VIb subtype pigeon Newcastle disease positive serum standard substance and preparation method thereof

A technology for positive serum and standard substance, which can be used in material testing products, measuring devices, instruments, etc., and can solve the problems of low antigen homology, long genetic distance, and low homology.

Active Publication Date: 2021-09-10
ZHONGCHONG XINNUO BIOTECH TAIZHOU CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] In recent years, the genetic distance between pigeon Newcastle disease strains isolated from pigeon populations and the conventional chicken Newcastle disease vaccine strain La Sota strain is relatively long, and their nucleotide and amino acid The homology with the Newcastle disease vaccine strain La Sota is relatively low, only 82% to 84%. In the cross-virus neutralization test, the antigen homology between the gene type Ⅵ pigeon strain and the La Sota strain is low to about 65%

Method used

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  • VIb subtype pigeon Newcastle disease positive serum standard substance and preparation method thereof
  • VIb subtype pigeon Newcastle disease positive serum standard substance and preparation method thereof
  • VIb subtype pigeon Newcastle disease positive serum standard substance and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Embodiment 1——Preparation of Pigeon Newcastle Disease Inactivated Vaccine

[0042] 1.1 The preparation of the antigen solution uses sterile saline for pigeon Newcastle disease virus LY / 2020 / Pi strain seed 10 -4 Dilute, inoculate 10-day-old SPF embryos into the allantoic cavity, 0.1 mL per embryo, continue incubation at 37°C, discard chicken embryos that died before 48 hours. At 96 hours, the chicken embryos were taken out and frozen, the allantoic fluid was harvested aseptically, and the hemagglutination value and virus content were determined by sampling. At the same time, the sterility, mycoplasma and exogenous virus tests were carried out according to the appendix of the 2015 edition of "Chinese Veterinary Pharmacopoeia". Add β-propiolactone to the qualified embryo fluid at 1 / 4000 (V / V), mix thoroughly, inactivate at 4°C for 20 hours, hydrolyze at 37°C for 2 hours, and store at 2-8°C. At the same time, samples were taken for sterility test and inactivation test. Th...

Embodiment 2

[0051] Embodiment 2 - the screening of negative pigeon

[0052] 1. Primary screening. Select a pigeon farm in Jiangyin, Rugao and Gaogang, Taizhou for investigation, check the breeding and management conditions of the pigeon farm, ask about the history of disease, vaccination history and veterinary drug use of the pigeon farm. Exclude pigeons with poor feeding and management conditions and that cannot meet the following conditions: 1. Normal food intake and drinking water, shiny feathers, strong vigilance, spiral-shaped feces, and a little white urea; 2. No bacteria, viruses, and parasites and other disease history. After investigation, the breeding and management conditions of the pigeon farms in the three places are relatively good, and no bacterial, viral and parasitic infections have occurred within a year. One pigeon farm with normal spirit, food intake, drinking water, feces, growth and development, etc. was selected from each of the three pigeon farms. Pigeon flocks s...

Embodiment 3

[0058] Embodiment 3——Preparation of Pigeon Newcastle Disease Negative Serum

[0059] The pigeons numbered 397 and 398 screened in Example 2 were aseptically collected blood via the subwing vein. After collection, they were placed in a water bath at 37°C for 30 minutes, centrifuged at 5000r / min for 10min, and the serum was collected. After purification, Use a 0.22μm disposable filter to filter and sterilize, store in 1.0mL / bottle aliquots, and use it as negative serum for HI test and sterility, mycoplasma and exogenous virus tests. The results showed that the HI test of the prepared negative serum was negative, and the purity was good, without bacteria, mold, mycoplasma and exogenous virus contamination. The results of the purity test are shown in Table 4.

[0060] Table 4 Pigeon Newcastle disease positive serum and negative serum purity test results

[0061]

[0062] Note: "-" means no microbial growth or negative.

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Abstract

The invention relates to a preparation method of a VIb subtype pigeon Newcastle disease positive serum standard substance. The HI titer of the positive serum standard substance prepared by the method is not lower than 1:128, the neutralizing titer is not lower than 1:1600, the serum can completely neutralize the VIb subtype pigeon Newcastle disease virus, the positive serum standard substance is used for detecting the specificity of a positive control standard substance and a vaccine seed virus in a hemagglutination inhibition test and an exogenous virus, the accuracy of an antibody detection result can be ensured in the hemagglutination inhibition test, the antibody level of the immunized pigeons is fully reflected, and a guarantee is provided for accurate monitoring of VIb subtype Newcastle disease epidemic strains in pigeons, immunogenicity and specificity detection of pigeon Newcastle disease seed viruses, exogenous virus detection and vaccine immune efficacy evaluation. The invention fills the blank that no standard positive serum exists in the development of VIb subtype pigeon Newcastle disease vaccines and epidemiological investigation. The preparation method of the positive serum comprises the following steps: preparation of a pigeon Newcastle disease LY/2020/Pi strain inactivated vaccine, animal screening, animal immunization, and preparation, purification and inspection of the positive serum.

Description

technical field [0001] The invention relates to a pigeon Newcastle disease subtype VIb positive serum standard product and a preparation method thereof, belonging to the technical field of veterinary biological products. Background technique [0002] Pigeon Newcastle disease is caused by pigeon paramyxovirus type Ⅰ (pigeon paramyxovirus-1, PPMV-1), a highly contagious infectious disease prevalent among pigeons, and its clinical symptoms are similar to chicken Newcastle disease. The epidemiological feature of the disease is that the new epidemic area spreads rapidly, the incidence rate is 100%, and the mortality rate is more than 80%. In recent years, with the rapid development of my country's pigeon industry, the incidence of the disease is on the rise, and even some pigeon farms are outbreaks, which bring greater economic losses to the pigeon industry. Epidemiological surveys in recent years have shown that the Newcastle disease virus prevailing in pigeon populations in my...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/96G01N33/569
CPCG01N33/96G01N33/56983G01N2333/125
Inventor 刁小龙刘云迎何海蓉印广浩栾璐田志伟
Owner ZHONGCHONG XINNUO BIOTECH TAIZHOU CO LTD
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