Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Subgroup J avian leukosis antibody quick test paper card, and application thereof

A technique for detecting avian leukaemia and test strips, which is applied in the direction of measuring devices, instruments, scientific instruments, etc. It can solve the problems of high technical level of testing personnel, unsuitability for rapid clinical detection, complicated operation, etc., and achieve rapid response, simple operation, and high sensitivity. high effect

Active Publication Date: 2012-06-20
INST OF ANIMAL SCI & VETERINARY HUBEI ACADEMY OF AGRI SCI
View PDF5 Cites 3 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The PCR method has high requirements on the technical level of testing personnel and equipment, high cost, complicated operation, prone to false positives and false negatives, and is not suitable for rapid clinical detection

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Subgroup J avian leukosis antibody quick test paper card, and application thereof
  • Subgroup J avian leukosis antibody quick test paper card, and application thereof
  • Subgroup J avian leukosis antibody quick test paper card, and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1 Preparation of mouse anti-chicken IgG Fc fragment monoclonal antibody

[0027] Collect the hyperimmune serum of laying hens before laying in large-scale laying hen farms and purify by caprylic acid-ammonium sulfate method, add 4 parts of sodium acetate buffer solution for each part of serum: concentration 0.06mol / L, pH value 4.0); Adjust the pH value to 4.5 with 1mol / L NaOH, and add octanoic acid 33μl / ml. Add while stirring for 30min. Centrifuge at 12,000 rpm for 10 min at 2-8°C and discard the precipitate. After filtering the supernatant with filter paper, adjust the pH value to 7.4 with NaOH, then slowly add saturated ammonium sulfate to a final concentration of 45%, continue to stir for 30 minutes, and stand at 2-8°C for 2 hours. Centrifuge at 12,000 rpm for 30 min at 2-8°C and discard the supernatant. Dialyze against 0.01M tTris (pH 9.0) to desalt until desalted with BaCl 2Concentrate with PEG8000 until no white precipitate is detected. The crudely ...

Embodiment 2

[0029] Example 2 Preparation of J Subgroup Avian Leukemia Subgroup Specific gp85 Antigen Protein

[0030] Genetic engineering technology is used to highly express the J subgroup avian leukemia subgroup specific gp85 antigen protein. Referring to the gp85 gene sequence of the J subgroup prototype strain HPRS-103 that has been published on GENBANK, a pair of primers for amplifying the gp85 gene was designed. TG-3'. EcoRI and XhoI restriction sites were introduced into the 5' ends of P1 and P2, respectively. The amplification length is 921bP, and the reaction conditions are: 95°C pre-denaturation for 5 minutes, followed by 33 cycles of 94°C for 30s, 57°C for 45s, 72°C for 1min, and finally 72°C for 10min, and the PCR product was connected to the pMD-18T vector. Transform the ligation product into DH5α-competent medium, smear it on an LB plate containing 100g / mL ampicillin (Amp), incubate at 37°C for 16 hours, pick a single colony, extract the recombinant plasmid by alkaline l...

Embodiment 3

[0032] Example 3 Preparation of rabbit anti-mouse IgG polyclonal antibody

[0033] The blood of hyperimmunized mice was collected, the serum was separated, and the mouse serum IgG was crudely extracted by the octanoic acid-ammonium sulfate method, and purified by Superdex200 gel chromatography (same as Example 1). The protein content of mouse IgG was measured to be 13.1 mg / mL, can be used for the preparation of rabbit anti-mouse IgG polyclonal antibody.

[0034] Fully emulsify with mouse IgG protein of 50-100ug / kg body weight plus Freund's adjuvant, immunize healthy New Zealand white rabbits by subcutaneous and intramuscular injection 3-4 times, 10 days after the last immunization, blood is collected from vein, and the serum antibody effect is determined by ELISA When the titer was above 1:2000, the hyperimmune serum was collected by collecting blood from the heart or carotid artery; the IgG from immune rabbit serum was crudely extracted by the octanoic acid-ammonium sulfat...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

PropertyMeasurementUnit
concentrationaaaaaaaaaa
diameteraaaaaaaaaa
Login to View More

Abstract

The invention relates to a subgroup J avian leukosis antibody quick test paper card. The test paper card comprises a substrate; a sample pad, a gold mark pad, a nitrocellulose membrane and a water absorption pad are all arranged on the substrate; the gold mark pad is coated with a mouse anti-chicken immunoglobulin G (IgG) Fc fragment monoclonal antibody which is marked by colloidal gold; and the nitrocellulose membrane is coated with a detection line consisting of avian leucosis virus-J subgroup specificity gp85 antigenic protein and a quality control line consisting of rabbit anti-mouse IgG. The invention has the advantages that: by a method for quickly detecting the subgroup J avian leukosis antibody by using the quick test paper card, the subgroup J avian leukosis antibody in chicken serum can be quickly detected, so that whether chicken is infected with subgroup J avian leukosis or not is judged. The test paper card has the advantages of high reaction speed and sensitivity, strong specificity, convenience in operation, and economy and practicality and is suitable for 'animal yard side' detection.

Description

technical field [0001] The invention relates to a test paper card for rapid detection of J subgroup avian leukemia antibody, which can retrospectively determine whether chickens have been infected with J subgroup avian leukemia virus by detecting the J subgroup avian leukemia antibody. Background technique [0002] Subgroup J avian leukemia is a neoplastic infectious disease caused by subgroup J avian leukemia virus (ALV-J) infection. ALV-J is an enveloped retrovirus. Clinical infection of poultry mainly manifests as myelocytoma, immune tolerance, high mortality and growth retardation. From 1997 to 1998, ALV-J broke out globally, causing a devastating blow to the world broiler breeder industry. In addition to myeloma caused by ALV-J infection, erythroblastoma, hemangioma, nephroma, and sarcoma often occur in the late stage of infection. The mortality rate is usually 1% to 5%, and the peak period can reach 50%. Studies have confirmed that ALV-J has a lasting damage to the i...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577
Inventor 罗青平张蓉蓉温国元邵华斌艾地云王红琳杨峻罗玲刘丽娜
Owner INST OF ANIMAL SCI & VETERINARY HUBEI ACADEMY OF AGRI SCI
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com