乙型脑炎病毒IgM ELISA法测定.ppt
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1、IgM ELISA for Japanese encephalitis virus乙型脑炎病毒 IgM ELISA法测定,1,CHIK,SIN,DEN1-4,TBE(POW),SSH,EEE,WEE,VEE,MAY,DEN1-4,YF,BF,RR,SIN,MVE,DEN1-4,CHIK,SIN,YF,DEN1-4,TAH,SIN,TBE(POW),WN,(TAH,INK),LAC,SLE,POW,EEE,WEE,CTF,DEN1-4,EVE,EEE,VEE,WEE,MAY,DEN1-4,JE,(VEE),SLE,LAC,(VEE),DEN1-4,Most common arboviruses
2、World-wide 全球常见虫媒病毒分布,WN,WN,WN,WN,WN,SLE,SLE,2,uu,Flaviviruses in China 中国黄病毒Japanese encephalitis virus(JEV)乙型脑炎病毒Dengue viruses(DEN)登革热病毒Tick Borne encephalitis virus(TBE)(rare,near Russian border)蜱传脑炎病毒(TBE)(罕见,与俄罗斯边境处)There are 10 members of the JEV serocomplex,the main human pathogens of which
3、are JEV,West Nile(WNV),Murray Valley encephalitis virus(MVE)and St.Louis encephalitis virus(SLEV).Of these,only JEV is found in China乙脑病毒有10个成员,对人类致病的主要有乙脑病毒、西尼罗河病毒(WNV)、默里谷脑炎病毒(MVE)和圣路易斯脑炎病毒(SLEV)。其中,中国只发现乙型脑炎病毒,3,Flavivirus Virion Structure 黄病毒病毒体的结构,Virus particle 40-60 nmSingle-stranded positive
4、-sense RNA virus病毒颗粒4060nm单链正股RNA病毒,4,The Cross-Reactivity Problem 交叉反应问题,Human infections by flaviviruses elicitimmune responses producing 人感染黄病毒产生免疫反应,virus species-specific antibodies 病毒种特异性抗体 flavivirus cross-reactive antibodies 黄病毒交叉反应抗体,5,E Glycoprotein Homodimer E糖蛋白同型二聚体Top View 顶视图,Yellow r
5、esidues are domain II of E glycoprotein;purple are conserved regions among the flaviviruses黄色残基是E糖蛋白的结构域II;紫色为黄病毒的保守区域,6,Serologic cross-reactivity among the flaviviruses 黄病毒间血清交叉反应性Both IgM and IgG antibodies exhibit significant cross-reactivity among the flaviviruses.For instance,anti-DEN IgM will
6、 react with JE virus antigen,even though DEN is not in the JE serocomplex.黄病毒之间IgM和IgG抗体都有交叉反应。例如抗-登革热病毒IgM与乙脑病毒抗原反应,尽管登革热病毒不是乙脑病毒血清群的。IgM tends to be less cross-reactive than IgGThis can be useful in diagnosis of unexpected agents(eg when WNV entered the US it was first thought to be the endemic SL
7、E)However,cross-reactivity can produce false-positivesIgM通常比IgG交叉反应少些。可以用于非预期病原的诊断(如,当西尼罗河病毒进入美国时,最先认为是地方圣路易斯病毒脑炎)。然而,交叉反应可以产生假阳性。,7,Cross-reactivity continued交叉反应续IgM to DEN and TBE(flaviviruses in China)will cross-react in a JEV IgM test.登革热病毒和蜱传脑炎病毒(中国的黄病毒)IgM,在乙脑病毒IgM检测中可以发生交叉反应。TBE is rare in C
8、hina,therefore false-positives due to TBE are unlikely.蜱传脑炎在中国罕见,因此因蜱传脑炎病毒造成的假阳性可能性不大。DEN does not cause encephalitis but does cause occasional encephalopathy(disturbances in mental status),and therefore a specimen from a DEN patient could be submitted,and it is likely to test positive.登革热病毒不引起脑炎,但是
9、偶尔可以造成脑病(精神障碍),因此如果对登革热病人的标本进行检测,结果很可能阳性。,8,SHIPMENTS 运输,SERUM SPECIMENS:Acute and/or convalescent血清标本:急性期和/或恢复期Sent on wet,or dry ice or ambient temperature 在冰或干冰或常温环境运送CEREBROSPINAL FLUID 脑脊液Sent on wet or dry ice or ambient temperature 在冰或干冰或常温环境运送Store specimens at 4 C if intended for immediate
10、use;otherwise freeze at-20 C or below.如果准备立即检测,在4 C条件下保存标本;否则在-20 C或以下冻存,9,Information needed for serological testing and interpretation血清学检测和结果解释需要的资料,Dates of sample collection 样品采集日期Evidence of clinical encephalitis 临床脑炎证据Vaccination history 接种史Date of onset of symptoms症状出现日期,10,Safety安全性,Always
11、wear latex or nitrile gloves,safety glasses and a lab coat when handling specimens and during all stages of the test.处理样本和测试各个环节都要带乳胶和硅胶手套、安全眼镜和白大衣Use of a Biosafety cabinet(BSC)Class 2 Type A2 is recommended because acute specimens may contain infectious JE virus or other infectious agents.由于急性期标本可
12、能含传染性乙脑病毒或其它致病因子,建议用II类A2型生物安全柜Class 2 is a ventilated cabinet that provides personnel,product and environmental protection.It has a limited fixed inward air flow access opening,a ventilated downward HEPA filtered exhaust.II级生物安全柜有通风,可以提供人员、产品和环境保护。带有有限的、向内的固定气流,并使用了向下的HEPA过滤通气装置。Type 2 recirculates
13、 70%of the air II型有70的空气循环。Do not use laminar flow hoods intended for cell culture!细胞培养不要用层流罩。If a BSC is unavailable,care should be taken especially when using the undiluted specimens,and during wash steps.Bleach should be added to the discard pan to inactivate viruses.如果没有生物安全柜,操作要特别认真,特别是用未稀释的样品以
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