Quantum dot immunochromatography test strip for synchronous and quantitative joint inspection of HBsAg, HBeAg and HBcAb and method for synchronous and quantitative joint inspection of HBsAg, HBeAg and HBcAb
A technology for joint detection of hepatitis B and immunochromatography, applied in measuring devices, analytical materials, biological testing, etc.
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Embodiment 1
[0041] Example 1: Quantum dot immunochromatography test strip structure for simultaneous quantitative joint detection of HBsAg, HBeAg, and HBcAb
[0042] combine figure 1 be explained.figure 1 Among them, the quantum dot immunochromatographic test strip for the simultaneous quantitative joint detection of the three items of hepatitis B includes a sample pad 1, a red blood cell filter membrane 2, a marker pad 3, an analysis membrane 7, and Absorbent pad6.
[0043] The marking pad 3 of the test strip is a glass fiber membrane. The analysis membrane 7 is a nitrocellulose membrane, a nylon membrane, or a nitrocellulose / acetate mixed membrane, on which there are a detection zone (ie T zone) 4 and a quality control zone (ie C zone) 5 . Described backing 8 is polyester or plastic plate.
[0044] The marker pad 3 is coated with CdSe / ZnS QD 526 Labeled mouse anti-HBsAg monoclonal antibody (Mouse anti-HBsAg McAb -CdSe / ZnS QDs 526 ), CdSe / ZnS QDs 615 Labeled mouse anti-HBeAg monoc...
Embodiment 2
[0045] Example 2: The preparation method of the quantum dot immunochromatography test strip for simultaneous quantitative joint detection of HBsAg, HBeAg, and HBcAb
[0046] The preparation method of described test strip comprises the steps:
[0047] A. Quantum dot conjugated antibody / antigen:
[0048] a) Take water-soluble quantum dots CdSe / ZnS QD with emission wavelengths of 526nm, 615nm, and 566nm 526 、CdSe / ZnS QD 615 、CdSe / ZnS QD 566 Use PBS buffer to adjust pH=6-9, respectively add EDC (l-(3-dimethylaminopropyl)-3-ethylcarbodiamine hydrochloride) and NHS (N-hydroxythiosuccinic acid imine) activated at room temperature for 10-60min;
[0049] b) Mouse anti-HBsAg monoclonal antibody (Mouse anti-HBsAg McAb ), mouse anti-HBeAg monoclonal antibody (Mouse anti-HBeAg McAb ), HBcAg vortex shaking reaction for 0.5-3h;
[0050] c) Add bovine serum albumin (BSA) respectively, and block the reaction in the dark for 0.5-2h;
[0051] d) Each product was purified by centr...
Embodiment 3
[0061] Example 3: Preparation method of the standard curve for the simultaneous quantitative joint detection of HBsAg, HBeAg, and HBcAb by using the quantum dot immunochromatographic test strip for simultaneous quantitative joint detection of hepatitis B three items
[0062] (a) Take HBsAg, HBeAg, HBcAb standard products respectively with phosphate buffered solution (PBS) to make several parts of standard product serial concentration in the way of doubling dilution.
[0063] (b) Add the concentration of each standard substance dropwise on 10 quantum dot immunochromatography test strips and detect with a detector under the same conditions (that is, use 10 quantum dot immunochromatography test strips for each standard substance concentration on the same 10 times with the detector under the conditions), and read the fluorescence intensity of the T band (OD t ) vs. C-band fluorescence intensity (OD c ), to get the mean and OD t / OD c ratio.
[0064] (c) Take the concentrati...
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