Mavin S, Joss A W L, Ball J, Ho-Yen D O
Scottish Toxoplasma Reference Laboratory, Raigmore Hospital, Inverness IV2 3UJ, UK.
J Clin Pathol. 2004 Jun;57(6):609-11. doi: 10.1136/jcp.2003.013763.
To examine three lineages of Toxoplasma gondii RH strain in terms of performance in the dye test, culture, and gene expression.
Historical data (culture growth and performance in the dye test) from three lineages of RH strain tachyzoites (B, J, and Q) that had been continuously cultured in HeLa cells was assessed. Tachyzoite harvests obtained during continuous cell culture were retrieved from liquid nitrogen and cultured in HeLa cells, providing mRNA that was extracted and used to study gene expression using random amplified polymorphic DNA analysis at different stages of lineage adaptation to continuous culture.
The B and Q lineages consistently produced tachyzoites that were successfully used in the dye test and their gene expression was stable after multiple passages. The J lineage had unpredictable growth, tachyzoites unsuitable for use in the dye test, and changing gene expression with multiple passage.
This study has explained some anomalies in the performance of different stocks of T gondii, and suggests that lineages that are still evolving in cell culture should be avoided.
从染色试验、培养及基因表达方面研究弓形虫RH株的三个谱系。
评估在HeLa细胞中连续培养的RH株速殖子三个谱系（B、J和Q）的历史数据（培养生长情况及染色试验表现）。从液氮中取出连续细胞培养过程中收获的速殖子，在HeLa细胞中培养，提取mRNA，用于在谱系适应连续培养的不同阶段通过随机扩增多态性DNA分析研究基因表达。
B和Q谱系持续产生可成功用于染色试验的速殖子，且多次传代后其基因表达稳定。J谱系生长不可预测，其速殖子不适合用于染色试验，且多次传代后基因表达发生变化。
本研究解释了不同弓形虫株表现中的一些异常情况，并表明应避免使用在细胞培养中仍在进化的谱系。
