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Immunochromatographic detection test strip for joint detection of NSE and CEA, and preparation method and application method thereof

A technology of immunochromatography detection and combined detection, which is applied in the direction of biological testing, measuring devices, analysis materials, etc., can solve the needs of simultaneous detection of multiple markers, the sensitivity of immunochromatography detection needs to be improved, and the unfavorable accuracy of quantitative detection Sex and other issues, to achieve the effect of improving screening and diagnosis, improving sensitivity and accuracy, and simple structure

Inactive Publication Date: 2016-08-17
SHANGHAI JIAO TONG UNIV
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AI Technical Summary

Problems solved by technology

[0009] 1. Traditional enzyme-linked immunosorbent assay and mature immunochromatographic detection can only detect one marker at a time, which cannot meet the needs of simultaneous detection of multiple markers
[0010] 2. The existing detection technology, because the detection results can only be judged by visual inspection, is very subjective; therefore, only qualitative detection, semi-quantitative or inaccurate quantitative detection can be realized; the detection sensitivity of immunochromatography also needs to be improved
[0011] 3. Quantitative immunoassay technology based on photoelectric effect can only detect the light of immune particles on the surface of nitrocellulose membrane, so it is difficult to truly reflect the total amount of immune particles on the detection line, which is not conducive to the accuracy of quantitative detection

Method used

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  • Immunochromatographic detection test strip for joint detection of NSE and CEA, and preparation method and application method thereof
  • Immunochromatographic detection test strip for joint detection of NSE and CEA, and preparation method and application method thereof
  • Immunochromatographic detection test strip for joint detection of NSE and CEA, and preparation method and application method thereof

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Embodiment 1

[0042] Example 1: Preparation of a combined detection reagent strip for lung cancer markers NSE and CEA based on magnetic nanoparticles

[0043] Step 1: Preparation of the first nano-magnetic bead immunoprobe and the second nano-magnetic bead immunoprobe

[0044] Take 2mg of carboxyl magnetic beads with a particle size of 80nm and place them in 500ul activation buffer (MES, pH=5.5, 0.01M), add 5mg each of EDC and NHS successively, and activate for 30 minutes; use a magnetic separation rack to separate the magnetic beads, After the supernatant was discarded, the unreacted activator was washed with coupling buffer (BS, 0.005M, pH=9.0). Afterwards, appropriate amount of labeled antibodies of NSE and CEA were added, reacted in a shaker at 36°C for 3 hours, respectively, and blocked with blocking solution (BS buffer solution containing 8% BSA) for 1 hour, and separated magnetic beads with magnetic separation racks, respectively. After discarding the supernatant, wash with BST for ...

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Abstract

The invention discloses an immunochromatographic detection test strip for joint detection of NSE (neuron-specific enolase) and CEA (carcino-embryonic antigen). The test strip adopts a double antibody sandwich technology, and includes a backer board, and a sample pad, a binding pad, a chromatographic pad and an absorbent pad which are sequentially laid on the backer board in an overlapped manner; the binding pad is provided with a first nano-magnetic bead immune probe and a second nano-magnetic bead immune probe; the first nano-magnetic bead immune probe includes a carboxyl nano-magnetic bead and an anti-NSE monoclonal antibody, and the second nano-magnetic bead immune probe includes a carboxyl nano-magnetic bead and an anti-CEA monoclonal antibody; the chromatographic pad is sequentially provided with a first quality detection line, a second detection line and a quality control line, and the quality control line is more closer to the absorbent pad than the first detection line; the first detection line is provided with a murine NSE detection antibody, and the second detection line is provided with a murine CEA detection antibody; and the quality control line is provided with a goat-anti-mouse second antibody. The immunochromatographic detection test strip can greatly improve the sensitivity and the accuracy of detection of the NSE and the CEA.

Description

technical field [0001] The invention relates to a test strip and a preparation method and an application method thereof, in particular to an immunochromatographic detection test strip for joint detection of NSE and CEA, a preparation method and an application method thereof. Background technique [0002] Near-immune chromatography technology is a new type of detection technology that combines immunolabeling technology and chromatography technology. It can use the chromogenic properties of markers to achieve qualitative, semi-quantitative, and quantitative analysis of the analyte. It uses the microporous filter membrane as the solid phase carrier, the liquid to be tested as the mobile phase, and uses the capillary action and siphon effect of the microporous filter membrane to guide the liquid to be tested to flow forward. The antigen or antibody on the filter membrane reacts to form an immune complex, which stays on the detection line. Through the optical density analysis of ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/574G01N33/58
CPCG01N33/57473G01N33/57488G01N33/587
Inventor 王侃卢文婷崔大祥何井华
Owner SHANGHAI JIAO TONG UNIV
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