Procalcitonin detection kit and preparation and detection method based on microfluidic chip

A microfluidic chip and procalcitonin technology, applied in the field of immunodetection, can solve the problems of limited sensitivity and repeatability, insufficient response, complex chip structure, etc., to reduce pollution, reduce system, and avoid processing steps. Effect

A microfluidic chip and procalcitonin technology, applied in the field of immunodetection, can solve the problems of limited sensitivity and repeatability, insufficient response, complex chip structure, etc., to reduce pollution, reduce system, and avoid processing steps. Effect

CN108387564BActive Publication Date: 2020-08-21NANJING LANSION BIOTECH CO LTD
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  • Procalcitonin detection kit and preparation and detection method based on microfluidic chip
  • Procalcitonin detection kit and preparation and detection method based on microfluidic chip
  • Procalcitonin detection kit and preparation and detection method based on microfluidic chip

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Embodiment Construction

[0040] Below in conjunction with the embodiment of the present invention further elaborates:

[0041] The procalcitonin PCT kit based on the microfluidic chip of the present invention includes a microfluidic chip and a calibrator, and the calibrator is composed of A component and B component, wherein the calibration The product diluent is the calibrator A component, the concentration is 0.5ng / mL; the calibrator B component is the PCT antigen mother solution diluted to 2 ng / mL with the calibrator diluent; The reagents are stored dry in the microfluidic chip; another PCT antibody is coated in the reaction chamber of the microfluidic chip.

[0042] In this embodiment, the preparation method of the procalcitonin PCT kit based on the microfluidic chip comprises the following steps:

[0043] (1) Microfluidic chip coating: Take PCT antibody M7402 (Beijing Zhongke Jingda Biotechnology Co., Ltd.), add it to CB buffer solution with pH 9.6, and add 25 μL to the reaction chamber of the m...

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Abstract

The invention discloses a micro-fluidic chip-based procalcitonin PCT kit, and a preparation method and a detection method thereof. The preparation method of the kit comprises the following steps: (1)micro-fluidic chip coating; (2) fluorescence microsphere labeling; (3) micro-fluidic chip assembling; (4) preparation of a calibration substance; and (5) obtaining of the micro-fluidic chip-based procalcitonin PCT kit. The prepared PCT kit adopts a double-antibody sandwich technology to achieve detection, a highly sensitive time-resolved fluorescent substance is selected as a marker, the fluorescent microsphere is labeled with an antibody, the immunoreaction of an antibody pair is used to achieve analysis and detection, and the performances of a reagent prepared in the invention reach the level of a same chemiluminescent reagent. Superfluous components are completely removed by using a cleaning solution after an immune reaction, and fluorescent microspheres combined in the immune reactionare detected and read, so the problem of the insufficient reaction of a color developing solution in the chip or an untimely reading after color development is avoided.

Description

technical field [0001] The invention belongs to the field of immune detection, and in particular relates to a procalcitonin detection kit based on a microfluidic chip and a preparation and detection method thereof. Background technique [0002] Procalcitonin (PCT), the precursor protein of calcitonin, is a prohormone composed of 116 amino acids with a molecular weight of about 13 kilodaltons. PCT is expressed in C cells of thyroid, lung and pancreas tissues, and after enzymatic decomposition, it will be decomposed into calcitonin (32 amino acids), anticalcitonin (21 amino acids) and amino terminal peptide (57 amino acids). There is only a very small amount of PCT in the blood of healthy people. After the human body is infected with bacteria, the level of PCT will increase significantly, and various tissues in the body will secrete and express PCT. The concentration of PCT can rise 2-3 hours after infection, reach the peak at 6-12 hours, maintain a high level within 48 hour...

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Application Information

Patent Timeline
21 Aug 2020
Publication
CN108387564B
IPC
G01N21/64
CPC
G01N21/6486
Inventors
许行尚; 杰弗瑞·陈