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A next-generation sequencing based method for determining genetic stability in Clostridium tetani vaccine strains.
Biologicals ( IF 1.7 ) Pub Date : 2020-02-11 , DOI: 10.1016/j.biologicals.2020.02.001
Jeroen L A Pennings 1 , Eric Abachin 2 , Sylvie Uhlrich 2 , Raphaël Esson 2 , Laurent Mallet 2 , Rob J Vandebriel 1
Affiliation  

Production of tetanus and other clostridial vaccines highly depends on the stable and reproducible production of high toxin levels. This creates a need to ensure the genetic stability of seed strains. We developed a two-stage method for improved assessment of the genetic stability of Clostridium seed strains. This method is based on next-generation sequencing (NGS) of strain DNA and mapping the sequence reads to a reference sequence. The output allows analysis of global genome consistency followed, if necessary, by detailed expert judgement of potential deviations at the gene level. The limit of detection of our method is an order of magnitude better than that of the currently established pulsed-field gel electrophoresis (PFGE). Improved genetic characterization of bacterial seed lots will have a positive impact on the characterization of the production process. This will be a first step towards applying the consistency approach to vaccine batch release of established vaccines. This can contribute to the reduction and ultimately replacement of routinely used animal tests in vaccine production. This work was carried out as part of the Innovative Medicines Initiative 2 (IMI2) project VAC2VAC (Vaccine batch to vaccine batch comparison by consistency testing).



中文翻译:

基于下一代测序的方法来确定破伤风梭菌疫苗菌株的遗传稳定性。

破伤风和其他梭菌疫苗的生产高度依赖于高毒素水平的稳定和可重复生产。这就需要确保种子菌株的遗传稳定性。我们开发了一种两阶段方法来改进梭菌的遗传稳定性评估种子株。该方法基于菌株DNA的下一代测序(NGS),并将读取的序列映射到参考序列。输出结果允许对全局基因组一致性进行分析,必要时还可以根据专家对基因水平潜在偏差的详细判断。我们方法的检测极限比当前建立的脉冲场凝胶电泳(PFGE)的检测极限好一个数量级。改善细菌种子批次的遗传特性将对生产过程的特性产生积极影响。这将是将一致性方法应用于已建立疫苗的疫苗批量释放的第一步。这可以有助于减少并最终替代疫苗生产中常规使用的动物试验。

更新日期:2020-02-11
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