β-actin实时荧光PCR法的建立和在临床试验中的应用  

Establishment of β-actin Real-time Fluorescence PCR and Its Application in Clinical Trials

作　　者：权娅茹[1] 陈震[1] 吉尚志 邱平[1] 李长贵[1] Quan Yaru;Chen Zhen;Ji Shangzhi;Qiu Ping;Li Changgui(National Institutes for Food and Drug Control,Beijing 102629,China;Beijing Wantai Biological Pharmaceutical Co.Ltd.,Beijing 102206,China)

机构地区：[1]中国食品药品检定研究院,北京102629 [2]北京万泰生物药业股份有限公司,北京102206

出　　处：《中国药事》2022年第5期535-540,共6页Chinese Pharmaceutical Affairs

摘　　要：目的:建立β-actin实时荧光聚合酶链反应(PCR)法,用于水痘减毒活疫苗和带状疱疹减毒活疫苗临床试验中样本采集有效性的判定,为临床试验中疫苗安全性判定和保护率的准确计算提供保障。方法:以β-actin质粒、水痘-带状疱疹病毒(VZV)为模板进行β-actin实时荧光PCR法体系的优化和灵敏度、特异性和重复性的验证及Cutoff值的确定。结果:建立的β-actin实时荧光PCR法特异性和重复性好,灵敏度为10 cps·μL^(-1)。临床样本β-actin扩增成功率99.9%(1746份样本中只有1份β-actin扩增阴性)。结论:建立的β-actin实时荧光PCR法可满足临床试验中样本采集有效性的判定。Objective: To establish a β-actin real-time fluorescence PCR in order to determine the validity of sample collection in clinical trials of live attenuated varicella vaccine and live attenuated herpes zoster vaccine and to guarantee vaccine safety and accurate calculation of vaccine protection rate in clinical trials. Methods:The β-actin real-time fluorescence PCR system was optimized, and the sensitivity, specificity and repeatability were verified, and the Cutoff value was determined by using the β-actin plasmid and varicella zoster virus(VZV)as templates. Results: The established β-actin real-time fluorescence PCR method has demonstrated good specificity and reproducibility, and the sensitivity was 10 cps·μL^(-1). The success rate of β-actin amplification in clinical samples reached 99.9%(only 1 β-actin amplification was negative among 1746 samples). Conclusion:The established β-actin real-time fluorescence PCR method can satisfy the confirmation of the validity of sample collection in clinical trials.

关 键 词：聚合酶链反应 β-actin质粒 灵敏度 特异性 样本采集有效性 水痘减毒活疫苗 带状疱疹减毒活疫苗 

分 类 号：R917[医药卫生—药物分析学]