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1、第25卷第3期生物医学工程学杂志Vol.25No.32008年6月JournalofBiomedicalEngineeringJune20083肺炎衣原体特异性单克隆抗体的研制及应用1Δ212111王卫群钱利生史益军李雪萍白永怿徐坚余竹元1(复旦大学附属金山医院心内科,上海200540)2(复旦大学上海医学院病原生物学系微生物学教研室,上海200540)摘要采用肺炎衣原体(Chlamydiapneumoniae,Cpn)TW2183的纯化原体免疫BALB/C雄性小鼠,将小鼠脾脏细胞与SP2/0细胞融合,杂交细胞通过有限稀释法克隆,获得1株稳定分泌Cpn2单克隆抗体(Cpn2
2、McAb)的细胞株,单抗属IgG2b亚型及抗Cpn2MOMP抗体。自制Cpn2McAb与鹦鹉热衣原体原体有较弱的交叉反应;与沙眼衣原体原体无交叉反应,特性与进口Cpn2McAb一致。为了评估Cpn2McAb,用自制和进口Cpn2McAb同时检测外周血单核细胞标本454例,Cpn2Ag阳性率分别为5313%和5216%,Kappa值01714,两者有较高的一致性。结果提示,自制Cpn2McAb几乎和进口Cpn2McAb一样有较高的特异性和敏感性,可替代进口单抗用于临床Cpn感染的检测,有助于相关疾病的诊治。关键词肺炎衣原体特异性单克隆抗体微量免疫荧光外周血单核细胞中图分类号R
3、37413;R392111文献标识码A文章编号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
