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Method for detecting neutralizing antibodies aiming at avian influenza virus in serum of animals immune to avian influenza vaccine

A technology for avian influenza virus and animal immunization, applied in measuring devices, instruments, scientific instruments, etc., can solve problems such as long time required, difficult to detect in large quantities at one time, difficult to commercialize, etc., and achieve easy standardization, stable and reliable methods, and expanded scope Effect

Inactive Publication Date: 2009-11-25
MICROBE EPIDEMIC DISEASE INST OF PLA MILITARY MEDICAL ACAD OF SCI
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AI Technical Summary

Problems solved by technology

The traditional detection of avian influenza virus neutralizing antibody mainly adopts the whole cell culture method. This method judges the result by observing the cytopathic pathology (CPE), which is highly subjective; the cytopathic pathological change (CPE) usually needs to be observed continuously for 5-7 days to finally confirm the result. It takes a long time; and the detection of highly pathogenic avian influenza antibodies must be carried out in a BSL-3 laboratory; the amount of specimens for one detection is small, and it is difficult to detect a large number of them at one time; this method is difficult to commercialize

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  • Method for detecting neutralizing antibodies aiming at avian influenza virus in serum of animals immune to avian influenza vaccine
  • Method for detecting neutralizing antibodies aiming at avian influenza virus in serum of animals immune to avian influenza vaccine

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Embodiment

[0045] Embodiment, detect the neutralizing antibody against avian influenza virus in the sera of avian influenza vaccine immunized animals

[0046] 1. Neutralizing antibody detection

[0047] (1) Determination of virus titer (tissue culture half infectious dose-TCID 50 Titration)

[0048] 1. Prepare virus dilution

[0049] The virus dilution was prepared as follows:

[0050] RPMI1640+1% bovine serum albumin+antibiotics, ready to use.

[0051] 429ml RPMI1640

[0052] 66 ml 7.5% Bovine Serum Albumin (BSA)

[0053] 5ml penicillin and streptomycin mixture (10000 units / ml)

[0054] 2. Virus dilution

[0055] 1) Take 1 tube of avian influenza virus VNH5N1-PR8 / CDC-rg (US CDC, Safety and immunogenicity of an inactivated subvirion influenza A(H5N1) vaccine.N Engl J Med2006; 354:1343-51.), dilute 1:100 ( 100 microliters of virus solution + 9.9 milliliters of diluent).

[0056] 2) Do serial semi-logarithmic dilution of the 1:100 diluted virus solution to make it 10% of the origi...

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Abstract

The invention discloses a method for detecting neutralizing antibodies aiming at avian influenza virus in serum of animals immune to avian influenza vaccine. The method provided by the invention comprises the steps: mixing inactivated to-be-detected serum with avian influenza virus; co-culturing obtained virus-serum mixture and a host cell of the avian influenza virus; taking an antibody resisting avian influenza virus NP protein as a first antibody to perform enzyme-linked immune response; and determining the titer of a neutralizing antibody of avian influenza virus serum in the to-be-detected serum. The method can be used to detect the effectiveness of the avian influenza vaccine. The method provided by the invention has the advantages of strong objectivity, convenience for standardization, short required time, capability of being performed in BSL-2 laboratories and performing high-throughput detection, objective results, stability and reliability.

Description

technical field [0001] The invention relates to a method for detecting neutralizing antibodies against bird flu virus in serum of animals immunized with bird flu vaccine. Background technique [0002] Avian influenza (avian influenza, AI) is an infection or disease syndrome of poultry and wild birds caused by avian influenza virus (AIV) of the Orthomyxoviridae family Influenzavirus A, including chickens, turkeys, ducks, geese and quails, etc. Poultry, wild birds, waterfowl and seabirds can be infected. This highly contagious infectious disease can present asymptomatic infection, varying degrees of respiratory symptoms, decreased egg production rate, and even cause purplish-black crest and beard, dyspnea, diarrhea, glandular papilla and gizzard subcornea and other organs Extensive tissue hemorrhage, pancreatic necrosis, fibrinous peritonitis, and even acute sepsis with a 100% fatality rate. Wild birds or migratory birds are the natural hosts of AIV. It has been confirmed th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N33/577
Inventor 曹务春张泮河贾娜
Owner MICROBE EPIDEMIC DISEASE INST OF PLA MILITARY MEDICAL ACAD OF SCI
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