摘要
目的研究新型中药制剂对疟原虫的杀伤及对宿主的免疫影响。方法应用将多重PCR扩增和液相芯片检测技术检测埃博拉病毒(EBV)、拉沙热病毒(LFV)、裂谷热病毒(RVFV)、马尔堡病毒(MRBV)、克里米亚-刚果出血热病毒(CCHFV)、中东呼吸综合征病毒(MERSV),为相关输入性烈性传染病的检测提供一种技术储备。方法建立6种相关病毒的多重PCR扩增反应体系,分别用每种病毒特异性核酸探针与不同编码的荧光微球进行偶联,将PCR扩增产物与偶联有不同核酸探针的微球混合物进行杂交反应,建立6种病毒的液相芯片检测方法,并进行特异性和灵敏度验证。结果特异性检测结果表明不同病毒之间检测结果没有出现相互交叉干扰的现象,说明建立的6种烈性传染病病毒液相芯片检测方法用于不同病毒的检测具有良好的特异性;EBV检测核酸灵敏度为10^-7 ng,LFV、MRBV、RVFV、CCHFV、MERSV检测灵敏度均为10^-6 ng。结论建立的可同时检测EVB等6种输入性烈性传染病病毒的方法具有较高的灵敏度和特异性,可用于相关输入性传染病疫情的应急检测。
Objective To establish a method of simultaneously detecting Ebola virus(EBV),Lassa fever virus(LFV),Rift Valley fever virus(RVFV),Marburg virus(MBV),Crimean-Congo hemorrhagic fever virus(CCHFV),and Middle East respiratory syndrome virus(MERSV)by combining multiplex PCR amplification and Luminex liquid chip techniques in order to provide a technical reserve for detection of the relevant severely infectious diseases.Methods A multiplex PCR amplification reaction system was established for each of the six related viruses,and the virus-specific nucleic acid probe for each was coupled with differently encoded fluorescent microspheres.The products of amplification with PCR and microspheres coupled with different nucleic acid probes were used.The mixture was subjected to a hybridization reaction to establish a method for liquid phase chip detection of the six viruses,and its specificity and sensitivity were verified.Results Specificity testing indicated that there was no cross-interference between different viruses,indicating that the methods of detecting the six severely infectious viruses established in this study have good specificity;sensitivity testing indicated a sensitivity of 10^-7 ng for EBV;the detection sensitivity was 10^-6 ng for the five other viruses(LFV,MRBV,RVFV,CCHFV,and MERSV).Conclusion The established method for simultaneously detecting six types of imported infectious diseases such as EVB has a high level of sensitivity and specificity and can be used for emergency detection of related infectious diseases.
作者
戴莹
雷亚克
岳苗苗
胡兵
DAI Ying;LEI Ya-ke;YUE Miao-miao;HU Bing(Institute of Health Inspection and Testing,Disease Control and Prevention Center of Hubei Province,Wuhan,China 430079)
出处
《中国病原生物学杂志》
CSCD
北大核心
2019年第11期1303-1307,共5页
Journal of Pathogen Biology
关键词
多重PCR
液相芯片
输入性烈性传染病
检测
Multiplex PCR
Luminex liquid chip techniques
severely infectious diseases
detection
