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利用劳斯肉瘤病毒(RSV)阴性细胞系检测内源性和外源性淋巴白血病病毒及鸡辅助因子

Assays for endogenous and exogenous lymphoid leukosis viruses and chick helper factor with RSV(--) cell lines.

作者信息

Crittenden L B, Eagen D A, Gulvas F A

出版信息

Infect Immun. 1979 May;24(2):379-86. doi: 10.1128/iai.24.2.379-386.1979.

Abstract

Japanese quail cells transformed by the envelope-defective Bryan high-titer strain of Rous sarcoma virus [R(-)Q] were used as a source of the Rous sarcoma virus genome in three kinds of assays. (i) The simplest and most sensitive assay for infectious, endogenous viruses of the chicken belonging to subgroup E involved infection of a mixture of R(-)Q cells and turkey cells with the sample and assay of supernatants of these cells for focus formation on subgroup E susceptible cells. (ii) Inactivated Sendai virus-induced fusion of R(-)Q cells with live test cells was found to be a specific method for detection of chick helper factor. Focus formation by supernatant of the fused cells on subgroup E susceptible cells was correlated with the presence of subgroup E envelope glycoprotein on the plasma membranes of test cells. Whole blood cells as well as fibroblasts could be used in this assay. (iii) A method of assay for exogenous lymphoid leukosis viruses in which mixed cultures of R(-)Q cells and C/E cells and assay of supernatants for focus formation on C/E cells was as sensitive as assays presently used for exogenous lymphoid leukosis virus. Because no infectious Rous sarcoma virus was used as part of the procedure, the assays for infectious virus described here yielded pure pseudotypes of the input virus, an advantage for determining purity and subgroup of the input virus.

摘要

在三种检测中,包膜缺陷型劳氏肉瘤病毒布莱恩高滴度株[R(-)Q]转化的日本鹌鹑细胞被用作劳氏肉瘤病毒基因组的来源。(i) 对于属于E亚群的鸡的传染性内源性病毒,最简单且最灵敏的检测方法是用样品感染R(-)Q细胞和火鸡细胞的混合物,并检测这些细胞的上清液在E亚群敏感细胞上形成的病灶。(ii) 发现仙台病毒灭活诱导R(-)Q细胞与活的测试细胞融合是检测鸡辅助因子的一种特异性方法。融合细胞上清液在E亚群敏感细胞上形成病灶与测试细胞质膜上E亚群包膜糖蛋白的存在相关。全血细胞和成纤维细胞均可用于该检测。(iii) 一种检测外源性淋巴细胞白血病病毒的方法,其中R(-)Q细胞和C/E细胞的混合培养以及检测上清液在C/E细胞上形成的病灶,其灵敏度与目前用于检测外源性淋巴细胞白血病病毒的检测方法相同。由于该检测过程未使用传染性劳氏肉瘤病毒,这里描述的传染性病毒检测产生了输入病毒的纯假型,这对于确定输入病毒的纯度和亚群是一个优势。

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