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Respiratory syncytial virus detection kit

A detection kit and technology for syncytial virus, applied in the field of enzyme immunoassay, can solve the problems of difficult interpretation, long time, high cost, sometimes even weeks to produce results, and inappropriateness.

Inactive Publication Date: 2015-11-25
TIANJIN BAODI HOSPITAL
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Problems solved by technology

Indirect immunofluorescence method is commonly used in clinical laboratories to detect RSV, followed by virus isolation and culture method, PCR method, virus cell culture takes a long time, high cost, sometimes even a few weeks to get the results, not suitable for clinical testing, indirect immunofluorescence method The operation is simple and convenient, and it is suitable for the detection of RSV in clinical laboratories. The kits represented by the German Ommon Company are the most widely used. For the reaction, goat anti-human IgM labeled with fluorescein isothiocyanate was added, and donkey anti-human IgG antibody was required to absorb anti-RSV IgG antibody before the experiment. The biggest disadvantage of this method is the non-specific binding of IgG antibody in the body, resulting in false Positive, difficult to interpret results

Method used

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Embodiment Construction

[0008] 1. Preparation of Respiratory Syncytial Virus ELISA Plate

[0009] Dilute the anti-respiratory syncytial virus antibody to 1:200 times with the coating solution, add 100 μl to the microwells of the coated microwell reaction plate, add 350 μl / well of phosphate buffer containing 1% bovine serum albumin, and seal at room temperature 2 hours, overnight at 4°C. After shaking off the liquid in the hole, wash 3 times with washing solution, 3 minutes each time.

[0010] 2. Preparation of biotinylated antibody

[0011] Preparation of biotinylated antibody with 0.1mmol / L NaHCO 3 Dilute the goat anti-human IgM respiratory syncytial virus antibody to 1mg / L, add biotin N-hydroxysuccinimide ester dissolved in dimethylformamide to each milliliter of antibody solution, add NaHB 4 Mix 0.1ml, and dialyze against 0.01mol / L pH7.4 phosphate buffer at 4°C for 2 hours, overnight at 4°C. After adding an appropriate amount of neutral glycerin, aliquot it in small quantities.

[0012] 3. Pr...

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Abstract

The invention discloses a respiratory syncytial virus kit and a detection method thereof, belonging to the technical field of enzyme immunoassay. According to the respiratory syncytial virus kit, the pharyngeal of a child is wiped by adopting a throat swab, nasopharyngeal secretions of the throat swab are dissolved by an ambroxol hydrochloride medicine, nasopharyngeal epithelial cells as sediments are added in 2.0ml of a phosphate buffer solution containing 1.0 percent (V / V)Triton X-100 after being centrifuged at a high speed, stand for 30min, and are violently oscillated so that respiratory syncytial viruses fall off from the nasopharyngeal epithelial cells, a dissolved solution is taken to be added on a respiratory syncytial virus detection ELISA (Enzyme Linked Immunosorbent Assay) plate, the respiratory syncytial viruses are detected by adopting an ELISA method, warm bath and washing are carried out, a biotinylated respiratory syncytial virus antibody is added, streptavidin is coupled with a streptavidinbiotin peroxidase complex, and a substrate liquid is added, thus a result is read. The detection method is simple to operate, is rapid, sensitive and accurate, and is low in cost, is suitable for detecting the infection of nasopharyngeal epithelial cells by the respiratory syncytial viruses, and can be used as an important inspection index of infecting the respiratory syncytial viruses by the child.

Description

technical field [0001] A respiratory syncytial virus detection kit and a detection method thereof belong to the technical field of enzyme immunoassay. Background technique [0002] At present, domestic detection methods for respiratory syncytial virus (RSV) infection include electron microscope detection, culture identification method, polymerase chain reaction (PCR), enzyme-linked immunosorbent assay, agglutination method, fluorescent immunospecific antibody detection method, etc. In this method, antigen detection is more sensitive than antibody detection. Indirect immunofluorescence method is commonly used in clinical laboratories to detect RSV, followed by virus isolation and culture method, PCR method, virus cell culture takes a long time, high cost, sometimes even a few weeks to get the results, not suitable for clinical testing, indirect immunofluorescence method The operation is simple and convenient, and it is suitable for the detection of RSV in clinical laboratori...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/569G01N33/535
CPCG01N33/56983G01N2333/135
Inventor 徐向英李立和齐金龙杨帅
Owner TIANJIN BAODI HOSPITAL
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