Colloidal gold immunochromatography test strip for detecting bovine rotavirus, and preparation method and application thereof

An immunochromatographic test paper and the technology of bovine rotavirus, applied in the field of medicine, can solve the problems that are not conducive to large-scale detection, unsuitable for on-site detection, and high requirements, and achieve good sensitivity, fast detection speed, and convenient storage.

Active Publication Date: 2018-06-15
NORTHEAST AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, the method is limited and not conducive to large-scale detection
Immunofluorescence technology, enzyme-linked immunosorbent assay (ELISA), nucleic acid electrophoresis, reverse transcription po

Method used

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  • Colloidal gold immunochromatography test strip for detecting bovine rotavirus, and preparation method and application thereof
  • Colloidal gold immunochromatography test strip for detecting bovine rotavirus, and preparation method and application thereof
  • Colloidal gold immunochromatography test strip for detecting bovine rotavirus, and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0085] Example 1 Screening and establishment of hybridoma cells secreting monoclonal antibody against bovine rotavirus VP6 protein

[0086] 1 test material

[0087] 1.1 Strains, cells and viruses

[0088] The recombinant strain pProHTa-NCDV-VP6 / Rosetta expressing bovine rotavirus VP6 protein was constructed and preserved by our laboratory (the recombinant strain has been recorded in the following literature: "Establishment of indirect ELISA detection method for bovine rotavirus VP6 protein", Zhang Yiting et al., Chinese Veterinary Science, 2010, 40(10), 1039-1043); BRV NCDV cell-adapted strains were purchased from China Veterinary Drug Control Institute, and MA104 cells were preserved by our laboratory.

[0089] 1.2 Experimental animals

[0090] SPF grade BALB / c mice and healthy female rabbits weighing 2-3 kg were purchased from Liaoning Changsheng Biotechnology Co., Ltd.

[0091] 1.3 Reagents

[0092] (1) Solution I: 12g urea, 1.56g NaH 2 PO 4 2H 2 O and 0.12g Tris-Bas...

Embodiment 2

[0145] Embodiment 2 is used to detect the preparation of the colloidal gold immunochromatography test strip of bovine rotavirus

[0146] 1 material

[0147] 1.1 Viruses and Cells

[0148] Bovine rotavirus NCDV strains, PoRV, TGEV, PEDV, and BPV are all preserved by our laboratory. MA-104 cells and SP2 / 0 cells were preserved by the laboratory; bovine rotavirus VP6 monoclonal antibody cell 4D-10 strains were prepared by our laboratory.

[0149] 1.2 Experimental animals

[0150] SPF-grade 10-12-week-old BALB / c female mice were purchased from Liaoning Changsheng Biotechnology Co., Ltd.; healthy female rabbits, weighing 2-3 kg.

[0151] 1.3 Immunochromatographic materials

[0152] Nitrocellulose membrane (M135) was purchased from Milipore, USA; glass fiber membrane (GL0194), absorbent paper (H5076), sample pad (GL-b02), PVC board (A-9), connecting tape, and rapid detection plastic card were purchased from Shanghai Jieyi.

[0153] 1.4 Preparation of main reagents

[0154] 1.4...

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Abstract

The invention discloses a colloidal gold immunochromatography test strip for detecting a bovine rotavirus, and a preparation method and an application thereof. The test strip includes a sample pad, acolloidal gold pad, a nitrocellulose membrane and an absorbent filter paper which are sequentially connected from left to right, and also includes a PVC base plate positioned at the bottom, wherein the PVC base plate is used to provide an assembling platform, the nitrocellulose membrane is provided with a quality control line formed by spraying a goat anti-mouse IgG and a detection line formed byspraying a rabbit anti-bovine rotavirus polyclonal antibody, the colloidal gold pad is coated with a colloidal gold particle-labeled mouse anti-bovine rotavirus VP6 protein monoclonal antibody, and the sample pad provides a position for adding a sample to be detected, wherein the mouse anti-bovine rotavirus VP6 protein monoclonal antibody is secreted by a hybridoma cell strain preserved in China General Microbiological Culture Collection Center with the preservation number being CGMCC NO.14726. The test strip prepared in the invention has the advantages of good sensitivity, high specificity, simplicity and rapidness in operation, and suitableness for onsite detection.

Description

technical field [0001] The invention relates to a test strip for detecting bovine rotavirus, in particular to a colloidal gold immunochromatographic test strip for detecting bovine rotavirus prepared based on colloidal gold technology and its preparation method and application. It belongs to the field of medical technology. Background technique [0002] Bovine rotavirus (Bovine Rotaviens, BRV), which belongs to the genus Rotavirus in the Reoviridae family, is one of the main pathogens causing diarrhea in calves. Bovine rotavirus mainly infects 15-45-day-old calves. The early symptoms of the disease are depression, salivation, and diarrhea. If not treated in time, the sick cattle may suffer from dehydration and hypothermia, and are prone to secondary bacterial infection and death. Bring serious harm and huge economic loss to my country's cattle industry. Therefore, the research and development of new technologies for detection of bovine rotavirus, and the establishment of r...

Claims

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Application Information

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IPC IPC(8): G01N33/68
CPCG01N33/68
Inventor 乔薪瑗王丽李一经唐丽杰徐义刚崔文姜艳平
Owner NORTHEAST AGRICULTURAL UNIVERSITY
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