高通量测序技术在生物技术产品病毒安全控制中的应用

Application of High Throughput Sequencing Technique in Viral Safety Control of Biotechnology Products

第二代测序是以边合成边测序为核心理念的高通量测序技术,检测通量高、速度快、低丰度序列检出能力强等特点决定了其在生物技术产品的病毒安全控制方面具有重要的应用价值。2019年,国际人用药物协调会议的新加坡面对面会议期间,Q5A(R2)指导原则《来源于人或动物细胞系的生物技术产品的病毒安全性评价》专家工作组在专题讨论中明确,考虑高通量测序技术在原始材料的病毒检定,特别是在针对非预期病毒的检定方面具有明显优势,因此,鼓励研究人员采用高通量测序技术对生物技术产品的内、外源因子进行检定,并将在此次修订稿中针对高通量测序技术提出原则性的技术要求。另一方面,在具有广阔应用前景的同时,高通量测序技术在病毒检测的实际应用中还存在一些挑战,比如样本的预处理、参考数据库的建立和研究结果的判定等,均会对检测的最终结果产生影响。文章从测序技术的发展历程,药典和指南,应用实例,挑战与对策,思考与讨论5个方面展开讨论,旨在探索技术要求的科学性,引起业界和监管机构的共同思考。

The second generation sequencing is a high-throughput sequencing(HTS)technique based on the core concept of sequencing while synthesizing at the same time.The characteristics of high detection throughput,fast speed,and strong ability to detect low-abundance sequences determine that it has important application value in the viral safety control of biotechnology products.During the Singapore face-to-face meeting of the International Council for Harmonization in 2019,the expert working group(EWG)on the Q5 A(R2)guideline"Viral Safety Evaluation of Biotechnology Products Derived from Cell Lines of Human or Animal Origin"made it clear in the panel discussion that considering HTS technology has a clear advantage in virus detection of initial raw materials,especially in the detection of unexpected viruses.Therefore,researchers are encouraged to use high-throughput sequencing technology to verify the internal and external factors of biotechnology products,and the EWG will put forward high level technical requirements for HTS technology in this revised version of Q5 A guideline.On the other hand,while it has a broad application prospect,there are still some challenges in the practical application of HTS technology in virus detection,such as the preprocessing of samples,the establishment of reference database and the determination of research results,etc.,which will have a significant impact on the final results of detection.In order to explore the scientific nature of the technical requirements and arouse the common thinking of the industry and regulatory authorities,this article discusses from five aspects,which are the development of sequencing technique,pharmacopoeia and guidelines,application examples,challenges and countermeasures,thinking and discussion.