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A kind of nucleic acid aptamer of bovine infectious rhinotracheitis virus and its use

A rhinotracheitis virus and nucleic acid aptamer technology, applied in the field of molecular immunity, can solve the problems of unstable antibody preparation technology, difficult to be suitable for clinical application, and backwardness

Inactive Publication Date: 2019-07-26
BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the different defects in the existing detection methods of IBRV, especially those detection methods based on antibodies, due to the instability of antibodies and the backwardness of preparation technology, these methods are difficult to be suitable for clinical application, and nucleic acid aptamers Superior to antibodies in many ways, stable nucleic acid aptamers may become a substitute for antibodies in future diagnostics

Method used

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  • A kind of nucleic acid aptamer of bovine infectious rhinotracheitis virus and its use
  • A kind of nucleic acid aptamer of bovine infectious rhinotracheitis virus and its use
  • A kind of nucleic acid aptamer of bovine infectious rhinotracheitis virus and its use

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Experimental program
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Embodiment 1

[0031] 1. Test method

[0032] 1. Inoculate IBRV to MDBK cells growing in a single layer at 1% virus dose, and collect the virus when the CPE reaches 80% to 90%. Freeze and thaw repeatedly at -40°C for 3 times, centrifuge at 5000r / min (4500×g) for 30min to remove cell debris, put the supernatant into a treated dialysis bag, and then concentrate the virus solution with PEG6000. Then the concentrated virus solution was ultracentrifuged at 35000r / min (54200×g) for 2h, and finally 1 / 100 of the original cell culture was dissolved and precipitated with 0.01M PBS. After the protein concentration of the diluted product was determined, it was aliquoted and frozen at -20°C until use.

[0033] 2. Purify the concentrated virus liquid obtained above by ultracentrifugation with sucrose density gradient (20%, 40%, 60%), obtain the separated virus liquid after 30,000 rpm and 90 min ultracentrifugation, add an equal volume of STE solution to the The virus sugar mixture was desugared, and the...

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Abstract

The invention provides an infectious bovine rhinotracheitis virus (IBRV) aptamer. The aptamer has the maximum binding force and favorable specificity, can inhibit the virus infection on cells to some extent, and can be used for indirect immunofluorescent assay (IFA) after being marked by FAM. The aptamer can be used for inhibiting the virus infection on cells to perform the antivirus action, and can also be used as an anti-IBRV probe or target spot for preparing anti-IBRV drugs or reagents.

Description

technical field [0001] The invention belongs to the field of molecular immunity, and in particular relates to a nucleic acid aptamer of bovine infectious rhinotracheitis virus and its application. Background technique [0002] Infectious bovine rhinotracheitis (IBR), also known as "red nose" or "necrotizing rhinitis", is an acute, febrile, contact infection of cattle caused by the infectious bovine rhinotracheitis virus (IBRV) sick. The clinical symptoms are divided into 5 types, respiratory tract type, reproductive tract type, conjunctivitis type, abortion type, and encephalitis type. In 1980, my country isolated the virus from cattle imported from New Zealand. Due to improper immune protection measures at that time, the disease was on the rise, which had a great impact on the fattening rate, milk production and reproduction of cattle herds. cause huge economic losses. The disease is listed as a B-type animal infectious disease by the World Organization for Animal Health ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/115G01N33/569G01N33/53A61K48/00A61K31/711
CPCA61K31/711C12N15/115C12N2310/16G01N33/5308G01N33/56983
Inventor 李永清许健张喜喜黄秀芬
Owner BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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