CN-121978188-A - Disposable electrochemical organism sensor detection kit

CN121978188ACN 121978188 ACN121978188 ACN 121978188ACN-121978188-A

Abstract

The invention discloses a disposable electrochemical biosensor detection kit, and belongs to the technical field of in-vitro diagnosis. The disposable in-vitro diagnosis product of the kit is divided into two large product systems of molecular detection and immunodetection, wherein the core components of the molecular detection kit are an integrated disposable H-type electrolytic tank, constant-temperature amplification buffer solution, positive control, negative control and blank control solution, and the core components of the immunodetection kit are an integrated disposable H-type electrolytic tank, detection reagent, substrate reagent, calibrator I, calibrator II, quality control product I and quality control product II. The kit takes a functionalized nuclear pore membrane as a core, the nuclear pore membrane is modified in advance to correspond to a target recognition probe according to different detection targets, is pre-sealed and fixed in an H-shaped electrolytic tank and forms an integral detection unit which is not detachable with the electrolytic tank, the sample transmembrane transmission is realized through the pressurization of a single inflation inlet, and the high-precision detection can be realized by combining an isothermal amplification or double-antibody sandwich technology and an electrochemical sensing technology.

Inventors

YAN JIZHI
ZHENG NA
ZHANG JUNTAO
ZHANG BO
LIU TIEMEI

Assignees

协和生物制药（天津）有限公司

Dates

Publication Date: 20260505
Application Date: 20260409

Claims (3)

1. A disposable electrochemical biosensor detection kit is characterized in that the whole kit is a disposable in-vitro diagnosis product and is divided into two large product systems of molecular detection and immunodetection, the molecular detection kit comprises a disposable H-shaped electrolytic tank, constant-temperature amplification buffer solution, positive control, negative control and blank control solution, the immunodetection kit comprises a disposable H-shaped electrolytic tank, detection reagents, substrate reagents, a first calibrator, a second calibrator, a first electrolyte control and a second electrolyte control, the disposable H-shaped electrolytic tank is of an integrated non-reusable H-shaped electrolytic tank structure, a functional nuclear pore membrane is pre-sealed and fixed between two mutually independent chambers of the H-shaped electrolytic tank, a non-detachable disposable detection unit is formed by the two chambers, the two chambers are wholly discarded after being used, the functional nuclear pore membrane is pre-marked with target recognition probes corresponding to detection targets, the inner walls of pore channels of the functional nuclear pore membrane are only required to be replaced in a pre-preparation stage for different detection, the recognition probes pre-marked on the functional nuclear pore membrane are required to be replaced in a pre-preparation stage, the functional nuclear pore membrane can be used for the target detection targets, the two chambers are required to be provided with a pressure-adjustable electrolyte filling system 1, and an inflation port 3 is required to be used for the whole electrolyte filling system, and an inflation port 1 is required to be provided, and an inflation port 1 is required to be used for the electrolyte.
2. The disposable electrochemical biosensor detection kit according to claim 1, wherein the functionalized nuclear pore membrane is prepared by taking a polymer membrane bombarded by heavy ions as a base material, then performing chemical etching on the base material to prepare a film containing a plurality of nanometer pore channels, performing activation treatment, pre-fixing target recognition probes corresponding to detection targets, and finally closing active sites, wherein the prepared functionalized nuclear pore membrane is pre-sealed and fixed between two chambers of a disposable H-shaped electrolytic tank, is of an integrated disposable structure with the electrolytic tank, and is disposable along with the whole electrolytic tank.
3. A disposable electrochemical biosensor detection kit according to claim 1 is characterized in that the conventional composition of each composition of the kit comprises (1) a isothermal amplification buffer solution containing Bst DNA polymerase, isothermal amplification specific primers, thermostable uracil DNA glycosidase, tris-HCl buffer solution, mgCl 2 , dNTPs and amplification accelerator, (2) a detection reagent containing a buffer system of a target specific labeled antibody and containing protective proteins and preservatives, (3) a substrate reagent containing a chromogenic substrate component and a buffer solution system, (4) a positive control containing a buffer system of a target object with a known concentration and containing protective proteins and preservatives, (5) a negative control containing a blank matrix buffer system of a target object without containing protective proteins and preservatives, (6) a blank buffer system of a target object without containing preservatives, (7) a calibrator I and a calibrator II respectively containing recombinant antigens with different concentration gradients, and (8) a substrate control containing protective proteins and preservatives I and a calibrator II respectively containing recombinant antigens in a target object concentration range.

Description

Disposable electrochemical organism sensor detection kit Technical Field The invention belongs to the technical field of in-vitro diagnosis, and particularly relates to a full-component disposable detection kit based on a functionalized nuclear pore membrane and an electrochemical sensing technology. Background The in vitro diagnosis is a core support means for early screening, accurate diagnosis, dynamic monitoring of the course of disease and personalized medication guidance of clinical diseases, and occupies more than 70% of information sources of clinical diagnosis and treatment decisions. The current clinical mainstream in-vitro diagnosis technology still has a plurality of industrial pain points which are difficult to break through, namely 1, in the field of nucleic acid molecule detection, fluorescent quantitative PCR is used as clinical gold standard, highly depends on a high-precision variable-temperature amplification instrument, has high equipment cost and poor portability, and is difficult to popularize in basic medical institutions and bedside detection scenes; the technology has the advantages that the whole flow detection period is usually more than 2 hours, the requirements of emergency treatment and on-site rapid screening cannot be met, the problems that amplified products are easy to pollute and have high false positive rate exist, 2, in the field of protein immunodetection, a chemiluminescent and enzyme-linked immunosorbent assay is the current mainstream technology, the problems that the sample consumption is large, the reaction incubation time is long, the operation steps are multiple, the anti-interference capability is weak are generally existed, the detection result is easily influenced by environment and operation factors, the stability among batches is poor, and the rapid quantitative detection beside a bed cannot be realized due to the dependence on large detection equipment, 3, an electrochemical biosensor is used as the front edge technology of fusion of biological identification and electrochemical signal conversion, the technology has the core advantages of rapid response, simple and convenient operation, trace sample detection and real-time output result, and is the key development direction in the field of in vitro diagnosis, but the existing electrochemical biosensor technology still has obvious short plates, firstly, the traditional glassy carbon electrodes and silk-screen printing electrodes are mostly adopted by the sensing carrier, the biocompatibility is poor, the specific surface area is limited, the biological identification element fixing efficiency is low, the bonding stability is poor, the detection signal to be low, the noise ratio is low, the repeatability is poor, the nuclear is and the density is Kong Moji, the electrochemical sensor is a uniform, the density is the existing electrochemical sensor device is a high, and the density is a porous channel is not realized, the detection sensitivity and the detection limit are difficult to meet the detection requirement of a clinical micro target, the prior art can only realize a single molecule or immune detection system, cannot be compatible with two large core detection scenes, has poor universality and limited clinical application scenes, and the core detection unit is not capable of realizing full-closed disposable due to the lack of a complete matched disposable kit system, has the risk of cross contamination, does not have matched reference substances, calibration materials and quality control substances, cannot meet the quality control requirement of the whole process of clinical detection, and is difficult to realize industrialization landing. Based on the detection kit, a set of electrochemical biosensor detection kit which has high detection performance, is compatible with a double system, is complete in matching, is disposable in all components and can be produced in a standardized manner is developed, and the detection kit becomes a core problem to be solved in the industry. Disclosure of Invention The invention aims to overcome the defects of the prior art, solve the core problems of high equipment dependence, long detection period, complicated operation, poor universality, repeated use cross contamination risk and insufficient detection performance of the conventional in-vitro diagnosis technology, provide a disposable electrochemical biosensor detection kit, be compatible with two systems of molecular detection and immunodetection, and realize rapid, accurate, standardized and pollution-free detection of multiple clinical targets and multiple scenes by taking a universal integrated disposable H-shaped electrolytic cell as a core detection unit. The invention aims at realizing the aim, and adopts the following technical scheme that the disposable electrochemical biosensor detection kit is a disposable in-vitro diagnosis product, is divided into two large product systems of molecular detection and immu