Piper D, Nicholson B L, Dunn J
Infect Immun. 1973 Aug;8(2):249-54. doi: 10.1128/iai.8.2.249-254.1973.
Cell cultures of trout gonad tissue (RTG-2) and Atlantic salmon heart, kidney, liver, and spleen tissue were inoculated with 50 50% tissue culture infective doses (TCID(50)) of infectious pancreatic necrosis (IPN) virus per cell, and the titer of cell-associated and released virus was determined from 2 to 16 h postinoculation (PI). Cover slips were collected over the same period and stained for IPN viral antigen by the direct immunofluorescent (FA) technique. Viral replication was detected after a latent period of approximately 2 to 4 h and reached a peak titer of 10(8.2) to 10(8.4) TCID(50) per ml at 8 to 10 h PI. The release of virus was more rapid in Atlantic salmon cells than in RTG-2 cells. Viral antigen was first detected by FA from 3 to 4 h PI. Approximately 75 to 80% of the cells contained antigen in the cytoplasm 9 to 11 h PI. The direct FA technique was found to be a sensitive method for detecting IPN virus in infected cells. Three strains of IPN virus were tested for serological cross reactions by FA and virus neutralization tests.
将50个50%组织培养感染剂量（TCID₅₀）的传染性胰腺坏死（IPN）病毒接种到每个细胞的鳟鱼性腺组织（RTG - 2）以及大西洋鲑鱼的心脏、肾脏、肝脏和脾脏组织的细胞培养物中，并在接种后2至16小时（PI）测定细胞相关病毒和释放病毒的滴度。在同一时期收集盖玻片，并通过直接免疫荧光（FA）技术对IPN病毒抗原进行染色。在大约2至4小时的潜伏期后检测到病毒复制，在接种后8至10小时达到每毫升10⁸·²至10⁸·⁴TCID₅₀的峰值滴度。大西洋鲑鱼细胞中病毒的释放比RTG - 2细胞更快。通过FA在接种后3至4小时首次检测到病毒抗原。在接种后9至11小时，约75%至80%的细胞在细胞质中含有抗原。发现直接FA技术是检测感染细胞中IPN病毒的灵敏方法。通过FA和病毒中和试验对三株IPN病毒进行血清学交叉反应测试。
