Tapay L M, Lu Y, Gose R B, Nadala E C, Brock J A, Loh P C
Department of Microbiology, University of Hawaii-Manoa, Honolulu 96822, USA.
J Virol Methods. 1997 Feb;64(1):37-41. doi: 10.1016/s0166-0934(96)02142-8.
An in vitro quantal assay (TCID50) for a non-occluded baculo-like virus isolate from naturally infected Penaeus japonicus obtained from China and experimentally infected P. stylirostris was developed using primary shrimp lymphoid cell cultures in Primaria 24-well tissue culture plates. The virus caused cytopathogenic effect (CPE) in the cell cultures as early as 2 day post-infection (p.i.). Initially, the cells rounded up and finally detached from the culture vessel as the infection progressed. At the present time, there is no established quantitative in vitro cell culture protocol for the assay of this baculo-like virus which has been reported by our laboratory to be highly pathogenic for P. stylirostris and P. vannamei, the two species of penaeid shrimp commercially cultured in Hawaii and the Western hemisphere. This quantal assay thus provides a simple and convenient method for the detection and assay of infectious virus in cultured penaeid shrimp.
利用Primaria 24孔组织培养板中的对虾原代淋巴细胞培养物，开发了一种针对从中国自然感染的日本对虾以及实验感染的斯氏对虾中分离出的一种非包涵体杆状样病毒的体外定量测定法（半数组织培养感染剂量，TCID50）。该病毒在感染后第2天（p.i.）就最早在细胞培养物中引起细胞病变效应（CPE）。最初，细胞变圆，随着感染进展最终从培养容器中脱落。目前，对于这种杆状样病毒的测定，尚无已确立的体外定量细胞培养方案，据我们实验室报告，该病毒对在夏威夷和西半球商业化养殖的两种对虾——斯氏对虾和凡纳滨对虾具有高致病性。因此，这种定量测定法为养殖对虾中传染性病毒的检测和测定提供了一种简单便捷的方法。
