Kit and method for detecting procalcitonin

A procalcitonin and kit technology, applied in the field of immunoassays, can solve problems such as unstable markers, difficulty in ensuring repeatability of results, and limited internal surface area, so as to improve detection sensitivity and accuracy and avoid reduction in luminous efficiency , the effect of large antibody coating surface area

Active Publication Date: 2012-02-22
SHENZHEN GOLDSITE DIAGNOSTICS
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  • Abstract
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Problems solved by technology

[0010] The above methods for detecting PCT have the following disadvantages: 1) The luminescence efficiency of direct labeling antibodies (or antigens) with acridinium ester or isoluminol is low, and the markers are unstable. The luminescence process is completed in less than 1s), it is difficult to guarantee the repeatability of the results, and a complex and accurate in-situ detection mechanism is required, which increases the difficulty of the design of the detection instrument; 2) The reaction tube or the well of the luminescent plate is directly coated as a fixed phase, which has a limited internal surface area and cannot achieve a higher

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  • Kit and method for detecting procalcitonin
  • Kit and method for detecting procalcitonin
  • Kit and method for detecting procalcitonin

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Experimental program
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Effect test

preparation example Construction

[0036] 1. Preparation of magnetic particle-conjugated antibodies

[0037] The magnetic particle-coupled antibody in the kit is used as a stationary separation phase. The antibody in the magnetic particle-coupled antibody is procalcitonin monoclonal antibody. The concentration of the magnetic particle used for coupling is 1-10 mg / mL. The concentration of the linked antibody is 1-5mg / mL, the size of the magnetic particle is 200nm-5μm, and the active group on the surface of the magnetic particle is -COOH or -NH 2 , the magnetic particles are activated by chemical cross-linking reagents, so that they can be separated from the -NH of procalcitonin monoclonal antibody 2 Or the -COOH group is covalently bonded to form a stable magnetic particle conjugate. In this embodiment, the magnetic particle whose active group is -COOH is the first choice. Under the action of procalcitonin monoclonal antibody -NH 2 The group forms an amide bond and is covalently bonded. The magnetic particle-c...

Embodiment 2

[0059]The detection method of procalcitonin, the detection process can adopt a one-step method or a two-step method reaction mode: in the one-step method, the working solution, the serum to be tested and the magnetic particle-coupled antibody solution are added to the reaction tube at the same time, and after a certain period of reaction , magnetic separation and washing, adding a luminescent substrate, luminol or isoluminol emits photons under the catalysis of HRP, and the luminescent signal is detected by the detector. In the two-step method, the serum to be tested and the working solution are first added to the reaction tube to react for a certain period of time, then the magnetic particle-coupled antibody solution is added, after a certain period of time, the magnetic separation and washing are added, and the luminescent substrate, luminol or isoglucose, is added. Under the catalysis of Minol HRP, photons are emitted, and the luminescent signal is detected by the detector. ...

Embodiment 3

[0067] Embodiment three: the methodological evaluation of kit of the present invention

[0068] 1. Accuracy

[0069] Accuracy measurement refers to adding a known amount of PCT standard substance to normal human serum samples, and comparing the measured concentration value with the added theoretical value to calculate the recovery rate of PCT. The test results are as follows:

[0070]

[0071] 2. Precision

[0072] Three specimens with different concentrations were selected, and the measurements were repeated 20 times according to the method of the present invention. According to the measurement results of 20 times, calculate the average deviation C.V.% value.

[0073]

[0074] 3. Analytical sensitivity

[0075] Analytical sensitivity is defined as: Refers to the quantity that can be distinguished from zero dose in a statistical sense. Repeat 20 times to measure the zero dose point, calculate the mean (X) and standard deviation (SD), and the calculated concentration...

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Abstract

The invention discloses a kit for detecting procalcitonin. The kit comprises a luminous substrate, a magnetic particle coupling antibody, a biotinylation antibody and an enzyme marker, wherein the enzyme marker is horseradish peroxidase labeled streptavidin; and the antibody is a procalcitonin monoclonal antibody. In the kit, the magnetic particle coupling antibody is used as a fixed separation phase, a biotin-streptavidin multistage amplification system is adopted, the luminous substrate is catalyzed by using horseradish peroxidase serving as catalytic enzyme to emit light, and the detectionsensitivity and accuracy of reagents are improved.

Description

technical field [0001] The invention relates to the field of immune analysis, in particular to a kit for detecting procalcitonin and a detection method for procalcitonin. Background technique [0002] In recent years, the pathological and physiological processes of systemic infection have been better understood, and intensive supportive treatment has also been significantly improved. However, systemic infection and its complications are still the leading cause of death in non-cardiac intensive care units, the United States Approximately 500,000 patients with systemic infections are diagnosed each year, and approximately 40% eventually die. Therefore, there is a need for a marker that can more effectively reflect the degree of inflammatory damage caused by infection. Procalcitonin (abbreviated as PCT) is a new indicator with high sensitivity and specificity. It is significantly elevated when bacterial or fungal infection is combined with severe systemic reactions or organ hy...

Claims

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Application Information

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IPC IPC(8): G01N21/76G01N33/577G01N33/535
Inventor 陆春郑筱雯陈明峰
Owner SHENZHEN GOLDSITE DIAGNOSTICS
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