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1、·70·匡堂生堂Q笙月28第l期JMedPostgra，Vo1．28，No．1，January，2015論著(技術創(chuàng)新)胃泌素釋放肽前體雙抗體夾心酶聯(lián)免疫吸附法的建立及其應用楚振宇，周小林，薛振偉，崔梅萍，藺蘇琴，李瑞華【摘要】目的胃泌素釋放肽前體(pro—gastrinreleasingpeptide，PGRP)作為小細胞肺癌腫瘤標志物的應用價值已成為近年來的研究熱點，文中建立新型雙抗體夾心酶聯(lián)免疫吸附法(enzyme—linkedimmunesorbentassay，ELISA)方法檢測患者血清PGRP抗原濃度。方法通過人工合成PGRP抗原決定簇對本實驗室自行研制的單克隆抗體進行篩選；采用
2、改良過碘酸鈉法對篩選到的單克隆抗體進行辣根過氧化物酶標記，建立PGRP雙抗體夾心酶聯(lián)免疫吸附檢測法；并與IBL公司PGRP試劑盒對比檢測結果。結果新型ELISA檢測標準抗原濃度范圍為33—1．7×10pg／mL，檢測小細胞肺癌患者血清PGRP濃度靈敏度100％，特異度98．4％，IBL試劑盒檢測靈敏度100％，特異度92．2％。結論新型ELISA方法可用于小細胞肺癌患者血清PGRP濃度檢測。[關鍵詞】胃泌素釋放肽前體；辣根過氧化物酶；單克隆抗體；酶聯(lián)免疫吸附試驗；腫瘤【中圖分類號】R34[文獻標志碼】A[文章編號】1008_8199(2015)叭一0070-04Establishmentofd
3、oubleantibodysandwichELISAforpro—gastrinreleasingpeptideanditsapplicationCHUZhen—yu，ZHOUXiao．1in，XUEZhen．wei，CUIMei．ping，LINSu．qin，LIRui．hua(1．DivisionofRadiologyandEnviromentalMedicine，ChinaInstituteforRadiationProtection，Taiyuan030006，Shanxi，China；2．DepartmentofMedicalOncology，TaiyuanTumorHospital
4、，Taiyuan030006，Shanxi，China)[Abstract]ObjectiveThevalueofpro—gastrinreleasingpeptide(PGRP)whichisthetumormarkerofsmallcelllungcanc—erhasbecomeahottopicinrecentyears．Theresearchwastobuildanewenzyme—linkedimmunesorbentassay(ELISA)methodai—ruingatdetectingtheconcentrationofPGRPinpatientsserum．MethodsWe
5、utilizedsyntheticPGRPepitopesforthescreeningofthemonoclonalantibodies，labeledthescreenedmonoclonalantibodieswithhorseradishperoxidasebymodifiedsodiumiodidemethod，andthenestablisheddoubleantibodysandwichELISAwhichcouldbeusedtodetecttheserumconcentrationsofPGRPincancerpa—tients．ResultsWesuccessfullysc
6、reenedEl2mAbwhichcouldbeservedasthecoatingantibodyandED1mAbasthelabeledanti—body．ThestandardantibodydensityrangeofnewELISAwas33pg／mL～1．7×10pg／mL．ThecomparisonexperimentsbetweenourmethodandthecommerciallyavailableELISAkitshowednosignificantdifference(P>0．05)．Thespecificityofourmethodwas50％，andthesens
7、itivitywas100％，whileIBLkitwas92．2％and100％respectively．ConclusionNewELISAcanbeusedtodetecttheserumPGRPconcentrationinpatientswithsmallcelllungcancer．[Keywords]Pro—gastrinreleasingpeptide；Horseradishper
