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Kit and method for detecting concentration of complement Clq in human serum

A technology for detecting human and concentration, which is applied in the field of bioengineering, can solve the problems of complicated process and low accuracy of complement Clq concentration, and achieve the effect of significant technological progress, high accuracy, and simple and convenient process

Active Publication Date: 2013-05-22
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

[0003] The purpose of the present invention is to provide a test kit for detecting the concentration of complement Clq in human serum and a method thereof. The test kit and method for detecting the concentration of complement Clq in human serum should solve the problem of using the immunodiffusion method in the prior art. The process of determining the concentration of complement Clq with ELISA double-antibody sandwich technology is complicated and the accuracy is not high.

Method used

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  • Kit and method for detecting concentration of complement Clq in human serum
  • Kit and method for detecting concentration of complement Clq in human serum
  • Kit and method for detecting concentration of complement Clq in human serum

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] 1.1 Kit specifications

[0017]

[0018] 1.2 Components

[0019] R 1 : Disodium hydrogen phosphate 28.6g / L Potassium dihydrogen phosphate 2.7g / L

[0020] PEG6000 50g / L EDTA-NA 2 1g / L

[0021] TX-100 0.2ml / L

[0022] R 2 : Disodium hydrogen phosphate 28.6g / L Potassium dihydrogen phosphate 2.7g / L

[0023] PEG6000 50g / L EDTA-NA 2 1g / L

[0024] TX-100 0.2ml / L rabbit anti-human complement Clq antiserum 300ml / L

[0025] 1.3 Applicable instruments

[0026] Semi-automatic and fully automatic biochemical analyzers.

[0027] 1.4 Analytical method

[0028] Immunoturbidimetry.

[0029] 1.5 Sample Requirements

[0030] Use human serum, store at 2-8°C, test within 24 hours, if it cannot be tested within 24 hours, store at -20°C for 4 months, hemolysis and severe lipemia (such as chylus) samples cannot be used.

[0031] 1.6 Performance requirements

[0032] 1.6.1 Reagent Appearance

[0033] R1: colorless clear transparent liquid;

[0034...

Embodiment 2

[0050] Example 2 Experimental method

[0051] 2.1 Detection conditions

[0052] 2.1.1 Biochemical analyzer (hereinafter referred to as instrument)

[0053] Hitachi 7060 automatic biochemical analyzer.

[0054] 2.1.2 Working environment temperature

[0055] Room temperature 15-32°C.

[0056] Indoor humidity 45-85% RH.

[0057] 2.1.3 Main measurement parameters and operation steps

[0058] Temperature 37°C; main wavelength 340nm (secondary wavelength 700nm), sample or calibrator 4μl; 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 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.

[0061] Result calculation:

[0062]

[0063] 2.1.4 Calibrator

[0064] The calibrator used in this standard is the standard product produced by Nanfang Reagent Factory i...

Embodiment 3

[0103] The preparation of embodiment 3 reagents

[0104] 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)

[0105]

[0106] 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.

[0107] Result instrument calculation:

[0108]

[0109] Through the determination of the above samples, the figure 1 Dose-response curves of complement Clq concentration and absorbance are shown.

[0110] ( figure 1 The curve is based on the OD value with a wavelength of 340nm, and the curve with a wavelength of 700nm is for reference only. )

[0111]

[0112] pass figure 1 , measured the serum of 500 cases of normal people, the results are described ...

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Abstract

The invention belongs to the field of biological engineering, and provides a kit for detecting the concentration of a complement Clq in human serum by an immune transmission turbidimetry and an immune scattering turbidimetry. The kit solves the technical problems that an immune diffusion method and an enzyme-linked immune sorbent assay (ELISA) double antibody sandwich technology for measuring theconcentration of the complement Clq have complicated steps and are low in accuracy and repeatability in the prior art. The kit comprises two reagents, wherein a first reagent consists of disodium hydrogen phosphate, monopotassium phosphate, polyethylene glycol 6000 (PEG6000), ethylene diamine tetraacetic acid (EDTA)-NA2 and TX-100; and a second reagent consists of the disodium hydrogen phosphate,the monopotassium phosphate, the PEG 6000, the EDTA-NA2, the TX-100 and rabbit antihuman complement Clq antiserum. The invention also provides a method for detecting the concentration of the complement Clq in the human serum by using the kit. The kit and the method for measuring the concentration of the complement Clq have simple and convenient steps, are high in accuracy and repeatability and are used for automated analysis meters.

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 immunoturbidimetry and immunoscatter turbidimetry. 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 purpose of the present invention is to provide a test kit for detecting the concentration of complement Clq in human serum and a method thereof. The test kit and meth...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68G01N21/82
Inventor 刘颖成朱慧琳杨杰朱晓敏刘颖冰王泉龙刘中令
Owner 上海北加生化试剂有限公司
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