MULTIASSAY IMMUNOCHROMATOGRAPHIC CHIP

• US 20130157380A1
• Filed: 08/18/2011
• Published: 06/20/2013
• Est. Priority Date: 08/19/2010
• Status: Abandoned Application

• Current Assignee: Institute Of Microbiology And Epidemiology Academy Of Military Medical Sciences
• Original Assignee: Institute Of Microbiology And Epidemiology Academy Of Military Medical Sciences
• Inventors: Zhou, Lei, Yang, Ruifu, Guo, Zhaobiao
• Application Number:
  US13/635,627
• Publication Number:
  US 20130157380A1
• Time in Patent Office: Days
• US Class Current: 436/501
• CPC Class Codes: G01N 33/5302 Apparatus specially adapted for immunological test procedures
• CPC Class Codes: G01N 33/54388 based on lateral flow
• CPC Class Codes: G01N 33/558 using diffusion or migration of antigen or antibody immunochromatographic test strips G01N33/54387
• CPC Class Codes: Y10T 156/1052 with cutting, punching, tearing or severing

Abstract

A multiassay immunochromatographic chip comprises: a viscous bottom lining (1), a sample pad (2), a bonding pad (3), an analysis membrane (4) and a water absorption pad (5). The bonding pad (3) fixedly has a plurality of assay binding substances (6) and a control binding substance (7), the assay binding substance (6) is formed by joining a tracer (8) and a liquid-phase detection probe (9), the control binding substance (7) is formed by joining the tracer (8) and a liquid-phase control probe (11); the analysis membrane (4) is provided with a detection matrix unit (12), each detection matrix unit comprises a detection zone (13) and a control zone (14), wherein the detection zone (13) consists of a plurality of solid phase detection probes (15) and the control zone (14) consists of a solid phase control probe (16). Organic integration of immunochromatographic reaction modes and chip assay matrix settings enables high throughput assay of multiple target substances with one sample load.

Claims

1. A multiassay immunochromatographic chip, comprising:

a laminating card [1], a sample pad [2], a conjugate pad [3], an analysis membrane [4], and an water absorption pad [5],

wherein, several kinds of test conjugates [6] and a control conjugate [7] are fixed to the conjugate pad [3];

each kind of the test conjugates [6] is formed by conjugating a marker [8] with a corresponding kind of liquid-phase test probe [9], and is in specific one-to-one correspondence to each kind of detected targets [10];

the analysis membrane [4] is provided with a test array unit [12];

the test array unit [12] comprises a test zone [13] and a control spot [14];

the test zone [13] comprises various kinds of solid-phase test probes [15], and each kind of the solid-phase test probes [15] on the test zone [13] is positioned fixedly at a well-defined position and corresponds to the specific detection of a certain kind of detected target [10].

2. The multiassay immunochromatographic chip according to claim 1, wherein, the laminating card [1] is an object made of a rigid material and coated with a pressure sensitive adhesive on one surface, and the object is PVC plate.

3. The multiassay immunochromatographic chip according to claim 1, wherein, the sample pad [2] is an object that has a large bed volume and a uniform microstructure, and the object is selected from water absorption paper, cellulose membrane, fiber glass, non-woven fabric, or blood filtering membrane.

4. The multiassay immunochromatographic chip according to claim 1, wherein, the conjugate pad [3] is an object that has a large bed volume and a uniform microstructure, and the object is selected from glass fiber, polyester membrane, or non-woven fabric;

the control conjugate [7] is formed by conjugating the marker [8] with a liquid-phase control probe [11], and can control whether the immunochromatographic analysis is in a normal state.

5. The multiassay immunochromatographic chip according to claim 1, wherein, the analysis membrane [4] is an object that has a uniform microstructure, and the object is selected from nitrocellulose membrane or nylon membrane;

the control spot [14] comprises a solid-phase control probe [16], and the solid-phase control probe [16] on the control spot [14] is positioned fixedly at a well-defined position to control whether the entire immunochromatographic analysis is in a normal state.

6. The multiassay immunochromatographic chip according to claim 1, wherein, the water absorption pad [5] is an object that has a large bed volume, and the object is selected from water absorption paper or cellulose membrane.

7. The multiassay immunochromatographic chip according to claim 1, wherein, each detected target [10] corresponds to two test probes, wherein, one of the test probes is fixed as the solid-phase test probe [15] on the analysis membrane [4], and the other test probe as the liquid-phase test probe [9] is conjugated with a marker [8] to form the test conjugate [6] fixed on the conjugate pad [3].

8. The multiassay immunochromatographic chip according to claim 1, wherein, each kind of the solid-phase test probes [15] is positioned fixedly at well-defined position on the test zone [13] of the analysis membrane [4], and corresponds to specific detection of a certain kind of detected target [10] individually;

each kind of the solid-phase test probes [15] is specifically immunologically reacted with a certain kind of detected target [10] and the corresponding kind of liquid-phase test probe [9] of the test conjugate [6], so that the amount of the marker [8] bonded at the well-defined position where this kind of solid-phase test probe [15] is located is changed by the immunologic reaction, and thereby the existence and concentration of this kind of detected target [10] is revealed by the amount of the marker [8].

9. The multiassay immunochromatographic chip according to claim 5, wherein, the solid-phase control probe [6] bonds directly with the liquid-phase control probe [11] of the control conjugate [7] to control whether the immunochromatographic analysis is in a normal state.

10. The multiassay immunochromatographic chip according to claim 1, comprising a sandwich immunochromatographic chip, an indirect immunochromatographic chip, and a competitive immunochromatographic chip;

wherein, the sandwich immunochromatographic chip comprises a double-antibody sandwich immunochromatographic chip for antigen detection and a double-antigen sandwich immunochromatographic chip for antibody detection;

the indirect immunochromatographic chip is used for detection of specific antibodies in blood serum sample, with the test conjugate formed by conjugating a marker with a secondary antibody of detected antibodies;

the competitive immunochromatographic chip is used for the detection of small-molecule substances, such as a hapten that has only one antigenic determinant.

11. A method for preparation of the multiassay immunochromatographic chip according to claim 1, comprising the following steps:

A. Preparation of a conjugate pad [3];

mixing a control conjugate [7] with various kinds of test conjugates [6] to obtain a mixture of conjugates, applying the mixture on a piece of fiber glass, polyester membrane, or non-woven fabric that is used as the conjugate pad [3], and drying the resultant for future use;

B. Preparation of an analysis membrane [4];

adding respectively various kinds of antigens that serve as the solid-phase test probes [15] and an antibody that serves as the solid-phase control probe [16] in the form of round spots on a nitrocellulose membrane or a nylon membrane at well-defined, fixed and addressable positions, to form a test zone [13] and a control spot [14] respectively, and thereby form a test array unit [12]; and

drying it for future use, wherein, in the test array unit [12], each kind of the solid-phase test probes [15] in one-to-one correspondence to the detected targets [10] is fixed at a well-defined position, and only one solid-phase control probe [16] is required, which also has a well-defined fixed position.

C. Laminating and cutting;

laminating the sample pad [2], conjugate pad [3], analysis membrane [4], and water absorption pad [5] in sequence to a PVC plate serving as the laminating card [1], while ensuring overlapping relation between each parts;

the finished immunochromatographic chip can be used directly or loaded into a plastic cartridge for use, so as to obtain the immunochromatographic chip in the invention.

12. A method for preparation according to claim 11, further comprising:

preparing a plurality of the test array units [12] on the analysis membrane [4] in the step B, and further cutting the PVC plate into separate and usable finished products at the break points [17] between the test array units [12] followed in the step C.

13. A method for biological target detection with the multiassay immunochromatographic chip according to claim 1, comprising:

A. Adding sample;

adding a liquid sample or pretreated liquid sample in droplets onto the sample pad [2] of the immunochromatographic chip according to any one of claims 1-10;

B. Immunochromatographic reaction;

standing for several minutes, till the immunochromatographic reaction is completed, wherein in the immunochromatographic reaction process, specific immunologic reactions occur among the test conjugates [6], the detected targets [10], and the solid-phase test probes [15], and the bonded amount of the marker [8] is changed at the well-defined fixed positions on the analysis membrane [4], and the bonded amount of the marker [8] at a certain position directly reflects the existence or concentration of a certain kind of detected target [10];

C. Result judgment;

judging the result directly by visual examination for a color marker [8], or judging the result with instruments for a marker [8] that generates an optical, electrical, or magnetic signal, thereby obtaining a qualitative and quantitative detecting result for a certain kind of detected target [10] corresponding to the well-defined fixed position.