Preparation method of standard sample of H9N2 subtype avian influenza virus

Abstract

The invention discloses a preparation method of a standard sample of a H9N2 subtype avian influenza virus. The preparation method comprises the following steps: continuously diluting the H9N2 subtype avian influenza virus with normal saline ten times to the concentration of 10<8>, inoculating five 10-days-old SPF (specific pathogen free) embryonated eggs into each dilution of H9N2 subtype avian influenza virus, wherein each embryonated egg is 0.1ml, continuously hatching the embryonated eggs at the temperature of 37 DEG C after the inoculation, and lighting the embryonated eggs everyday; b, gaining inoculated embryonated eggs dying after 72-120 hours and allantoic fluid of the embryonated eggs living for 120 hours one by one, respectively measuring HA(hyaluronic acid) value of the allantoic fluid of each embryonated egg, and taking the allantoic fluid in one embryonated egg infected by a highly diluted virus; C. orderly repeating the step a and the step b twice, wherein the H9N2 subtype avian influenza virus of the step a is a transferred inoculant produced by the step b in the repetitive process; and d. finally diluting the transferred inoculant produced by the step b with normal saline ten times, inoculating ten-times diluted solution to the inside of an allantoic cavities of the 10 to 11-days-old SPF embryonated eggs, continuously hatching the SPF embryonated eggs, lighting the embryonated eggs everyday to obtain inoculated embryonated egg allantoic fluid dying after 72 to 120 hours, subpackaging resultant with ampoule, and storing the ampoules at the temperature of minus 80 DEG C.

Description

technical field [0001] The invention relates to a preparation method of a virus standard sample, in particular to a preparation method of a H9N2 subtype avian influenza virus standard sample which saves time and effort, has high stability and good uniformity. Background technique [0002] Avian Influenza (AI), also known as true fowl plague or European fowl plague, is a poultry infection and / or disease syndrome caused by type A influenza virus. Influenza virus (Avian Influenza virus, AIV) can be divided into non-pathogenic, low pathogenic (Low Pathogenic Avian Influenza, LPAI) and highly pathogenic (Highly Pathogenic Avian Influenza, HPAI) according to the degree of pathogenicity to poultry three kinds. Symptoms of animals infected with avian influenza virus vary. Non-pathogenic avian influenza has no obvious symptoms; infection with low-pathogenic avian influenza may have mild respiratory symptoms, and sporadic death of sick birds; infection with highly pathogenic avian in...

AI Technical Summary

Problems solved by technology

Moreover, new research shows that the pathogenicity of some H9N2 subtype epidemic strains varies significantly, especially for broiler chickens, the mortality rate can reach more than 30%; difficulty

[0005] At present, people have carried out extensive research on the H9N2 subtype of avian influenza, and the quality control of the laboratory has always been of concern to people. For internal quality control, the existing laboratories basically use the H9N2 subtype of avian influenza prepared by themselves. Samples, not only the sample preparation process is time-consuming and laborious, but also it is difficult to maintain the stability of live bacteria, and it is difficult to compare the results of different laboratories

Images