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Preparation method of NSE test strip, test strip, test card and NSE test kit

A technology for detecting test paper and test strips, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., to achieve the effects of shortened detection time, wide application range, and improved detection efficiency

Pending Publication Date: 2019-03-08
河南省生物工程技术研究中心
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, for NSE, related technologies and detection tools for its quantitative detection by time-resolved fluorescent immunoassay have not been reported yet.

Method used

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  • Preparation method of NSE test strip, test strip, test card and NSE test kit
  • Preparation method of NSE test strip, test strip, test card and NSE test kit
  • Preparation method of NSE test strip, test strip, test card and NSE test kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Embodiment 1: Preparation of NSE detection test strip, detection card and kit set

[0030] 1.1 Preparation of conjugation pads: (1) Preparation of NSE antibody-fluorescent microsphere complex: take 10 μL of Eu-carboxyl fluorescent microspheres (suspension, particle size 200 nm, concentration 112 ueq / g) and add 400 μL of 0.05M boric acid buffer at pH=7.6 solution, mix well; add 40 μL 1mg / ml EDC activator and 10 μL 1mg / ml NHS solution, shake and activate on a constant temperature oscillator for 20 minutes; then centrifuge at 5000 rpm for 20 minutes, discard the supernatant, and add 500 μL 0.05M pH=7.6 boric acid buffer Reconstitute the precipitate in the solution, vortex to mix, add 40ugNSE antibody, vortex to mix, place in a shaker for low-speed oscillating coupling for 2h; centrifuge at 15000rpm for 15min, discard the supernatant, add 1500μL of reconstitution solution to redissolve, and then use 10μL of blocking solution ( 20%BSA) for blocking 30min, the complex solutio...

Embodiment 2

[0051] Embodiment two, control experiment

[0052] 1. The rest of the conditions remain the same, replace the complex solution with 0.01M Tris-Hcl PH = 8.0, 0.1% Tween-20, 1% BSA, 0.1% PVP, 0.05% PEG-200, 0.5% glycine and 3% trehalose solution (the concentration of each of the above components is the final concentration);

[0053] The treatment solution was replaced with: 0.01M Tris-Hcl PH=8.0, 0.1% T-20, 0.1% BSA and 5% sucrose (the concentrations of the above components are the final concentrations);

[0054] Others were all prepared test strips according to the method of Example 1, and further made into test cards, and carried out sample determination.

[0055] 2. Draw the standard curve: (1) Sample acquisition: The blood sample is provided by the hospital and the quality control value is determined. On this basis, the normal saline is used for gradient dilution. The assigned units are: 0, 0.39, 0.78, 1.56 , 3.125, 6.25, 12.5, 25, 50, 100, 200, 300, 400, 500; the real value...

Embodiment 3

[0061] Embodiment three, the detection effect verification of the test strip / detection card that embodiment one makes:

[0062] 1. measure the detection limit of the detection card / test strip qualitative detection that the present invention (embodiment one) makes

[0063] (1) Sample processing: collect blood with the sample collection device provided in this kit.

[0064] (2) Preparation of test card or test strip: place the prepared test card / test strip at room temperature for 15 minutes before use.

[0065] Dilute the NSE standard to 2ng / mL, 4ng / mL, 8ng / mL, 16ng / mL, 32ng / mL, 64ng / mL, 128ng / mL, then add sample saline to the standard for dilution (per drop of sample Add 3mL to simulate the actual detection situation), after dilution, add 65μL to the sample pad for each detection for detection.

[0066] After adding the sample for 5 minutes, read under the ultraviolet light. The results are shown in Table 3:

[0067] Table 3 NSE qualitative detection detection limit determi...

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Abstract

The invention relates to the technical field of immunochromatography, and particularly relates to a preparation method of an NSE test strip, and a test strip, a test card and an NSE test kit preparedaccording to the method. The preparation method of the NSE test strip comprises the following steps: arranging a sample pad, a bonding pad, a nitrocellulose membrane and an absorbent pad on a bottom plate in sequence, wherein an NSE antibody-fluorescent microsphere composite is fixed on the bonding pad, a detection line T and a quality control line C are arranged on the nitrocellulose membrane, the control line C is arranged at the end close to the absorbent pad, the detection line T is coated with an anti-NSE monoclonal antibody, and the control line C is coated with a goat anti-mouse IGg polyclonal antibody. The NSE test strip is used for detecting the NSE test speed without relying on large-scale medical equipment.

Description

technical field [0001] The invention belongs to the technical field of immunochromatography detection, and in particular relates to a preparation method of an NSE detection test strip, a test strip, a detection card and an NSE detection kit. Background technique [0002] Small cell lung cancer (SCLC) is a special type of lung cancer and belongs to highly malignant neuroendocrine tumors. SCLC has the characteristics of short doubling time, high proliferation index and early spread. Small cell lung cancer grows rapidly and tends to appear in the large airways, irritating the bronchial mucosa and obstructing the airways. The most common clinical symptom is irritating dry cough. The first symptoms include cough, sputum, fever, chest tightness, shortness of breath, chest pain and extrapulmonary symptoms. The respiratory symptoms and signs of central lung cancer are earlier and more obvious than those of peripheral lung cancer. Except for chest pain when the mediastinum, pleura or...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/558
CPCG01N33/558G01N33/577
Inventor 王云龙支开旗李玉林王继创张怡青
Owner 河南省生物工程技术研究中心
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