature of 37°C; main wavelength 546nm, 600nm (secondary wavelength 700nm)).

[0028] 1.3.3 Precision

[0029] 1.3.3.1 Intra-assay precision

[0030] CV≤10%.

[0031] 1.3.4 Batch-to-batch precision

[0032] Relative range ≤ 10%.

[0033] 1.3.5 Accuracy

[0034] Inaccuracy: within the range of ±10%.

[0035] 1.3.6 Analytical sensitivity

[0036] Absorbance (A)>0.04A.

[0037] 1.3.7 Linearity

[0038] In the range of 50mg / L-400mg / L, the correlation coefficient (γ)≥0.9900.

[0039] 1.3.8 Stability

[0040] The reage...

Embodiment 2

[0043] 2.1 Detection conditions

[0044] 2.1.1 Biochemical analyzer (hereinafter referred to as instrument)

[0045] Hitachi 7060 automatic biochemical analyzer.

[0046] 2.1.2 Working environment temperature

[0047] Room temperature 15-32°C.

[0048] Indoor humidity 45-85% RH.

[0049] 2.1.3 Main measurement parameters and operation steps

[0050] Temperature 37°C; main wavelength 546nm, 600nm (secondary wavelength 700nm), sample or calibrator 3-4μl; R 1 : 240 μl; R 2 : 60 μl, reaction time 10 minutes.

[0051] Method type: two-point endpoint method.

[0052] After adding reagent R1 to the sample or 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.

[0053] Result calculation:

[0054]

[0055] 2.1.4 Calibrator

[0056] The calibrator used in this standard is the standard product produced by Nanfang Reagent Factory in Yuhuan County, Zhejiang Provi...

Embodiment 3

[0096] Preparation of reagents

[0097] Take a sample with a content of about 200mg / L, which is sample 4 # , the sample 4 # Prepare 5 test samples with physiological saline or pure water according to the method in the table below. (This sample is ready for use now)

[0098]

[0099] After adding reagent R1 to the sample or 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.

[0100] Result instrument calculation:

[0101]

[0102] Through the determination of the above samples, the figure 1 Dose-response curves of complement Clq concentration and absorbance are shown.

[0103] ( figure 1 The curve is based on the OD value of the wavelength of 600nm, and the sub-wavelength of 700nm is for reference only. )

[0104]

[0105] pass figure 1 , measured the serum of 500 cases of normal people, the results are described in the following table, ac...

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Abstract

The invention belongs to the field of biological engineering, and provides a kit for detecting C1q concentration of a complement in blood serum with a nanosized-latex enhancing immune turbidimetry, solving the problems of complex steps, low accuracy and poor repeatability in the prior art adopting an immunodiffusion method and an ELISA (Enzyme-Linked Immuno Sorbent Assay) double-antibody sandwich technology to detect the C1q concentration of the complement. The kit comprises two reagents, wherein a reagent I is prepared from disodium hydrogen phosphate, monopotassium phosphate, PEG6000, EDTA-NA2 and TX-100; and a reagent II is prepared from suspensions of rabbit anti-human complement C1q antiserum or goat anti-human complement C1q antiserum or rat anti-human complement C1q antiserum or rat anti-human complement C1q monoclonal antibodies crosslinking on latex particles. The invention also provides a method using the kit to detect the C1q concentration of the complement in human blood serum. The kit and the method have simpleness and convenience in steps for detecting the C1q concentration of the complement, high degree of accuracy and good repeatability and are used for automatic analyzers.

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 complement Clq in human serum and joint cavity fluid by using enhanced nano-latex immunoturbidimetry. Background technique [0002] Complement Clq is the first component of Cl in the complement system. It is a glycoprotein with a large molecular weight. One Clq molecule is composed of 18 polypeptide chains. The chemical composition is collagen protein molecular weight: 410KD. In the prior art, immunodiffusion method and ELISA double-antibody sandwich technology are used to measure the concentration of complement Clq, but the steps are complicated and the accuracy is not high. Contents of the invention [0003] The object of the present invention is to provide a kind of test kit and the method that strengthen nano-latex immune turbidimetry to detect the concentration of complement Cl...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68
Inventor 杨杰朱晓敏朱慧琳张瑞镐王泉龙刘颖冰
Owner 上海北加生化试剂有限公司
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