Torque Teno Sus Virus (TTSuV) in Cell Cultures and Trypsin

PLoS One

Ana Cláudia Franco¹  Thais Fumaco Teixeira²  Samuel Paulo Cibulski²  Carine Lidiane Holz²  Ana Paula Muterle Varela²  Paulo Michel Roehe³  Diogenes Dezen³ 

[1] CIRAD, Départament Systèmes Biologiques, UR-15, Campus International de Baillarguet, Montpellier, France;Equipe de Virology, FEPAGRO Animal Health - Instituto de Pesquisas Veterinárias Desidério Finamor (IPVDF), Eldorado do Sul, Rio Grande do Sul, Brazil;Virology Laboratory, Department of Microbiology, Immunology and Parasitology, Instituto de Ciências Básicas da Saúde, Universidade Federal do Rio Grande do Sul, Porto Alegre, Rio Grande do Sul, Brazil

关键词: Serine proteases;    Cell cultures;    Anelloviruses;    Polymerase chain reaction;    Swine;    Mammalian genomics;    DNA extraction;    Kidneys;   

DOI  :  10.1371/journal.pone.0017501

学科分类：医学（综合）

来源: Public Library of Science

【 摘 要 】

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.

【 授权许可】

CC BY