| Kitsize | 12x8 |
| Method | ELISA |
| Incubationtime | 1x1h,3x30min,1x15min |
| Standardrange | cut-offindex |
| Specimen/Volumes | 10µlserum |
| Substrate/isotope | TMB450nm |
| RegulatoryStatus: | EU:CE |
IBLInternationaloffersahighlysensitiveandspecificELISAtestintendedforthediagnosticwork-upofchikungunyafever,aninfectiousdiseasewhoseincidencehasbeenincreasingsteADIlyinrecentyears.Itscharacteristicsare:
- µ-capturetechnology
- Diagnosticsensitivityof>98%anddiagnosticspecificityof100%
- Indicatedforthedifferentialdiagnosisofchikungunyafeveranddenguefever
- Qualitativeresults(positive,negative,cut-offcontrol)
- Adaptabletoopen-systemELISAanalyzers
Chikungunyafeverisaviralfebrileillnesstransmittedbyseveralmosquitospecies.Itmayleadtoseverearthralgiathatmaypersistforweekstomonths.Nevertheless,chikungunyafeverisessentiallyself-limitingandnon-fatal.ChikungunyahashistoricallybeenlimitedtocountriesinAfrica,theIndiansubcontinentandSoutheastAsia.However,manytouristareasarenowalsoaffectedbyextensiveoutbreaksandincreasingincidence,whichishowthevirusmadeitswaytotheWesternHemisphere,resultinginimportedcasesinanumberofEuropeancountries.In2006,severalimportedchikungunyacaseswerediagnosedinNorthernItalyduetothepresenceofcompetentmosquitovectorsforthechikungunyavirus.
Laboratorydiagnosisisgenerallyaccomplishedbytestingseraumorplasmatodetectvirus-specificimmunoglobulin(IgM,IgG)bymeansofanimmunoassay.Sincethesymptomsandgeographicspreadofchikungunyaanddenguefeveroverlap,adifferentialdiagnosticwork-upisofutmostimportance,puttingspecialperformancedemandsupondiagnostictests.Moreover,duetotheabsenceofwidespreadSEROpositivityintheEuropeanpopulation,anypositiveantibodytestissUSPiciousforinfection.InadditiontotheChikungunyaIgMµ-captureELISA,IBLInternationaloffersanIgGcaptureELISA.Bothassaysboastoutstandingperformancedata.
EvaluationofdiagnosticperformanceoftheChikungunyaIgMµ-captureELISA
1.ExternalmethodcomparisonoftheChikungunyaIgMµ-captureELISAofferedbyIBLInternationalwithanin-houseimmunofluorescencetest(TheBernhardNochtInstituteforTropicalMedicine,Hamburg)
Thecomparisonof31positiveandnegativeserarevealedaconcordanceof96.8%.
| Immunofluorescence | |||
| positive | negative | ||
| IBLInternational | positive | 21 | 0 |
| negative | 1 | 9 | |
2.Analysisofselectedpositiveandnegativesera(DGHS,NewDelhi)usingtheIBLInternationalChikungunyaIgMµ-captureELISAassay
Theanalysisof88serarevealedanagreementof100%.
| DGHS | |||
| positive | negative | ||
| IBLInternational | positive | 52 | 0 |
| negative | 0 | 36 | |
ExcerptfromtheInstructionsforUse
ChikungunyavirusisanarthropodbornevirusofthegenusAlphavirus(familyTogaviridae).TheAlphavirusgenuscontainsatleast24distinctspecies.Thesearelipid-envelopedvirionswithadiameterof50to60nm.Alphavirusinfectionsareinitiatedbythebiteofaninfectedmosquito,whichresultsinthedepositionofvirusinsubcutaneousandpossiblycutaneoustissues.Afteranincubationperiodof1to12daystheChikungunyafeverdevelops.Chikungunyafever(Chikungunyameans“thatwhichbendsup”,inreferencetothecripplingmanifestationsofthedisease)isanacuteviralinfectioncharacterizedbyarapidtransitionfromastateofgoodhealthtoillnessthatincludesseverearthralgiaandfever.Temperaturerisesabruptlytoashighas40°Candisoftenaccompaniedbyshakingchills.Afterafewdays,fevermayabateandrecrudesce,givingrisetoa“saddleback”fevercurve.Arthralgiaispolyarticular,favoringthesmalljointsandsitesofpreviousinjuries,andismostintenseonarising.Patientstypicallyavoidmovementasmuchaspossible.Jointsmayswellwithoutsignificantfluidaccumulations.Thesesymptomsmaylastfrom1weektoseveralmonthsandareaccompaniedbymyalgia.Therashcharacteristicallyappearsonthefirstdayofillness,butonsetmaybedelayed.Itusuallyarisesasaflushoverthefaceandneck,whichevolvestoamaculopapularormacularformthatmaybepruritic.Thelatterlesionsappearonthetrunk,limbs,face,plamsandsoles,inthatorderoffrequency.Petechialskinlesionshavealsobeennoted.Headache,photophobia,retro-orbitralpain,sorethroatwithobjectivesignsofpharyngitis,nauseaandvomitingalsooccurinthissetting.Occasionally,howeverpersistentarthralgiaandpolyarthritis(lastingmonthsorevenyears)dooccur,sometimesinvolvingjointdestruction.Evenrarer,sequelaeincludeencephalitisandmeningoencephalitiswithhighlethalityrates.ThevirushasmajorimportanceinAfricaandAsia.From20%tomorethan90%ofthepopulationoftropicalandsubtropicalshowserologicevidenceofinfection.BecauseAedesmosquitoesareincreasinglyprevalentinNorthAfricaandSouthAmerica,wherethepopulationwouldbeuniformlysusceptibletoinfection,thepossibilityforepidemicsisevident.ChikungunyavirusinfectionsareimportedtocentralEuropemainlybytravellerstotropicalandsubtropicalcountries.Thepresenceofvirusresp.infectionmaybeidentifiedbySerology:DetectionofantibodiesbyIF,ELISA
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ELISA双抗体夹心法(enzyme linked immunosorbent assay——sandwich technique)的原理是将特异性抗体结合到固相载体上形成固相抗体，然后和待检血清中的相应抗原结合形成免疫复合物，洗涤后再加酶标记抗体，与免疫复合物中抗原结合形成酶标抗体-抗原-固相抗体复合物，加底物显色，判断抗原含量。
生物帮有相关介绍。编码RNA http://doc.bio1000.com/show-3399.html
请大家帮帮忙，第一步加完抗原后必须封闭吗
表面抗体是双抗原夹心法
E抗体和核心抗体是竞争抑制法或间接法
在这种测定方法中有3种必要的试剂：①固相的抗原或抗体（免疫吸附剂） ②酶标记的抗原或抗体（标记物）③酶作用的底物（显色剂）
测量时，抗原（抗体）先结合在固相载体上，但仍保留其免疫活性，然后加一种抗体（抗原）与酶结合成的偶联物（标记物），此偶联物仍保留其原免疫活性与酶活性，当偶联物与固相载体上的抗原（抗体）反应结合后，再加上酶的相应底物，即起催化水解或氧化还原反应而呈颜色。
其所生成的颜色深浅与欲测的抗原（抗体）含量成正比。 这种有色产物可用肉眼、光学显微镜、电子显微镜观察，也可以用分光光度计（酶标仪）加以测定。其方法简单，方便迅速，特异性强。向左转|向右转
2.加大辣根过氧化物酶标记的链霉亲和素；
1.标准曲线的标准品是否一定要梯度稀释，为什么？我试过非梯度稀释的，也可以达到线性R2=0.99.
2.我用了CurveExpert做标曲，自动搜索后发现有10种提供的方程，各种形式的，其中一个十分适合我的实验结果（LogisticModel），而其他的感觉又不适合，因为结果常常为负值。这又是为啥捏？
3.实验的酶标仪最大OD值可以测到4，如果我的测量结果在1.3，是否像其他人所说的＞1了就不准确了。
4.利用夹心法进行定量分析是否一定要使用线性方程？
不好意思啊，一下问了这么多问题，最近做了一个月的ELISA，完全摸不清头脑啊。谢谢各位了
