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.