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Procalcitonin light-initiated chemiluminescence immunoassay kit and preparation method thereof

A photo-excited chemiluminescence and procalcitonin technology, applied in the field of procalcitonin photo-excited chemiluminescence immunoassay kits, can solve the problems of EIA sensitivity limitation, easy inactivation of enzyme activity, radionuclide contamination, etc. Wide detection range, high sensitivity, and easy operation

Active Publication Date: 2015-07-01
GUANGZHOU DARUI BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The most commonly used labeled immunoassay techniques are radioimmunoassay (RIA) and enzyme immunoassay (EIA), but there are some defects in practical application: RIA radionuclide pollution, short shelf life of markers; EIA enzyme activity It is easy to inactivate, resulting in low sensitivity, and the macromolecular labeling of enzymes is easy to affect the spatial structure of the labeled substance, which limits the improvement of EIA sensitivity

Method used

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  • Procalcitonin light-initiated chemiluminescence immunoassay kit and preparation method thereof
  • Procalcitonin light-initiated chemiluminescence immunoassay kit and preparation method thereof
  • Procalcitonin light-initiated chemiluminescence immunoassay kit and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1 Preparation kit

[0028] Preparation of receptor microspheres coated with anti-procalcitonin monoclonal antibody: add 0.2 mg of PCT monoclonal antibody to a centrifuge tube with a filter membrane, centrifuge at 8 000 r / min for 5-6 min, and use labeling buffer After repeated washing 6 times with the antibody solution (0.13mol / L, pH 8.0PBS), 1 mg of receptor microspheres, 10 μL of 25 mg / mL NaBH3CN (prepared with labeling buffer), 1.25 μL of 10% Tween-20 were added to the antibody solution. The volume of labeling buffer was supplemented to 200 μL, and the reaction was shaken at 37°C for 48 hours in the dark. Add 10 μL of 65 mg / mL L CMO (prepared with 0.8M NaOH) to block unbound sites, incubate at 37°C for 1 hour in the dark, and then centrifuge and wash to obtain antibody-linked receptor microspheres, which are diluted for later use.

[0029] PCT calibrator: use standard buffer (50mmol / L Tris-HCl, 1.5%BSA, 0.9%NaCl, 0.05%Proclin-300, 0.01%Tween-20, pH7.8) to pre...

Embodiment 2

[0041] Example 2 Evaluation test

[0042] Reagents: PCT calibrators, receptor microspheres coated with anti-procalcitonin monoclonal antibody, biotinylated anti-procalcitonin monoclonal antibody, streptavidin- donor microspheres.

[0043] Detection method: add calibrator, receptor microspheres coated with anti-procalcitonin monoclonal antibody, and biotinylated anti-procalcitonin monoclonal antibody into the wells of white opaque plates, incubate at 37°C with shaking 15min, then add 175μL of streptavidin-based donor microspheres, incubate at 37°C with shaking for 15min, detect the signal value on the AlphaScreen / Lisa detector, and calculate the PCT content of the tested sample from the standard curve, the unit is ng / ml, at the same time judge the negative and positive of the sample according to the S / CO value, and finally print the test report.

[0044] 1. Detection of linear range

[0045] The blood samples of 405 normal people were tested at 3 clinical trial sites, and 9...

Embodiment 3

[0067] Example 3 Clinical comparative test

[0068] Reagents: PCT calibrators, receptor microspheres coated with anti-procalcitonin monoclonal antibody, biotinylated anti-procalcitonin monoclonal antibody, streptavidin- donor microspheres.

[0069] Sample source: 126 clinical serum samples were from systemic bacterial, fungal and parasitic infections, systemic inflammatory response syndrome, sepsis, acute and chronic pneumonia, acute pancreatitis, active hepatitis, trauma, etc. 405 were healthy individuals.

[0070] Serum samples were tested by PCT-TRFIA and the chemiluminescence immunoassay kit (ILMA) of BRAHMS company abroad. The correlation analysis was carried out by SPSS17.0 software. There is a significant correlation between the detection results of , and the correlation equation is Y AlphaLisa =1.155X ILMA -0.354; Comparing the detection performance of the two methods, it can be seen that there is no significant difference between the photo-excited chemiluminescence...

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Abstract

The invention discloses a procalcitonin light-initiated chemiluminescence immunoassay kit and a preparation method thereof. The procalcitonin light-initiated chemiluminescence immunoassay kit disclosed by the invention is composed of a white opaque 96-pore plate, a procalcitonin calibrating product, a receptor microsphere coated with an anti-procalcitonin monoclonal antibody, a biotinylated anti-procalcitonin monoclonal antibody, and a streptavidin biotinylated donor microsphere. The procalcitonin light-initiated chemiluminescence immunoassay kit disclosed by the invention has the advantages of rapidness, high sensitivity, wide measuring range, simplicity in operation, and the like, and has higher sensitivity and wider detectability in comparison with an enzyme immunoassay, can be used for diagnosing and identifying individual infectious diseases, and has an application value.

Description

technical field [0001] The invention relates to the field of diagnostic testing kits, in particular to a procalcitonin photo-excited chemiluminescence immunoassay kit. Background technique [0002] Bacteria or viruses invade through the wound surface, respiratory tract, urinary tract, and digestive tract. In severe trauma, decreased systemic resistance, imbalance of normal flora in the body, bacterial translocation, etc., can cause serious infection symptoms in the body, which is a clinical Various complications, such as sepsis, septic shock, multiple organ dysfunction syndrome (MODS), and even the main cause of death. According to statistics from the United States in 2001, there were 751,000 cases of severe sepsis in that year, and the average mortality rate was as high as 28.5%, which means that the number of deaths in that year was nearly three times that of AIDS (acquired immunodeficiency syndrome, AIDS). . Early diagnosis and timely treatment of infectious diseases an...

Claims

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Application Information

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IPC IPC(8): G01N33/577
CPCG01N33/577
Inventor 吴英松徐伟文郝文波
Owner GUANGZHOU DARUI BIOTECH
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