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Method for quantitatively detecting SAA/PCT/CRP (Serum Amyloid Protein A/Procalcitonin/C-Reactive Protein) simultaneously

A technology for quantitative detection and detection of test strips, applied in the field of medical detection, can solve problems such as the increase of CRP content, and achieve the effects of low cost, high detection sensitivity, simple and stable detection method

Inactive Publication Date: 2016-05-04
天津中新科炬生物制药股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, during inflammatory disease activity in autoimmune diseases, PCT levels are not elevated, but CRP levels are elevated

Method used

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  • Method for quantitatively detecting SAA/PCT/CRP (Serum Amyloid Protein A/Procalcitonin/C-Reactive Protein) simultaneously
  • Method for quantitatively detecting SAA/PCT/CRP (Serum Amyloid Protein A/Procalcitonin/C-Reactive Protein) simultaneously

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] 1 main material

[0049] 1.1 Biological material SAA, PCT and CRP paired antibodies were purchased from Finland HYTEST company; goat anti-mouse IgG: self-made; chloroauric acid: product of Sigma company; NC membrane: product of Sartorius company; bovine serum albumin (BSA), polyethylene glycol Alcohol PEG20000: product of Sigma. Other commonly used reagents are analytical reagents.

[0050] 1.2 The clinical samples were obtained by the company in relevant hospitals, a total of 120 samples, including 60 serum samples from patients diagnosed with inflammatory infection type, the detection value of SAA was 5-200mg / L, the detection value of PCT was 0.5-20mg / L and 1-200mg / L L. 60 normal human serum samples, SAA, PCT and CRP values ​​were not detected.

[0051] 1.3 Triple test card: designed by our company, and produced and provided by related companies according to requirements.

[0052] 1.4 Immunochromatography result interpretation recorder: model: NS3001, product of T...

Embodiment 2

[0067] A detection test strip for simultaneous and rapid quantitative detection of SAA / PCT / CRP is prepared by the following method, and the specific steps are:

[0068] (1) Colloidal gold labeling of SAA, PCT and CRP antibodies The colloidal gold solution with a diameter of 40±5nm was prepared by the chloroauric acid-trisodium citrate method, and three colloidal gold solutions were taken, and the solutions were adjusted to SAA with 0.2MK2CO3 Adjust the pH of PCT to 7.5, the pH of PCT to 6.5 and the pH of CRP to 8.0, then slowly stir the colloidal gold, SAA: add 10ug of SAA-labeled antibody to the solution per ml of solution; PCT: per ml of solution Add 15ug to add the PCT-labeled antibody to the solution; CRP: add 5ug per ml solution to add the CRP-labeled antibody to the solution, continue to stir for 30 minutes, and then add 0.5% PEG2000 and 0.5% bovine serum albumin to the final concentration (BSA) for blocking, centrifuged at 13000r / min after labeling, discarded the supern...

Embodiment 3

[0072] (1) Colloidal gold labeling of SAA, PCT and CRP antibodies The colloidal gold solution with a diameter of 40±5nm was prepared by the chloroauric acid-trisodium citrate method, and three colloidal gold solutions were taken, and the solutions were adjusted to SAA with 0.2MK2CO3 The pH of PCT was adjusted to 7.0, the pH of PCT was adjusted to 7.0 and the pH of CRP was adjusted to 7.0, then the colloidal gold was stirred slowly, SAA: add 8ug of SAA-labeled antibody to the solution per ml of solution; PCT: per ml of solution Add 18ug to add the PCT-labeled antibody to the solution; CRP: add 8ug per ml solution to add the CRP-labeled antibody to the solution, continue to stir for 30 minutes, and then add 0.5% PEG2000 and 0.5% bovine serum albumin to the final concentration (BSA) for blocking, centrifuged at 13000r / min after labeling, discarded the supernatant, and redissolved the precipitate in colloidal gold working solutions of different proportions (0.1M Tris.HCl buffer, pH...

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Abstract

The invention provides a method for quantitatively detecting SAA / PCT / CRP (Serum Amyloid Protein A / Procalcitonin / C-Reactive Protein) simultaneously. Test paper of human SAA, test paper of PCT and test paper of CRP, which are mutually independent, a triple card shell and an immunochromatographic interpretation result recorder with a corresponding judgment method and a standard curve are used for quantitatively detecting SAA / PCT / CRP simultaneously and rapidly. The detection method is simple and stable and does not require a large number of samples, thereby reducing the medical cost; and the method has high sensitivity and can rapidly and quantitatively detect SAA / PCT / CRP simultaneously.

Description

technical field [0001] The invention relates to the field of medical detection, in particular to a method for simultaneous quantitative detection of SAA / PCT / CRP. Background technique [0002] Serum amyloid A (SAA) is a highly heterogeneous acute phase reaction protein, mainly produced by liver cells, with a relative molecular weight of about 12,000. The concentration of SAA, an acute reactant, in the inflammatory response will be 1000 times higher than that in normal conditions, and the increase is higher than that of CRP. At this time, it can bind to high-density lipoprotein (HDL) through its N-terminal to replace the HDL-bound protein. Apolipoprotein A-I (ApoA-I) thus forms the SAA / HDL complex in the acute phase, and its half-life is only about 50 minutes. Studies have shown that in the case of severe inflammation, the level of SAA expression is closely related to the number of necrotic cells and tissues. The expression of SAA peaked on the 3rd day of inflammation and re...

Claims

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Application Information

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IPC IPC(8): G01N33/74G01N33/68G01N33/558
CPCG01N33/74G01N33/558G01N33/6893G01N2333/47G01N2333/4737G01N2333/585G01N2800/26
Inventor 李洲周洪锐陈逸桐张梓琪李昀地杨延瑞
Owner 天津中新科炬生物制药股份有限公司
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