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

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...

AI Technical Summary

Problems solved by technology

[0010] Using other existing technologies or (1) the structure of the chip is too complicated to precisely regulate the reaction, and there is a large pollution in the cleaning steps and liquid conduction; or (2) multiple liquid chambers and solid phase carriers are used (magnetic beads and fluorescent microspheres are used at the same time), the overall design of the reagent is too complicated; or (3) the immunoreaction adopts a heterogeneous reaction, which means that the solid phase carrier reacts with the surface of the solid phase carrier, the reaction is insufficient, and the sensitivity and Repeatability is limited

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