新型液相芯片复合检测5种鼠传病原体方法的建立  

Modern multiplexed suspension arrays for simultaneous detection of five Rodent-borne pathogens

作　　者：王旺[1] 杨宇[1] 王静[1] 赵婷婷[1] 

机构地区：[1]中国检验检疫科学研究院卫生检疫研究所,北京100123

出　　处：《中国国境卫生检疫杂志》2013年第2期73-76,共4页Chinese Journal of Frontier Health and Quarantine

基　　金：科技部国际合作项目(2010DFA34250)

摘　　要：目的建立一种能同时检测5种鼠传病原体:流行性出血热病毒(Epidemic hemorrhagic fever,EHF)、斑疹伤寒立克次体(Rickettsia typhi,TY1)、鼠疫耶尔森菌(Yersinia pestis,YP)、恙虫病立克次体(Rickettsia tsutsugamushi,RST)、钩端螺旋体(Leptospira,Lep)的液相芯片多重检测方法;方法针对5种病原的全基因组序列,分别设计特异性引物,并将上游引物5’端分别偶联一些特定的核酸片段作为标签(TAG),下游引物5’端标记生物素,PCR扩增后产物与微球进行杂交,最后利用液相芯片检测系统分析结果。结果建立的方法能同时完成对5种病原体Bio-plex100病原体及其中任何1种的检测,特异性好,灵敏度最低可达3 fg/test。结论通过利用偶联有TAG及生物素的引物对,成功建立了可同时检测5种鼠传病原的液相芯片检测方法,具有较高的灵敏度和特异性。该方法对于鼠传病原体的检测具有较好的应用前景。Objective To develop a new multiplexed X-TAG suspension arrays for simultaneous detection of rodent-borne pathogens including Epidemic hemorrhagic fever （EHF）, Rickettsia typhi（TY1）, Yersinia pestis（YP）, Rickettsia tsutsugamushi （RST） and Leptospira （Lep）. Methods Multi-PCR were designed with up-primers labeled by TAG on 5＇ end and down-primers labeled by biotin on 5＇ end separately, the PCR amplicons then hybridized to the coded magnetic microspheres coupled with anti-TAG, the conjugates were test and results were analyzed on Bio-plex 100. Results The method could be used to detect pathogens of any of these five types of rodent-borne pathogens and the sensitivity can reach 3 fg/test lowest. Conclusion The liquid biochip technology platform with the beads coupling with TAG was established which can simultaneously detect 5 types of rodent-borne pathogens, indicating a application potential for pathogen multiplex detection.

关 键 词：液相芯片 鼠传病原体 复合检测 标签 

分 类 号：R51[医药卫生—内科学]