转移贴-甲型H1N1流感病毒小鼠适应株

rojjer

安使
这个病毒不致小鼠死亡，，具体怎么在小鼠上传代，，让他能致死小鼠呢？望高手赐教！！拜谢！！

rojjer

douding
既然是适应株，应该会致死。可能毒力不够。需要鸡胚或鼠肺上活化

安使
这个病毒现在是鸡胚毒，想让它成为小鼠肺脏适应株，具体在小鼠上怎么活化？？谢谢、。。  


Rojjer
mouse-to-mouse intranasal passage
一般传几代就会发现小鼠可以致死，
不过理想的mouse-adapted influenza A virus最好使得小鼠在7天可以全部死亡.
To prepare adapted virus to mice, lung-to-lung passages were performed. Briefly, the BALB/c mice were anesthetized and inoculated with 20 μl of the abovementioned
H5N1 viral suspension by intranasal drip. At the 3rd day after inoculation, the mice were sacrificed, andtheir trachea and lungs were taken out and washed 3 times with a total of 2 ml of phosphate buffered saline (PBS) containing 0.1% bovine serum albumin (BSA). The bronchoalveolar washings were collected and used for infecting the next batch of mice after removing the cellular debris by centrifugation .The lung-to-lung passages
were repeated at least three times and the adapted viruses were harvested, aliquoted, and stored in at -80°C.

reference:A single immunization with HA DNA vaccine by electroporation induces early protection against H5N1 avian influenza virus challenge in mice
http://www.biomedcentral.com/1471-2334/9/17/


douding
简单地说就是在通过小鼠肺悬液反复传代，获得鼠肺适应株。


安使
哦  谢谢楼上啊  现在开始第二次传代  发现第一次滴鼻后小鼠精神不好，抱团，还嗜睡  这第二次观察好像更加活泼了  这是？？谢谢
5# Rojjer 现在小鼠上传代了六次了，还是不致死。。急啊！！要用来做疫苗的保护率实验的  你讲的是禽流感。。这个是猪流感。是不是更不容易做啊？？

lifeng：
产生免疫了？

suenmomo
按照5# Rojjer管理员的方法是可以的
可能楼主在稀释病毒以及滴鼻的时候有些许差异，毒株可能也存在不同差异  


bfvirus
传个10多代才可以。没有那么快，接着传带就可以的。

rojjer

安使：
谢谢各位啦  后来传了六代能致死了 哈哈 实验现在也完成了 谢谢各位