基于毛细管电泳的快速检测七种人冠状病毒技术方法的建立

Rapid Simultaneous Detection of Seven Human Coronaviruses Using Capillary Electrophoresis

近来,一种新型冠状病毒(SARS-CoV-2)引发的COVID-19突发疫情,给全球公众健康和社会经济构成严重威胁。SARS-CoV-2成为继人冠状病毒229E(Human coronavirus 229E,HCoV-229E)、人冠状病毒OC43(Human coronavirus OC43,HCoV-OC43)、严重急性呼吸综合征冠状病毒(Severe acute respiratory syndrome coronavirus,SARS-CoV)、人冠状病毒NL63(Human coronavirus NL63,HCoV-NL63)、人冠状病毒HKU1(Human coronavirus HKU1,HCoV-HKU1)和中东呼吸综合征冠状病毒(Middle East respiratory syndrome coronavirus,MERS-CoV)后第七种感染人类的冠状病毒。本研究以高分辨毛细管电泳技术为基础,针对七种人冠状病毒基因保守区分别设计特异性引物对,经常规PCR扩增后,通过具备单碱基差异分辨率的毛细管电泳分析,实现快速检测七种人冠状病毒的目标。通过构建基于毛细管电泳的人冠状病毒分子靶标,实现同时快速精准鉴定七种人冠状病毒的目的。本研究建立的人冠状病毒毛细管电泳快速检测技术方法具有极高灵敏性和精确性,分辨率高而且特异性好,操作简便成本低廉,为人冠状病毒的临床诊断、口岸快速检测等提供了新的技术支持。

SARS-CoV-2 infection causes coronavirus disease 2019(COVID-19).The latter is wreaking havoc on health care and economic systems worldwide.SARS-CoV-2 is the seventh coronavirus known to infect humans.Rapid diagnoses are vital for the prevention and therapeutic intervention of coronavirus infections in humans.First,we designed specific primer pairs targeting the conserved regions of seven human coronavirus genes,respectively.After amplification by conventional reverse transcription-polymerase chain reaction(RT-PCR),PCR products were analyzed by capillary electrophoresis(CE)with single-base differential resolution for rapid detection of seven human coronaviruses,respectively.Through identification of molecular markers of coronaviruses based on CE,we established a novel assay for rapid and accurate identification of seven human coronaviruses simultaneously.The assay using CE had high sensitivity,accuracy,resolution and specificity;the operation was simple and inexpensive.Thus,this novel assay provides new technical support for detection of human coronaviruses in the clinic and ports of entry into China.