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Tytuł:
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[Development and verification of reference nucleic acid materials of H9N2 influenza viruses by real-time RT-PCR].
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Autorzy:
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Song J; Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China.; University of Chinese Academy of Sciences, Beijing 100049, China.
Li C; China Institute of Veterinary Drug Control, Beijing 100081, China.
Li J; Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China.; University of Chinese Academy of Sciences, Beijing 100049, China.
Zhang S; Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China.
Fan W; Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China.
Liu L; Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China.
Jia H; Beijing Haidian Foreign Language Experiment School, Beijing 100195, China.
Yu A; Beijing Haidian Foreign Language Experiment School, Beijing 100195, China.
Hao K; Beijing Haidian Foreign Language Experiment School, Beijing 100195, China.
Niu C; National Institute of Metrology, Beijing 100013, China.
Wang J; National Institute of Metrology, Beijing 100013, China.
Zhao Q; China Institute of Veterinary Drug Control, Beijing 100081, China.
Liu W; Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China.; University of Chinese Academy of Sciences, Beijing 100049, China.
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Źródło:
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Sheng wu gong cheng xue bao = Chinese journal of biotechnology [Sheng Wu Gong Cheng Xue Bao] 2018 Oct 25; Vol. 34 (10), pp. 1579-1586.
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Typ publikacji:
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Journal Article
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Język:
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Chinese
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Imprint Name(s):
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Original Publication: Beijing : Ke xue chu ban she,
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MeSH Terms:
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Real-Time Polymerase Chain Reaction*
Reverse Transcriptase Polymerase Chain Reaction*
Hemagglutinin Glycoproteins, Influenza Virus/*genetics
Influenza A Virus, H9N2 Subtype/*genetics
RNA, Viral/*genetics
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Contributed Indexing:
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Keywords: H9N2 subtype influenza virus; HA gene; real-time RT-PCR; reference materials
Local Abstract: [Publisher, Chinese] 从H9N2 亚型流感病毒A/chicken/Hunan/04.14 (H9N2) 核酸中扩增了HA 基因的编码序列，克隆测序后，采用体外转录方法制备RNA。用RNA 保存液稀释至含量约10⁹ copies/μL。分装后进行均匀性和稳定性检验，通过4 家实验室协作标定，取平均值作为定值结果。此外，文中建立的实时荧光定量PCR (qPCR) 快速检测技术，对临床样品进行准确检测验证，检测限可达10 个拷贝。结果表明，文中制备的核酸参考品可作为H9N2 亚型流感病毒核酸快速检测方法的阳性定量参考品。.
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Substance Nomenclature:
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0 (Hemagglutinin Glycoproteins, Influenza Virus)
0 (RNA, Viral)
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Entry Date(s):
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Date Created: 20181106 Date Completed: 20190109 Latest Revision: 20190109
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Update Code:
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20240104
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DOI:
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10.13345/j.cjb.180034
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PMID:
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30394025
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The HA gene of H9N2 influenza virus (A/chicken/Hunan/04.14 (H9N2)) was amplified and sequenced. The RNA was synthesized by in vitro transcription. The RNA transcription solutions were diluted to 10⁹ copies/μL using the RNA storage solution. The aliquoted RNA solutions were used to evaluate the homogeneity and stability. The results were determined by the average value obtained from four independent laboratories. Furthermore, the fluorescence quantitative RT-PCR method was also developed to verify the detection accuracy of clinical samples. The detection limit of this method is approximately 10 copies. Taken together, the RNA transcription solution established in our study can used as positive standard reference for rapid detection of H9N2 influenza virus.
