Innovative Technology

Innovation

Focusing on the research and development and application of isothermal nucleic acid amplification technology

中东呼吸综合征冠状病毒重组酶介导核酸检测方法的建立-宁波国旅&宁波检验检疫

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Abstract: Objective To establish a rapid, sensitive, and specific reverse-transcription recombinase-aided assay (RT-RAA) for the detection of Middle East Respiratory Syndrome Coronavirus (MERS-CoV). Methods Specific RAA primers and probes were designed and synthesized based on the conserved sequences of the MERS-CoV gene. After optimizing reaction conditions, the RT-RAA assay was performed using a 10-fold serially diluted positive control standard—pseudo-viral particles generated from a third-generation lentiviral vector—as the template. The assay’s specificity, sensitivity, and applicability to clinical samples were evaluated. Results The established RT-RAA method demonstrated a detection limit of 10 copies/mL for the MERS-CoV pseudo-viral particle standard, outperforming the real-time fluorescent RT-PCR method (10 copies/mL). Notably, this RT-RAA assay showed clear amplification specifically targeting MERS-CoV, while yielding negative results for four other respiratory viruses, including influenza A H1N1, influenza B, respiratory syncytial virus, and rhinovirus. Furthermore, the assay successfully detected both the clinical sample from China’s first imported MERS case and the positive inactivated sample provided by the UK OCMD proficiency testing program, confirming its accuracy. Importantly, the assay’s total turnaround time (20 minutes) was significantly shorter than that of real-time fluorescent RT-PCR (90 minutes). Conclusion The developed MERS-CoV RT-RAA assay is highly sensitive, specific, and rapid, making it suitable for point-of-care detection of MERS-CoV infections as well as epidemiological investigations.

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中东呼吸综合征冠状病毒RT_RAA快速检测方法的建立及应用-宁波国旅&宁波检验检疫

重组酶介导恒温扩增快速检测人鼻病毒的方法学研究_陈淑丹

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Previous page

中东呼吸综合征冠状病毒RT_RAA快速检测方法的建立及应用-宁波国旅&宁波检验检疫

Next page

重组酶介导恒温扩增快速检测人鼻病毒的方法学研究_陈淑丹