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A kit and method for detecting the concentration of complement clq in human serum

一种检测人、浓度的技术，应用在生物工程领域，能够解决准确性不高、补体Clq浓度过程复杂等问题，达到准确性高、技术进步显著、过程简单方便的效果

Active Publication Date: 2016-05-04
SHANGHAI BEIJIA BIOCHEMICAL REAGENT CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

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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  • A kit and method for detecting the concentration of complement clq in human serum
  • A kit and method for detecting the concentration of complement clq in human serum
  • A kit and method for detecting the 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] R2: Pale yellow clear li...

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] 1 #

[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 desc...

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Abstract

The invention belongs to the field of bioengineering, and provides a kit for detecting the concentration of complement Clq in human serum by immunotransmission turbidimetry and immune scattering turbidimetry, and solves the problem of using immunodiffusion method and ELISA double-antibody sandwich technology in the prior art. Complement C1q concentration steps are complicated, the accuracy is not high, and the repeatability is poor, including two reagents, the first reagent is composed of disodium hydrogen phosphate, potassium dihydrogen phosphate, PEG6000, EDTA-NA2 and TX-100, the second The reagent consists of disodium hydrogen phosphate, potassium dihydrogen phosphate, PEG6000, EDTA-NA2, TX-100 and rabbit anti-human complement C1q antiserum. The present invention also provides a method for detecting the concentration of complement Clq in human serum by using the above-mentioned kit. The kit and method of the present invention are simple and convenient for measuring the concentration of complement Clq, with high accuracy and good repeatability, and are suitable 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 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 SHANGHAI BEIJIA BIOCHEMICAL REAGENT CO LTD
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