Equipe de Virology, FEPAGRO Animal Health--Instituto de Pesquisas Veterinárias Desidério Finamor, Eldorado do Sul, Rio Grande do Sul, Brazil.
PLoS One. 2011 Mar 2;6(3):e17501. doi: 10.1371/journal.pone.0017501.
Torque teno sus virus (TTSuV), a member of the family Anelloviridae, is a single-stranded, circular DNA virus, widely distributed in swine populations. Presently, two TTSuV genogroups are recognized: Torque teno sus virus 1 (TTSuV1) and Torque teno sus virus 2 (TTSuV2). TTSuV genomes have been found in commercial vaccines for swine, enzyme preparations and other drugs containing components of porcine origin. However, no studies have been made looking for TTSuV in cell cultures. In the present study, a search for TTSuV genomes was carried out in cell culture lineages, in sera used as supplement for cell culture media as well as in trypsin used for cell disaggregation. DNA obtained from twenty-five cell lineages (ten from cultures in routine multiplication and fifteen from frozen ampoules), nine samples of sera used in cell culture media and five batches of trypsin were examined for the presence of TTSuV DNA. Fifteen cell lineages, originated from thirteen different species contained amplifiable TTSuV genomes, including an ampoule with a cell lineage frozen in 1985. Three cell lineages of swine origin were co-infected with both TTSuV1 and TTSuV2. One batch of trypsin contained two distinct TTSuV1 plus one TTSuV2 genome, suggesting that this might have been the source of contamination, as supported by phylogenetic analyses of sequenced amplicons. Samples of fetal bovine and calf sera used in cell culture media did not contain amplifiable TTSuV DNA. This is the first report on the presence of TTSuV as contaminants in cell lineages. In addition, detection of the viral genome in an ampoule frozen in 1985 provides evidence that TTSuV contamination is not a recent event. These findings highlight the risks of TTSuV contamination in cell cultures, what may be source for contamination of biological products or compromise results of studies involving in vitro multiplied cells.
扭结藤病毒(TTSuV)是圆环病毒科的一个成员,是一种单链、环状 DNA 病毒,广泛分布于猪群中。目前,已识别出两种 TTSuV 基因群:扭结藤病毒 1(TTSuV1)和扭结藤病毒 2(TTSuV2)。TTSuV 基因组已在猪用商业疫苗、酶制剂和其他含有猪源成分的药物中被发现。然而,目前还没有研究在细胞培养物中寻找 TTSuV。在本研究中,我们在细胞培养系、细胞培养培养基中使用的血清以及用于细胞分散的胰蛋白酶中搜索 TTSuV 基因组。我们从 25 个细胞系(10 个来自常规繁殖培养,15 个来自冷冻安瓿)、9 个细胞培养培养基中使用的血清样本和 5 批胰蛋白酶中提取 DNA,以检测 TTSuV DNA 的存在。15 个细胞系,源自 13 个不同的物种,含有可扩增的 TTSuV 基因组,包括一个于 1985 年冷冻的安瓿中的细胞系。3 个猪源细胞系同时感染了 TTSuV1 和 TTSuV2。一批胰蛋白酶含有两个不同的 TTSuV1 加一个 TTSuV2 基因组,这表明这可能是污染的来源,序列分析支持了这一假设。细胞培养中使用的胎牛和小牛血清样本不含有可扩增的 TTSuV DNA。这是首次报道 TTSuV 作为细胞系的污染物。此外,在 1985 年冷冻的安瓿中检测到病毒基因组,这表明 TTSuV 污染不是最近发生的事件。这些发现强调了 TTSuV 污染在细胞培养中的风险,这可能是生物制品污染的来源,或者影响涉及体外繁殖细胞的研究结果。