ORFV virulence weakening method

Abstract

The invention relates to an ORFV virulence weakening method. A calf testicle cell line is utilized to carry out continuous passage culture of the ORFV, so that the ORFV virulence is weakened, thereby providing a stable and reliable method for preparing and producing an ORFV virulence weakening vaccine; and the method is used for obtaining abundant stable vaccines. The method disclosed by the invention comprises the following steps: 1) separating and initially culturing the ORFV; and 2) carrying out continuous passage culture on the ORFV in the calf testicle cell line until the virulence is weakened. In the step 1), the initial culture of the ORFV adopts the calf testicle cell line, and the culture fluid of the calf testicle cell line is M199, DMEM (Dulbecco Modified Eagle Medium) (high-sugar) and RPMI-1640 culture media. In the step 1), the ORFV is separated from lip scab of a sicken goat. In the step 2), the calf testicle cell line is prepared by introducing a telomerase reverse transcriptase (hTERT).

Description

1. Technical field: [0001] The invention relates to a method for weakening goat aphthus virus toxicity, in particular to a method for continuously subculturing goat aphthus virus by using a calf testis cell line to weaken the toxicity. 2. Background technology: [0002] Ovine contagious impetigo, commonly known as Orf virus, is a zoonotic infectious disease caused by Orf virus in sheep and goats. The World Organization for Animal Health (OIE) lists the disease as a notifiable animal disease , my country lists it as a first-class animal disease. The disease is distributed in almost all sheep-raising countries in the world, and mainly affects sheep and goats under natural conditions, and goats are more frequent. The disease is often popular in groups, and lambs aged 3 to 6 months are most susceptible to infection, and adult sheep are less likely to be infected. Sick sheep and infected sheep are the source of infection of the disease. The virus can be excreted with the saliva,...

AI Technical Summary

Problems solved by technology

However, with the prevalence of sheep oral disease, a large number of local strains have been identified, and many serotypes of sheep mouth disease virus have emerged. The original attenuated strain cannot provide effective protection for all goats.

Moreover, the attenuation of the previous attenuated strains was made by continuously culturing the primary calf testicular cells with the wild strain of sheep mouth ulcers, obtaining cell cultures, adding appropriate protective agents, and processing them. This requires a large number of calf testes to carry out Isolation and culture, time and money consuming

At the same time, when calf testicular cells are used for subculture, the source of calf testis is unstable, which may easily cause batch-to-batch variability

At the same time, the imperfect quarantine of the calf source will cause the pollution of the vaccine production

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