Development and verification of reference nucleic acid materials of H9N2 influenza viruses by real-time RT-PCR.

Jie Song; Cui LI; Jing LI; Shuang Zhang; Wenhui Fan; Lirong Liu; Hongyi Jia; Aibi YU; Ke Hao; Chunyan Niu; Jing Wang; Qizu Zhao; Wenjun Liu

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Development and verification of reference nucleic acid materials of H9N2 influenza viruses by real-time RT-PCR.

Author: Jie SONG ¹ ; Cui LI ² ; Jing LI ¹ ; Shuang ZHANG ¹ ; Wenhui FAN ¹ ; Lirong LIU ¹ ; Hongyi JIA ³ ; Aibi YU ³ ; Ke HAO ³ ; Chunyan NIU ⁴ ; Jing WANG ⁴ ; Qizu ZHAO ² ; Wenjun LIU ¹
Author Information

1. Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China.
2. China Institute of Veterinary Drug Control, Beijing 100081, China.
3. Beijing Haidian Foreign Language Experiment School, Beijing 100195, China.
4. National Institute of Metrology, Beijing 100013, China.
Publication Type:Journal Article
Keywords: H9N2 subtype influenza virus; HA gene; real-time RT-PCR; reference materials
From: Chinese Journal of Biotechnology 2018;34(10):1579-1586
CountryChina
Language:Chinese
Abstract: 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.