Complement C3 detection method

A detection method and complement technology, applied in the biological field, can solve problems such as inaccurate diameter measurement, large variation, and difficult observation of results

Inactive Publication Date: 2015-08-12
王贤俊
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The one-way immunodiffusion method is time-consuming and highly variable, the results are not easy to observe, and the measurement diameter is inaccurate; the ELISA method has poor repeatability, is prone to false positives, and is most affected by temperature and time; the immunoturbidimetric method is relatively better than the previous two methods , but has the disadvantage of requiring a large amount of antiserum

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0013] The method of the present invention is formulated into a kit, and the performance is compared with that of a commercially available kit:

[0014] The concrete composition that reagent of the present invention is made into kit is as follows:

[0015]

[0016]

[0017] Use of complement C3 detection reagents:

[0018] 1) Detection instrument: a biochemical analyzer with a wavelength of 340nm and a constant temperature device at 37°C.

[0019] 2) Sample to be tested: fresh non-hemolyzed serum, stable for seven days at 2-8°C. It can be stable for one month at -20°C.

[0020] 3) Specific testing procedures:

[0021]

[0022] 4) Calculation result: complement C3 concentration (g / L) = ΔAT / ΔAS × Cs

[0023] ΔAT: Absorbance of sample tube

[0024] ΔAS: Calibration tube absorbance

[0025] Cs: calibrator concentration value

[0026] 5) Reference value range: 0.9~1.5g / L

[0027] 6) Precision: intra-assay CV≤4%; inter-assay relative range ≤6%.

[0028] 7) Accuracy...

example 1

[0041] Example 1 Reagent of the present invention compares with the performance index of commercially available reagent A:

[0042] 1) Determination of precision: 20 consecutive samples were taken for measurement, and the mean, standard deviation and coefficient of variation of the measured values ​​were calculated. CV = SD X ‾ × 100 %

[0043] Table 1 Precision test results

[0044]

[0045]

[0046] The coefficient of variation CV is usually used to measure the precision of a determination method, the smaller the CV value, the better the precision of the result of the determination method. For clinical chemistry tests, a method precision with a CV of less than 5% is generally accepted as acceptable. The CV value of the reagent of the present invention in Table 1 is less than that of commercially available reagent A, showing that the precision of t...

example 2

[0051] Example 2 Reagent of the present invention is formulated into kit related performance evaluation

[0052] 1) Determination of precision: 20 consecutive samples were taken for measurement, and the mean, standard deviation and coefficient of variation of the measured values ​​were calculated. CV = SD X ‾ × 100 %

[0053] Table 3 precision test results

[0054]

[0055]

[0056] The coefficient of variation CV is usually used to measure the precision of a determination method, the smaller the CV value, the better the precision of the result of the determination method. For clinical chemistry tests, a method precision with a CV of less than 5% is generally accepted as acceptable. The CV value in Table 3 is less than 3%, indicating that the method of the present invention has excellent precision.

[0057] 2) Measurement of accuracy: use the same ...

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Abstract

The invention provides a complement C3 detection method, which is based on the latex-enhanced immune turbidimetry. In the detection method, two liquid reagents (reagent R1 and reagent R2) are used, the reagent R1 comprises a phosphate buffer solution (pH=6), tween 20, disodium ethylene diamine tetraacetic acid (EDTA-2Na), sodium chloride, polyethylene glycol (PEG6000) and sodium azide; and the reagent R2 is a solution containing complement C3 latex particles. The detection method is characterized in that chemical crosslinking is used, and through the water-soluble carbodiimide (EDC) and N-hydroxy succinimide (NHS), the goat anti-human complement C3 antiserum can carry out covalent crosslinking with carboxylated polystyrene latex to form a complement C3 latex reagent. The complement C3 latex reagent has the advantages of high sensitivity, strong specificity, and simple preparation, and thus is worthy to promote and use.

Description

Technical field: [0001] The invention belongs to the field of biotechnology, and provides a complement C3 detection method, which has the characteristics of high sensitivity, strong specificity and simple reagent preparation, and is worthy of further popularization and use. Background technique: [0002] C3 is the complement component with the highest content in serum. It is a kind of β globulin synthesized by macrophages, monocytes, lymphoid tissue, bone marrow, peritoneum and liver. Under the action of C3 convertase, it is cracked into C3a and C3b. It plays an important role in both the classical and alternative pathways of complement activation. [0003] C3 is the one with the highest content among the components of complement, and is the most important link in the complement activation pathway, so the determination of its content is very important. The increase and decrease of C3 are basically consistent with the total complement activity, but more sensitive. About 70%...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N21/33
CPCG01N33/6854G01N21/33
Inventor 王贤俊郑蓓蕾郭二豪江新涛
Owner 王贤俊
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