Whole blood C-reactive protein detection kit

A reactive protein and kit technology, applied in the field of medical immunization, can solve the problems of increasing the workload of inspection staff, increasing the pain of subjects, prolonging the detection time, etc., to achieve batch automatic analysis, enhance detection sensitivity, and broaden the linear range Effect

Inactive Publication Date: 2018-07-20
SONOSCAPE MEDICAL CORP
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For ordinary CRP, the detection range is generally 3-200mg / L, with a wide linear range but low sensitivity; while for ultra-sensitive CRP, the detection range is 0.3-100mg / L, although the sensitivity is high, high-concentration CRP cannot be measured
[0005] Although the CRP detection kits disclosed in patent CN105158476 and patent CN101769932 can guarantee high sensitivity and wide detection range at the same time, the detection samples are all serum or plasma. It is not suitable for some special patients who have difficulties in blood collection, such as children, especially infants, and people with extensive burns and scalds.
In addition, the pretreatment of the whole blood sample increases the workload of the inspection staff on the one hand, and also limits the direct automatic analysis of the sample to be tested, prolonging the detection time

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] The preparation of embodiment 1 whole blood full range CRP detection kit

[0032] Reagent R1 components: containing 0.2% SDS (sodium dodecyl sulfonate), 4% PEG6000 (polyethylene glycol 6000), 200mM sodium chloride, 2% BSA (bovine serum albumin), 1% glucose, 0.1 % Triton X-100 (Triton 100) and 0.1% sodium azide in Tris buffer. Tris buffer concentration is 100 mM, pH is 7.4.

[0033] Preparation of reagent R2: use goat anti-human CRP polyclonal antibody to sensitize small latex particles with an average particle size of 70 nm, and rabbit anti-human CRP monoclonal antibody to sensitize large latex particles with an average particle size of 250 nm. The two kinds of sensitized latex particles are ultrasonically resuspended after centrifugation and rinsing, and diluted in the second buffer solution, stabilizer, surfactant and preservative according to a certain concentration for later use. The above two sensitizing latex suspensions are mixed according to a certain ratio to...

Embodiment 2

[0037] Example 2 Determination of C-reactive protein

[0038] Detection tool: Kaili semi-automatic specific protein analyzer

[0039] Analysis method: rate nephelometry; instrument detection wavelength is 670nm.

[0040] Calibration method: Mix 10ul of calibrator with different concentrations (6 concentration gradients: 0mg / L, 5mg / L, 22.5mg / L, 44mg / L, 175mg / L, 350mg / L) with 400ul R1 and place it in the machine Add 40ul R2 to the detection channel, read the lowest scatter value A1 in the 10th second, and read the scatter value A2 after the 60th second, then calculate the scatter difference of A2 minus A1 (that is, the final astigmatism value). Subsequently, utilize spline function (Spline) to carry out the calculation and drawing of calibration curve to calibrator concentration and scattering value, standard curve is as follows figure 1 shown.

[0041] Whole blood sample test method: Take 10ul of whole blood sample, mix it with 400ul R1 and place it in the detection channel,...

Embodiment 3

[0043] Embodiment 3 kit accuracy analysis of the present invention

[0044] Take two serum calibrators traceable to international standards, including one high value and one low value. Utilize the reagent of the present invention and the specific protein instrument of our company to test relevant samples, repeat the detection 10 times for each sample, and compare with the target value of the calibrator. The results are shown in Table 1:

[0045] Table 1 Accuracy test results

[0046]

[0047]

[0048] As can be seen from the data in Table 1, the average value of C-reactive protein measured by the kit provided by the invention is very close to the target value, the accuracy of low-value sample detection is 98.36%, and the accuracy of high-value sample detection is 99.21%, indicating that this The inventive kit has high accuracy.

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Abstract

The invention relates to the field of medical immunology, and especially relates to a whole blood C-reactive protein detection kit. The kit includes a reagent R1 and a reagent R2, and the reagent R2 uses a C-reactive protein antibody to respectively sensitize latex particles having different particle sizes. Results show that the kit has the advantages of wide linear detection range, high sensitivity, good accuracy and good precision. Compared with existing human whole blood CRP turbidity kits, the kit in the invention widens the linear range, and especially enhances the detection sensitivity to low-value samples. The peripheral blood or venous blood whole blood of a patient can be used as a test sample, so the blood sampling pains of patients and especially newborns and burn patients are reduced, and true whole-blood and full-range detection of clinically hypersensitive common CRP samples is achieved.

Description

technical field [0001] The invention relates to the field of medical immunity, in particular to a whole blood C-reactive protein detection kit. Background technique [0002] C-reactive protein (CRP) is an acute phase protein synthesized by liver cells, with a molecular weight of about 115KD, and it exists in trace amounts (about 0.58mg / L) in the serum of healthy people. When the body is subjected to inflammatory stimuli such as microbial invasion or tissue damage, under the induction of cytokines such as interleukin and tumor necrosis factor, the expression of CRP will increase sharply up to more than 1000 times. And as the body recovers, its content will quickly return to normal. Therefore, the clinical detection of conventional concentration of CRP has broad significance for the identification of bacterial or viral infection, cancer and other major diseases, the progress, prognosis and evaluation of therapeutic effects. Compared with conventional CRP, the clinical signif...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/544G01N33/543
CPCG01N33/543G01N33/544G01N33/68G01N33/54313G01N2333/4737
Inventor 柴凡冯剑军刘兵曹佳强
Owner SONOSCAPE MEDICAL CORP
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