Viral RNA Extraction and Real-Time RT-PCR Detection

Viral RNA was extracted from cultured viruses using a QIAamp Viral RNA Extraction Mini Kit (Qiagen, Valencia, CA) according to the manufacturer instructions. Viral RNA was detected by real-time reverse transcription-polymerase chain reaction (real-time RT-PCR) as previously described3 (link)13 (link). Briefly, a 185-bp amplicon was generated and detect by a TaqMan probe labeled with FAM and Blackhole Quencher. The amplification was carried out using an ABI 7500 real-time system (Life Technologies, NY) and the results analyzed with the instrument SDS software (version 2.0.1).

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Zhu Z., Rivailler P., Abernathy E., Cui A., Zhang Y., Mao N., Xu S., Zhou S., Lei Y., Wang Y., Zheng H., He J., Chen Y., Li C., Bo F., Zhao C., Chen M., Lu P., Li F., Gu S., Gao H., Guo Y., Chen H., Feng D., Wang S., Tang X., Lei Y., Feng Y., Deng L., Gong T., Fan L., Xu W, & Icenogle J. (2015). Evolutionary analysis of rubella viruses in mainland China during 2010–2012: endemic circulation of genotype 1E and introductions of genotype 2B. Scientific Reports, 5, 7999.

Publication 2015

QIAamp Viral RNA Extraction Mini Kit ABI 7500 real-time system

Real time pcr Reverse transcription Viral rna Viruses

Corresponding Organization :

Other organizations : National Institute for Viral Disease Control and Prevention, Centers for Disease Control and Prevention, Tianjin Centers for Disease Control and Prevention, Sichuan Center for Disease Control and Prevention, Gansu Provincial Center for Disease Control and Prevention, Beijing Center for Disease Prevention and Control, Ningxia Center for Diseases Prevention and Control, Henan Provincial Center for Disease Control and Prevention, Hubei Provincial Center for Disease Control and Prevention, Zhejiang Center for Disease Control and Prevention

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Viral RNA detection by real-time reverse transcription-polymerase chain reaction (real-time RT-PCR)

control variables

Viral RNA extraction using QIAamp Viral RNA Extraction Mini Kit (Qiagen, Valencia, CA)
Amplification carried out using an ABI 7500 real-time system (Life Technologies, NY)
Results analyzed with the instrument SDS software (version 2.0.1)

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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