Imunofluorescencia realizada em cerebros de camundongos infectados com virus rabico-cepa CVS, em diferentes estagios de decomposicao / Immunofluorescence performed in brain of rabies virus infected mice - CVS cepa, at different stages of decomposition

O teste de imunofluorescencia (IF) foi avaliado na deteccao de virus rabico presente em cerebros de carcacas de camundongos infectados com virus da cepa CVS, os quais foram conseguidos atraves de uma combinacao de tratamentos, em que se variaram as temperaturas (4,25 e -20 graus Celsius) e o tempo de armazenamento. No teste de IF realizado com impressoes cerebrais de carcacas que haviam sido submetidas a temperatura de 25 graus Celsius por 12-18h, houve maior dificuldade de visualizacao imediata dos corpusculos de inclusao, enquanto que nos materiais conservados a 4 graus Celsius por ate 48h, as inclusoes foram facilmente reconhecidas. Carcacas mantidas a -20 graus Celsius mantiveram-se viaveis a identificacao pela IF mesmo apos terem sido armazenadas por 720h quando foram feitas as ultimas observacoes. Em carcacas mantidas a 25 graus Celsius por 10h, com tratamento posterior a 4 e -20 graus Celsius, o antigeno rabico nao pode ser identificado atraves da IF, em consequencia da decomposicao das carcacas que ocorrem, respectivamente, apos 10 e 24h. Recomenda-se, portanto, empregar o teste de IF, em carater de rotina, no controle de qualidade da vacina contra a Raiva, no que diz respeito a prova de virus residual (teste de verificacao da inativacao viral), de vez que ele permite esclarecer mortes assintomaticas...

[Immunofluorescence performed in brain of mice, infected with the CVS strain of the rabies virus, in different stages of decomposition]. / Imunofluorescência realizada em cérebros de camundongos infectados com vírus rábico--cepa CVS, em diferentes estágios de decomposição.

The efficiency of the fluorescent antibody (FA) test in detecting rabies virus antigen in decomposed specimens was evaluated in simulated conditions of the safety test recommended for the assessment of residual virus in inactivated rabies vaccines. The CVS-infected mice were submitted to different treatments combining time and temperature in order to cause different stages of carcass decomposition and, the FA test was carried out sequentially at pre-determined time intervals. For the materials stored at 25 degrees C, greater difficulties for prompt recognition of the inclusion bodies were found after 12-18h, whilst the specimens maintained at 4 degrees C, the inclusions were easily visualized for up to 48h. Brain smears of carcasses kept at -20 degrees C were suitable for adequate identification after 720 h of storage. In carcasses that had been maintained at 25 degrees C for 10 h with additional storage at 4 or -20 degrees C, rabies antigenicity could not be detected, respectively after 10 and 24 h, due to tissue decomposition. The authors recommend that the FA test, when applied as an additional tool for the control of the safety test of inactivated rabies vaccine using mice, care must be taken in order to avoid the use of decomposed materials.