利用逆转录环介导恒温扩增技术建立新型冠状病毒核酸快速检测方法

The Rapid Detection of Nucleic Acid in SARS-CoV-2 by Reverse Transcription Loop Mediated Isothermal Amplification

目的利用逆转录环介导恒温扩增技术建立新型冠状病毒核酸快速检测方法。方法针对新型冠状病毒的特征核壳蛋白N基因和开放阅读框1ab(ORF1ab)基因序列保守区设计引物,获得了RT-LAMP检测体系,并评价该体系的特异性、最低检测限、抗干扰能力和最佳反应时间。结果经检测新型冠状病毒核酸标准物质,其结果与预期相符,证实了该检测体系的可行性。结论利用逆转录环介导恒温扩增技术快速检测新型冠状病毒核酸的反应体系特异性好、灵敏度高,适用于边境条件有限地区。

Objective To establish a rapid detection method for the nucleic acid of severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)based on the reverse transcription loop mediated isothermal amplification(RT-LAMP).Method The RT-LAMP primers were designed according to the conservative region of N and ORF1ab gene of SARS-CoV-2.And the specificity,the minimum detection limit,the anti-interference ability and the best reaction time of the system were evaluated.Result Artificial synthesized SARS-CoV-2 nucleic acid samples were tested,and the results were consistent with the expected results.It confirmed the feasibility of the detection system.Conclusion The reaction system using RT-LAMP for the rapid detection of SARS-CoV-2 nucleic acid has the high specificity and sensitivity,and is suitable for the border areas with limited conditions.