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STLV double-antigen sandwich enzyme linked immunosorbent detecting method and its reagent kit

An enzyme-linked immunosorbent, double-antigen sandwich technology, applied in the field of medicine and biology, can solve the problems of missing or wrong addition of enzyme-labeled antigens and specimens, affecting the accuracy of test results, and decreasing immune activity, so as to reduce the cost of testing and save money. Manpower, lowering effect of lowering activity

Inactive Publication Date: 2008-08-20
GUANGDONG ENTOMOLOGICAL INST
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Problems solved by technology

[0009] In the above ELISA method, due to many steps, omission or wrong addition of the enzyme-labeled antigen and the specimen often occurs, the phenomenon of specimen contamination of the enzyme-labeled antigen, and the enzyme-labeled antigen stored in the solution state may be generated during transportation and storage. Various reasons lead to a decrease in immune activity, which in turn affects the accuracy of the detection results. Therefore, the present invention aims to establish a convenient and feasible double-antigen sandwich rapid detection method to improve the detection sensitivity, speed up the detection speed, and reduce the time required for adding samples. Pollution problems, saving manpower and material resources, and reducing testing costs

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  • STLV double-antigen sandwich enzyme linked immunosorbent detecting method and its reagent kit

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Embodiment 1

[0035] STLV double-antigen sandwich enzyme-linked immunosorbent assay, including the following steps:

[0036] (1) Preparation of enzyme-linked plates

[0037] 1. Antigen coating: Coat 100 μL of STLV-specific antigen with a concentration of 6 μg / mL (0.1 mol / l PBS solution can be used as antigen diluent) to coat the solid phase carrier, so that after the solid phase antigen is formed, the unbound antigen is washed away and impurities, dry in the shade at 4°C;

[0038] 2. Seal with coated plate: (120 μL) place the blocking solution at 4°C for 12 hours, shake and dry in the shade;

[0039] 3. Adding concentration is 8×10 -2 The μg / mL (50 μL) enzyme-labeled STLV antigen was dried in a vacuum freeze-drying oven for 2 hours to allow it to attach to the surface of the solid-phase antigen; after taking it out, let it rest at room temperature for 1 hour to obtain the prepared enzyme-linked plate.

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Abstract

The invention discloses a STLV double-antigen sandwish ELISA and the kit thereof, the test method includes the following steps: 1) STLV coating antigen; 2) coating plate close; 3) STLV with added enzyme label; 4) adding tested sample and the sample diluent, which makes the antibody in sample and the antigen complex on the solid phase carrier react completely, solid antigen-antibody-enzyme label antigen complex is formed, washing off the uncombined antibody and impurity; 5) adding chromogenic substrate, through color comparisons, measuring the amount of antibody. The improved STLV double-antigen sandwish ELISA and the kit thereof have advantages of fast test speed, human and material resources saving and reduced test cost. The method is fast, simple and clear, easy to be popularized, and meets the basic needs, so the method is suitable for epidemiological study.

Description

technical field [0001] The invention belongs to the field of medical biology, and specifically relates to an improved STLV double-antigen sandwich enzyme-linked immunosorbent (ELISA) assay method and a kit thereof. Background technique [0002] Simian T-lymphotropic virus type 1 (STLV-1) is an important infectious virus in monkeys. Functional disorder, thus affecting the research of animal experiments. As early as 1992, the "Standards for Medical Experimental Animals" promulgated by the Ministry of Health of the People's Republic of China stipulated that STLV-1 type is one of the viruses that SPF experimental monkeys must exclude. With the increasing demand for SPF experimental monkeys in the current international market, STLV-1 virus has been listed as one of the pathogens that must be excluded. [0003] Enzyme-linked immunosorbent assay (ELISA) is an enzyme immunoassay technology developed after immunofluorescence and radioimmunoassay techniques, involving many sciences ...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N33/535
Inventor 季芳万玉玲饶军华刘晓明
Owner GUANGDONG ENTOMOLOGICAL INST
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