Dos Santos Sergio, Lespinasse Emilie, Bonnet Baptiste, Basmaciogullari Stéphane
Sanofi R&D, 94400 Vitry-sur-Seine, France.
Mol Ther Methods Clin Dev. 2025 Aug 20;33(3):101572. doi: 10.1016/j.omtm.2025.101572. eCollection 2025 Sep 11.
Cell substrate utilized in the production of biologics intended for human use need to be cleared from mycoplasma contaminant as described in pharmacopoeia. While the gold-standard method for mycoplasma detection involves lengthy microbiology and cell-based assays, nucleic acid technologies offer the possibility to accelerate testing. In this study, we develop a qPCR assay capable of specifically detecting 11 mycoplasma species relevant to pharmacopoeias with only two primers and two hydrolysis probes, which greatly facilitates operations. While the primers cross-react with bacterial species, the specificity conferred by the hydrolysis probes allows for confident interpretation of the qPCR result and unambiguous statement regarding the presence of mycoplasma in the test article. Amplicon sequencing can further confirm the identity of contaminants. This comprehensive assay can therefore be of great help to quality control laboratories embedded in biologics production sites, which need to provide results in a timely manner and contribute to root cause analysis in case of contamination.
用于生产供人类使用的生物制品的细胞基质需要按照药典所述清除支原体污染物。虽然支原体检测的金标准方法涉及冗长的微生物学和基于细胞的检测，但核酸技术提供了加速检测的可能性。在本研究中，我们开发了一种qPCR检测方法，仅用两个引物和两个水解探针就能特异性检测与药典相关的11种支原体，这极大地简化了操作。虽然引物与细菌物种有交叉反应，但水解探针赋予的特异性使得能够可靠地解释qPCR结果，并就测试样品中支原体的存在作出明确说明。扩增子测序可以进一步确认污染物的身份。因此，这种全面的检测方法对生物制品生产现场的质量控制实验室有很大帮助，这些实验室需要及时提供结果，并在发生污染时有助于进行根本原因分析。
