Chemiluminescent microarray chip capable of simultaneously detecting several kinds of cardiac markers and kit

Abstract

The invention discloses a chemiluminescent microarray chip capable of simultaneously detecting several kinds of cardiac markers and a kit. A matrix array group that is formed by fixing capture antibodies of detection target objects on a chip carrier is formed in a reaction region of the chemiluminescent microarray chip; and the detection target objects are several kinds of cardiac markers. Besides, the kit includes the chemiluminescent microarray chip, a second antibody solution labeled with horseradish peroxidase or alkaline phosphatase, a chemiluminescent substrate sensitive to markers, anda standard detection target object solution. With the biochip or kit detecting the several kinds of cardiac markers simultaneously, the blank of the simultaneous quantitative detection technology of the cardiac markers at the existing stage can be filled; and the requirement of detecting the cardiac markers simultaneously and rapidly in case of emergency is met. The chemiluminescent microarray chip and the kit have characteristics of multiple targets, low cost, high automation degree, high sensitivity, short detection time and great easies in operation and the like.

Description

technical field

[0001] The invention belongs to the technical field of POCT detection, relates to a biological chip, in particular to a chemiluminescent microarray chip and a kit for simultaneously detecting multiple myocardial markers. Background technique

[0002] Cardiovascular disease is a serious disease that endangers human health and life, and has become the number one killer of human health in the 21st century. Among them, acute myocardial infarction is the most common and dangerous. Changes in personal behavior and living habits have made the risk of cardiovascular disease in Chinese people surpass that of developed countries such as the United States.

[0003] Cardiovascular testing is a "bottleneck" subject in the entire cardiovascular field, because only by correctly diagnosing diseases in the shortest possible time can early detection and early treatment be possible to prevent disability and death to the greatest extent and improve patient prognosis and quality...

Examples

Embodiment 1

[0042] A biochip (or called chemiluminescent microarray chip) and kit for 5 kinds of myocardial markers, the preparation method of said biochip and kit comprises the following steps:

[0043] (1) Preparation of biochip:

[0044] The carrier of the biochip in this embodiment is a 96-well plate made of polystyrene. Each well on the carrier can independently form a reaction zone, and each reaction zone contains a set of capture antibodies immobilized on the chip carrier by detection targets (cTnI, MYO, CK-MB, D-Dimer and NT-proBNP). The above-mentioned matrix array, the number of columns of the matrix is ​​3, and each detection target antibody can be spotted with 3 repeated spots, which makes the measurement results more intuitive and accurate. Each reaction area also contains a positive control array and a negative control array, and the number of columns is 3. The specific layout is as figure 1 shown.

[0045] Before sample application, the surface of the 96-well plate is f...

Embodiment 2

[0051] The specific detection steps of the present invention for simultaneous detection of cTnI, MYO, CK-MB, D-Dimer and NT-proBNP in blood samples:

[0052] (1) The kit in Example 1 was equilibrated at room temperature for 30 minutes;

[0053] (2) On the biochip, select 6 reaction wells, add the standard solution 1-6 in Example 1 respectively in each well, and the volume added is 50 μ L; then add the blood sample after dilution in other wells ( Whole blood sample and PBS buffer of Ph7.4 were mixed at a volume ratio of 1:1), and 50 μL was added to each well; then 50 μL of HRP enzyme-labeled antibody containing a concentration of 0.3 μg / mL was added to all the above reaction wells. cTnⅠ secondary antibody, anti-MYO secondary antibody labeled with HRP enzyme at a concentration of 0.4 μg / mL, anti-CK-MB secondary antibody labeled with HRP enzyme at a concentration of 0.25 μg / mL, and anti-CK-MB secondary antibody at a concentration of 0.12 μg / mL The mixed solution of the anti-D-Di...

technical field

[0001] The invention belongs to the technical field of POCT detection, relates to a biological chip, in particular to a chemiluminescent microarray chip and a kit for simultaneously detecting multiple myocardial markers. Background technique

[0002] Cardiovascular disease is a serious disease that endangers human health and life, and has become the number one killer of human health in the 21st century. Among them, acute myocardial infarction is the most common and dangerous. Changes in personal behavior and living habits have made the risk of cardiovascular disease in Chinese people surpass that of developed countries such as the United States.

[0003] Cardiovascular testing is a "bottleneck" subject in the entire cardiovascular field, because only by correctly diagnosing diseases in the shortest possible time can early detection and early treatment be possible to prevent disability and death to the greatest extent and improve patient prognosis and quality...

Examples

Embodiment 1

[0042] A biochip (or called chemiluminescent microarray chip) and kit for 5 kinds of myocardial markers, the preparation method of said biochip and kit comprises the following steps:

[0043] (1) Preparation of biochip:

[0044] The carrier of the biochip in this embodiment is a 96-well plate made of polystyrene. Each well on the carrier can independently form a reaction zone, and each reaction zone contains a set of capture antibodies immobilized on the chip carrier by detection targets (cTnI, MYO, CK-MB, D-Dimer and NT-proBNP). The above-mentioned matrix array, the number of columns of the matrix is ​​3, and each detection target antibody can be spotted with 3 repeated spots, which makes the measurement results more intuitive and accurate. Each reaction area also contains a positive control array and a negative control array, and the number of columns is 3. The specific layout is as figure 1 shown.

[0045] Before sample application, the surface of the 96-well plate is f...

Embodiment 2

[0051] The specific detection steps of the present invention for simultaneous detection of cTnI, MYO, CK-MB, D-Dimer and NT-proBNP in blood samples:

[0052] (1) The kit in Example 1 was equilibrated at room temperature for 30 minutes;

[0053] (2) On the biochip, select 6 reaction wells, add the standard solution 1-6 in Example 1 respectively in each well, and the volume added is 50 μ L; then add the blood sample after dilution in other wells ( Whole blood sample and PBS buffer of Ph7.4 were mixed at a volume ratio of 1:1), and 50 μL was added to each well; then 50 μL of HRP enzyme-labeled antibody containing a concentration of 0.3 μg / mL was added to all the above reaction wells. cTnⅠ secondary antibody, anti-MYO secondary antibody labeled with HRP enzyme at a concentration of 0.4 μg / mL, anti-CK-MB secondary antibody labeled with HRP enzyme at a concentration of 0.25 μg / mL, and anti-CK-MB secondary antibody at a concentration of 0.12 μg / mL The mixed solution of the anti-D-Di...

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