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1、呋喃哇酮代谢物间接竞争酶联免疫检测方法的研究AB®SMB黄登宇山西大学生命科学学院山西大学食品药品快速检测中心山西省食品药品监督管理局建立了呋喃唑酮代谢物间接竞争酶联免疫检测法。用活化酯法将卵清蛋白和半抗原连接成为被原。对毡被原和单克隆抗体的稀释倍数，封闭液浓度、竞争时间、酶标二抗孵育时间和显色反应时间进行优化，同时通过灵敏度、特异性、精密度和准确度等指标进行方法学评价。结果显示最佳条件为:包被原和单克隆抗体稀释倍数分别为800倍和6400倍，封闭液是1%脱脂乳，竞争时间是60min，辣根过氧化物酶标记羊抗鼠二抗孵育时间是45min,显色时间是15min。通过建立的标准曲线计算得
2、到线性方程是Y=-0.253X+1.336(R2=0.995),1。50是2.015ng/mL,线性范围是0.131〜30.911叩/0^,空白猪肉添加冋收率是84.8%〜90.3%,批内和批间变异系数分别为3.3%〜4.1%和4.6%〜6.7%。此方法特异性强,准确性、精密度和重现性均良好，可用于动物源性食品中呋喃唑酮代谢物的快速筛查。关键词：兽药;呋喃哇酮;酶联免疫检测法;史晓亚，，研宂方向为食品安全快速检测。:1297230283@qq.com黄登宇，，副教授，研究方向为食品卫生检测。:729845737@qq.com2017-09-16Studyofindirectcom
3、petitiveEnzymeLinkedImmunosorbentAssayfordetectionofFurazolidoneMetaboliteodSHIXiao-YaGAOLi—XiaHUANGDeng-YuCollegeofLifeScience，ShanxiUniversity;TheFoodandDrugSafetyRapidInspectionCenter，ShanxiUniversity;Abstract：Anindirectcompetitiveenzymelinkedimmunosorbentassay(ic-ELTSA)wasbaesdonmonoclona
4、lantibodywasdevelopedtodeterminefurazolidonemetabolite.Made3-{[(4-carboxyphcnyl)mcthylcne]amino}-2-oxazolidinone,CPAOZconjugatedwithOvalbumin,OVAbyusingN-hyduoxysuccinimideestermethodtosynthesizeCPAOZ-OVA.Thentheinfluenceofseveralphysicochemicalfactors(thedilutionratiosofCPAOZ-OVAandmonoclona
5、lantibody,blockingsolution,competitivereactiontime,incubationtimcforHRP-IgG，chromogenicreactiontime)ofthemethodwereoptimizedandthesensitivity,specificity,accuracyandprecisionwereevaluated.Intheoptimizedsystem,thedilutionratiosofCPAOZ-OVAandmonoclonalantibodywere800-foldand6400-foldrespectivel
6、y;blockingsolutionwas1%skimmedmilk;competitivereactiontimewas60min;incubationtimeforHRP-IgGwas45min;chromogenicreactiontime15min.Astandardcurvewasestablished,andthelinearequationforELISAwasY二-0.253X+l.336(R2二0.995)，IC5。was2.015ng/mL，thelinearityrangewas0.131〜30.911ng/mL,therecoveryrateinnegat
7、iveporksampleswasbetween84.8%~90.3%.Theintra—assayandinter-assaycoefficientsofvariationwere3.3%〜4.1%and4.6%〜6.7%respectively.Themethodwithdemonstratedexcellentspecificity,accuracy,precisionandreproducibilitycanprovidethebasisforrapiddetection
