评估下一代测序性能,以在体外检测生物产品中的病毒
Takamasa Hirai1, Kiyoko Kataoka1, Yuzhe Yuan2
1Division of Cell-Based Therapeutic Products, National Institute of Health Sciences, Kanagawa, Japan.
概括
下一代测序 (NGS) 提供了一种敏感的方法来检测生物制品中的病毒污染. 这项研究表明,NGS可以检测到只有一份病毒序列副本,支持其在病毒安全测试中的使用.
科学领域:
- 生物技术是生物技术.
- 病毒学 病毒学
- 分子生物学分子生物学
背景情况:
- 下一代测序 (NGS) 是一种大规模并行测序技术.
- 国际协调理事会 (ICH) Q5A (R2) 草案建议NGS作为生物制品传统体外病毒检测的潜在替代品或补充.
- 检测偶然的病毒污染对生物安全至关重要.
研究的目的:
- 评估下一代测序 (NGS) 在体外检测病毒的性能.
- 评估短读和长读NGS对病毒污染的敏感性和检测极限.
- 确定NGS作为一种用于检测生物制品中偶然性剂的方法的潜力.
主要方法:
- 5型腺病毒 (Ad5) 被用作模型病毒,并注射到Vero细胞中.
- 总RNA从感染和未感染的Vero细胞中提取.
- 样品被连续稀释并使用短读和长读NGS平台进行分析.
主要成果:
- 短读和长读NGS方法都显示了可比的检测极限.
- NGS方法具有高度灵敏度,能够检测每次测试至少有一副病毒序列.
- 在多重NGS测定中,交叉污染的可能性被确定为错误阳性的风险.
结论:
- 下一代测序 (NGS) 是一种敏感且快速的技术,用于检测生物制品中的病毒污染.
- 作为生物制药行业病毒安全测试的有价值工具,NGS显示出前景.
- 需要进一步验证以减轻多重NGS应用中的交叉污染等风险.
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