摘要
目的建立一种人星状病毒TaqMan实时荧光定量RT-PCR(real-time reverse transcription-polymerase chain reaction)快速检测方法。方法根据人星状病毒ORF1b基因的保守序列设计扩增引物和特异性荧光探针,建立星状病毒TaqMan实时荧光定量RT-PCR快速检测方法,并对该方法的特异性、灵敏度和稳定性进行评价,同时应用该方法对实际临床样本中的星状病毒进行检测。结果所建立的人星状病毒TaqMan实时荧光定量RT-PCR检测方法具有良好的特异性和稳定性,灵敏度可达10^(2)拷贝/μl,对星状病毒临床样本检测后,检出率达100%。结论本文所建立的人星状病毒TaqMan实时荧光定量RT-PCR法具有简单快速、灵敏度高、特异性强、稳定性好的特点,可应用于临床星状病毒的病原检测和流行病学调查。
Objective To establish a rapid detection method for human astrovirus based on TaqMan-probe real-time fluorescent quantitative reverse transcription-polymerase chain reaction(RT-PCR).Methods According to the conservative sequence of human astrovirus ORF1b gene,we designed the amplification primers and specific fluorescent probe to establish the human astrovirus TaqMan real-time fluorescent quantitative RT-PCR rapid detection method.The specificity,sensitivity and stability of the method were evaluated.We also used this method to detect human astrovirus in clinical samples.Results The established human astrovirus TaqMan real-time fluorescent quantitative RT-PCR detection method has good specificity and repeatability for human astrovirus,and the sensitivity can reach 102 copies/μl.After testing the clinical samples,the detection rate of human astrovirus by our method was 100%.Conclusions The human astrovirus TaqMan real-time fluorescent quantitative RT-PCR detection method established in this study is simple,rapid,sensitive,specific and stable.It can be used for clinical human astrovirus detection and epidemiological investigation.
作者
杜悦
田赛
李银霞
刘鸿博
邱少富
段广才
Du Yue;Tian Sai;Li Yinxia;Liu Hongbo;Qiu Shaofu;Duan Guangcai(College of Public Health,Zhengzhou University,Zhengzhou 450001,China;Chinese PLA Center for Disease Control and Prevention,Beijing 100000,China)
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
2021年第3期195-200,共6页
Chinese Journal of Microbiology and Immunology
基金
"十三五"国家重点研发计划项目子课题(2017YFC1600105)
国家科技重大专项(2018ZX10301-407)。
