1、中國(guó)農(nóng)業(yè)大學(xué)碩士學(xué)位論文利用IBV重組N蛋白建立ELISA抗體檢測(cè)試劑盒的研究姓名：王宏俊申請(qǐng)學(xué)位級(jí)別：碩士專業(yè)：傳染病與預(yù)防獸醫(yī)學(xué)指導(dǎo)教師：陳福勇2002.5.1AbstractThenucleoproteinproteinoftheGraystrainofinfectiousbronchitisvirus(IBV)ishighlyimmunogenicandcrossreactiveamongvariousdistinctserot

2、ypesRecombinantnucleoproteinexpressedinbacteriawithahistidinetagattheaminoterminushasbeenusedasantigenfordevelopingasantigenfordevelopingallassaytodetectIBVspecificantibodyThisfusionproteinwasproducedreadilyinbacteriaand

3、easilypurifiedwithanickelcolumnonwhichboundtothehistidinetagConditionswereoptimizedforusingthesepreparationsforanIBVspecificELISAItwasshownthattheoptimalconcentrationofrecombinantNproteinforcoatingofplatewas058Ug／m1theop

4、timalcoatingconditionofrecombinantNproteinforELISAwas4℃overnight，thedilutionofserumsamplewas1：100，theworkingconcentrationofHRP—labeledrabbitantichickenIgGwas1：1000，seromsamplefordetectingandHRPlabeledrabbitantichickenIgG

5、shouldbeincubatedat37“(2for30minrespectivelythesubstrateforELISAwasincubatedat3TCfor10minbeforereadingtheOD655FollowingthedeterminationofconditionsofELISA，thethresholdvalueofELISAWas014SerumsamplesagainstGrayCorm，Ark99，瓦

6、H，SE17B，Mass4twemdetectedfortheantibodytoIBVbyusingthedevelopedELISAItWaSfoundthattherewerecrossreactionl《tweentheantibodiesandtheRecombinantnucleoproteinMeanwhile，itwasconfirmedthattherewerenoCroSS—reactionbetweentheant

7、ibodiesagainstNDVEDS’76，Ecoil，IBDVAIVandSOonTheresultsrevealedthattheindireetELlSAbythepurifiedrecombinantnucleoproteinhadgoodspecificityforthedetectionofIBVantibodyinserumThedifferencevalueamongwellsinplateandamongplate

8、sforELISAWerebothlessthan10％whichshowedtheassayhadagoodreiterativity76serumsamplesfromchickenweredetectedfortheantibodytoIBVbyusingthedevelopedELISAandtheIDEXXIBVamibodytestkitsimultaneouslyItwasfoundthatthespecificityan