人民卫生出版社系列期刊
ISSN 2096-2738 CN 11-9370/R

中国科技核心期刊(中国科技论文统计源期刊)
2020《中国学术期刊影响因子年报》统计源期刊

新发传染病电子杂志 ›› 2020, Vol. 5 ›› Issue (2): 95-99.

• 论著 • 上一篇    下一篇

基于微流控芯片技术的5种呼吸道病毒核酸快速荧光qPCR检测方法的建立

任琴1#, 张星艳1#, 杜寿文1, 王宇航1, 杨桂林1, 林琳华2, 戴少波1, 李体远1, 金宁一3, 李昌3, 吴诗品1   

  1. 1.暨南大学第二临床医学院,深圳市人民医院感染内科,广东 深圳 518020;
    2.暨南大学第二临床医学院,深圳市人民医院产科,广东 深圳 518020;
    3.军事科学院军事医学研究院军事兽医研究所,省部共建吉林省人兽共患病预防与控制重点实验室,长春 130062
  • 收稿日期:2020-02-15 出版日期:2020-02-20 发布日期:2020-06-19
  • 通讯作者: 李昌,Email:lichang78@163.com;吴诗品,Email: wupoem@126.com.
    #:共同第一作者
  • 基金资助:
    深圳市政府卫生“三名工程”专项基金(SZSM201612034)

Establishment of rapid qPCR method for detection of five respiratory viruses based on microfluidic chip technology

Ren Qin1#, Zhang Xingyan1#, Du Shouwen1, Wang Yuhang1, Yang Guilin1, Lin Linhua2, Dai Shaobo1, Li Tiyuan1, Jin Ningyi3, Li Chang3, Wu Shipin1   

  1. 1. Department of Infectious Diseases, the Second Clinical Medical College of Jinan University, Shenzhen People's Hospital, Guangdong Shenzhen 518020, China;
    2. Department of Obstetrics, the Second Clinical Medical College, Shenzhen People's Hospital of Jinan University, Guangdong Shenzhen 518020, China;
    3. Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary Medicine, Academy of Military Medical Sciences, Academy of Military Sciences, Changchun 130062, China
  • Received:2020-02-15 Online:2020-02-20 Published:2020-06-19

摘要: 目的 建立一种快速鉴别诊断5种重要呼吸道病毒(甲型流感病毒、乙型流感病毒、H1N1和H3N2亚型流感病毒及麻疹病毒)的检测方法。方法 基于微流控芯片技术,设计合成针对5种呼吸道病毒的特异性探针引物,利用快速反应体系及创新性温控模块研制的极速荧光定量PCR仪,评估探针引物的特异性、灵敏性及检测体系的重复性,并用临床样本进行测试。结果 设计合成的5种病毒的探针引物具有较高的特异性和灵敏性,建立的检测体系可在5min内完成病毒核酸的制备,30min内获得检测结果,且具备良好的重复性。结论 建立了5种呼吸道病毒核酸快速定量检测方法,有望用于不明呼吸道病毒感染的鉴别诊断及现场快速检测。

关键词: 呼吸道病毒, 快速定量检测, 微流控芯片技术

Abstract: Objective To establish a rapid differential diagnostic method for five respiratory viruses (influenza A virus, influenza B virus, H1N1/H3N2 subtype influenza virus and measles virus). Methods We designed and synthesized specific probe primers for the five respiratory viruses respectively. Based on the latest microfluidic chip technology, the specificity, sensitivity and repeatability of the detection system of probe primers was evaluated by a rapid fluorescent quantitative PCR instrument with the rapid reaction system and innovative temperature control module, and the method was used to test some clinical samples.Results The probe primers of five viruses designed and synthesized have high specificity and sensitivity. Virus nucleic acid was extracted in 5 minutes, and the detection results were obtained in 30 minutes and had good repeatability. Conclusion A rapid and quantitative detection method for five respiratory viruses was established, which was expected to be used in the differential diagnosis and rapid detection of unknown respiratory virus infection.

Key words: Respiratory virus, Rapid quantitative detection, Microfluidic chip technology