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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RT_RAA快速检测方法的建立及应用-宁波国旅&宁波检验检疫

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Abstract: A recombinase-aided amplification (RAA)-based nucleic acid detection method for Middle East Respiratory Syndrome Coronavirus (MERS-CoV) has been established. In this study, we designed and synthesized specific RAA primers and probes targeting the MERS-CoV gene, and prepared a positive control consisting of MERS-CoV pseudovirus particles. Through systematic optimization of experimental conditions, we developed a rapid, sensitive, and highly specific RT-RAA assay for MERS-CoV detection. The new method was then compared with a commercially available fluorescent quantitative RT-PCR assay. Furthermore, clinical validation was performed using pharyngeal swab samples from China’s first imported MERS-CoV case originating from South Korea, along with other respiratory virus samples and inactivated MERSCoV proficiency testing materials provided by UK QC-MD. The results demonstrated that the established RT-RAA method exhibits a detection sensitivity of 10 copies, surpassing the sensitivity of our lab’s previously developed fluorescent RT-PCR assay (100 copies). Additionally, the assay significantly reduces the detection time to just 48–13.6 minutes, compared to approximately 90 minutes required by the fluorescent RT-PCR method. Importantly, the RT-RAA assay yielded positive results when applied to the MERS-CoV pseudovirus particles, while all eight other control respiratory viruses tested negative. Clinical samples analyzed using this method also showed concordance with their actual diagnostic outcomes. Overall, the RT-RAA assay for MERS-CoV established in this study is highly sensitive, specific, and rapid, making it an ideal tool for on-site, point-of-care diagnosis and epidemiological surveillance of MERS-CoV infections.

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新型冠状病毒核酸荧光型RT-RAA检测方法的建立及其评价

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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Previous page

新型冠状病毒核酸荧光型RT-RAA检测方法的建立及其评价

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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