摘要
目的验证目前国内关于猴痘病毒提取试剂、扩增试剂以及实时荧光定量聚合酶链式反应(real-time quantitativepolymerase chain reaction,RT-qPCR)仪的检出效率、灵敏度差异以及提取扩增组合的比对分析。方法根据设计的正交试验方案,采用RT-qPCR对提取的核酸进行扩增。结果本次试验中不同的DNA提取试剂的提取效率的差异无统计学意义(F=12.855,P=0.070),扩增试剂扩增循环数的差异(F=49.832,P=0.019)和RT-qPCR仪对荧光捕捉信号的差异(F=36.355,P=0.027)均存在统计学意义。结论目前国内猴痘病毒的提取和扩增试剂盒可以有效地检出该病毒,试剂和仪器的组合对检出效率存在差异。通过验证试验,发现更具有检测优势的组合,为之后国内应对猴痘病毒检测及疫情防控工作提供参考。
Objective To investigate the differences in detection efficiency and sensitivity of current domestic monkeypox virus extraction reagents,amplification reagents and real-time quantitative polymerase chain reaction(RT-qPCR)instruments,and to compare among different extraction and amplification combinations.Methods The extracted nucleic acid of monkeypox virus were amplified using RT-qPCR,according to the designed orthogonal test protocol.Results There was no significance in the extraction efficiency of different DNA extraction reagents(F=12.855,P=0.07),while the differences were statistically differences in the number of amplification cycles of amplification reagents(F=49.832,P=0.019)and in the fluorescence signal capture of RT-qPCR instruments(F=36.355,P=0.027).Conclusion The current domestic extraction and amplification kits for monkeypox virus can effectively detect the virus,but various reagent and instrument combinations show different detection efficiency.Through the validation test,a more advantageous combination is found,which provides a reference for domestic response to monkeypox virus detection and epidemic prevention and control.
作者
宗华
孙巧
李彩云
殷静
罗青梅
王梦佳
ZONG Hua;SUN Qiao;LI Caiyun;YIN Jing;LUO Qingmei;WANG Mengjia(Department of Microbiological Laboratory,Nan’an District Center for Disease Control and Prevention,Chongqing,400067;School of Public Health,Chongqing Medical University,Chongqing,400016,China)
出处
《陆军军医大学学报》
CAS
CSCD
北大核心
2022年第23期2440-2445,共6页
Journal of Army Medical University
作者简介
通信作者:孙巧,E-mail:378940993@qq.com
