牛病毒性腹泻病毒（BVDV）对细胞培养物的污染。

Contamination of cell cultures with bovine viral diarrhea virus (BVDV).

作者信息

Uryvaev L V, Dedova A V, Dedova L V, Ionova K S, Parasjuk N A, Selivanova T K, Bunkova N I, Gushina E A, Grebennikova T V, Podchernjaeva R J

机构信息

D. I. Ivanovsky Institute of Virology, Ministry of Health and Social Development of Russian Federation, Moscow, Russia.

出版信息

Bull Exp Biol Med. 2012 May;153(1):77-81. doi: 10.1007/s10517-012-1648-1.

DOI:10.1007/s10517-012-1648-1

PMID:22808499

Abstract

The incidence of contamination of cell strains used in biological and virological studies and of fetal calf sera (FCS) manufactured by Russian and foreign companies used for cell culturing with noncytocidal bovine viral diarrhea virus (BVDV; Pestivirus, Flaviviridae) was analyzed. The virus was detected by reverse transcription PCR and indirect immunofluorescence with monoclonal antibodies to BVDV virion envelope glycoprotein in 25% of 117 cell strains and 45% of 35 tested FCS lots. The virus multiplied and persisted in a wide spectrum of human cell strains and in monkey, swine, sheep, rabbit, dog, cat, and other animal cells. The levels of BVDV genome RNA in contaminated cell cultures reached 10(2)-10(3) g-eq/cell and in serum samples 10(3)-10(7) g-eq/ml. These facts necessitate testing of cells and FCS for BVDV reproduced in cells without signs of infection detectable by light microscopy. The molecular mechanisms of long-term virus persistence in cells without manifestation of cell destruction are unknown.

摘要

分析了俄罗斯和外国公司生产的用于细胞培养的生物和病毒学研究中使用的细胞株以及胎牛血清（FCS）被非细胞病变性牛病毒性腹泻病毒（BVDV；瘟病毒属，黄病毒科）污染的发生率。通过逆转录聚合酶链反应（RT-PCR）以及使用针对BVDV病毒粒子包膜糖蛋白的单克隆抗体进行间接免疫荧光检测，在117个细胞株中的25%以及35个测试的FCS批次中的45%检测到了该病毒。该病毒能在广泛的人类细胞株以及猴、猪、羊、兔、狗、猫和其他动物细胞中增殖并持续存在。受污染细胞培养物中BVDV基因组RNA水平达到10² - 10³基因组当量/细胞，血清样本中为10³ - 10⁷基因组当量/毫升。这些事实使得有必要对细胞和FCS进行检测，以确定是否存在在细胞中复制但通过光学显微镜检测不到感染迹象的BVDV。病毒在细胞中无细胞破坏表现的情况下长期持续存在的分子机制尚不清楚。