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1、第25卷第3期生物醫(yī)學(xué)工程學(xué)雜志Vol.25No.32008年6月JournalofBiomedicalEngineeringJune20083肺炎衣原體特異性單克隆抗體的研制及應(yīng)用1Δ212111王衛(wèi)群錢(qián)利生史益軍李雪萍白永懌徐堅(jiān)余竹元1(復(fù)旦大學(xué)附屬金山醫(yī)院心內(nèi)科,上海200540)2(復(fù)旦大學(xué)上海醫(yī)學(xué)院病原生物學(xué)系微生物學(xué)教研室,上海200540)摘要采用肺炎衣原體(Chlamydiapneumoniae,Cpn)TW2183的純化原體免疫BALB/C雄性小鼠,將小鼠脾臟細(xì)胞與SP2/0細(xì)胞融合,雜交細(xì)胞通過(guò)有限稀釋法克隆,獲得1株穩(wěn)定分泌Cpn2單克隆抗體(Cpn2
2、McAb)的細(xì)胞株,單抗屬IgG2b亞型及抗Cpn2MOMP抗體。自制Cpn2McAb與鸚鵡熱衣原體原體有較弱的交叉反應(yīng);與沙眼衣原體原體無(wú)交叉反應(yīng),特性與進(jìn)口Cpn2McAb一致。為了評(píng)估Cpn2McAb,用自制和進(jìn)口Cpn2McAb同時(shí)檢測(cè)外周血單核細(xì)胞標(biāo)本454例,Cpn2Ag陽(yáng)性率分別為5313%和5216%,Kappa值01714,兩者有較高的一致性。結(jié)果提示,自制Cpn2McAb幾乎和進(jìn)口Cpn2McAb一樣有較高的特異性和敏感性,可替代進(jìn)口單抗用于臨床Cpn感染的檢測(cè),有助于相關(guān)疾病的診治。關(guān)鍵詞肺炎衣原體特異性單克隆抗體微量免疫熒光外周血單核細(xì)胞中圖分類號(hào)R
3、37413;R392111文獻(xiàn)標(biāo)識(shí)碼A文章編號(hào)100125515(2008)0320658204ProductionandApplicationofChlamydiaPneumoniae2specificMonoclonalAntibody1212111WangWeiqunQianLishengShiYijunLiXuepingBaiYongyiXuJianYuZhuyuan1(DepartmentofCardiology,JinshanHospital,FudanUniversity,Shanghai200540,China)2(DepartmentofMedicalMi
4、crobiologyandParasitology,ShanghaiMedicalCollege,FudanUniversity,Shanghai200540,China)AbstractThepurifiedelementarybodiesofC.pneumoniaeTW2183wereusedforimmunizationofmaleBALB/cmice,thespleencellsofthesemicewerefusedwithSP2/0cellsandthehybridcellswereclonedbylimitingdilution.Oneclonethatsec
5、retedtheC.pneumoniaemonoclonalantibody(Cpn2McAb)stablywasobtainedfinally.TheCpn2McAbbe2longedtoIgG2bclassandanti2Cpn2MOMP;theoutcomeofmicro2immunofluorescenceshoweditsweakcrossreactionwiththeC.psittacielementarybodybutithasnocrossreactionwithC.trachomaelementarybody.Ithasthesamespe2ciality
6、oftheimportedCpn2McAb.FortheevaluationofCpn2McAb,theperipheralbloodmononuclearcellspecimensof454patientsweredetectedbyself2madeCpn2McAbandimportedCpn2McAbatthesametime.ThepositiveratesofCpn2antigenwere5313%forself2madeCpn2McAband5216%forimportedCpn2McAb,showinghighconcordancebetweenthem(Ka
7、ppa=01714).Theresultsshowedthatself2madeCpn2McAbhasalmostthesamehighspecificityandsensitivityasimportedCpn2McAb,sotheself2madeCpn2McAbmayreplaceimportedCpn2McAbtodetectCpnspecificantigenandbehelpfultodiagnosingandtreatingtheclinicaldiseasesassociatedwithCpninf
