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时间:2017-11-13
《医药学临床医学毕业论文 肺炎衣原体抗体的制备及应用》由会员上传分享,免费在线阅读,更多相关内容在学术论文-天天文库。
1、湖南师范大学本科毕业论文考籍号:XXXXXXXXX姓名:XXX专业:医药学临床医学论文题目:肺炎衣原体抗体的制备及应用指导老师:XXX二〇一一年十二月十日作者:王卫群张瑞华余竹元【摘要】目的肺炎衣原体抗体(Cpn-PcAb)的制备及应用。方法用10μlCpn菌株TW-183(2×108ifu)与300μl福氏完全佐剂混匀后皮下多点注射新西兰白兔。双向琼脂扩散法鉴定血清抗体的效价为1∶64,用硫酸铵盐析法、DEAE-Sephadex-50层析法纯化免疫球蛋白(IgG),然后用异硫氰酸荧光素(FITG)标记IgG。用Cpn-PcAb与美国进
2、口Cpn-单抗(Cpn-McAb)同时检测病人外周血标本294例。另外,分别从Cpn-PcAb检测出Cpn特异性抗原阳性的外周血单核细胞(PBMC)标本(A组)中和对照组(B组)中随机各挑选20例,用PCR检测PBMC中的Cpn-DNA。结果Cpn-PcAb检测的阳性率为45.9%(135/294),Cpn-McAb检测的阳性率为43.5%(128/294),两种抗体同时阳性的标本有110例,Kappa值是07,根据其判断标准提示两者有较高的一致性。A、B两组Cpn-DNA阳性标本分别为17例和3例。结论Cpn抗体几乎和Cpn单抗一样
3、有较高的特异性和敏感性,可能可以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】肺炎衣原体多克隆抗体肺炎衣原体单克隆抗体微量免疫荧光外周血单核细胞Productionandapplicationofpolyclonalantibodytochlamydiapneumoniae【Abstract】ObjectiveProductionandapplicationofpolyclonalantibodytochlamydiapneumoniae(Cpn).MethodsTheNewZealandwhiterabbitw
4、assubcutaneouslyinjectedwith10μlCpnstrainTW-183(2×108ifu)and300μlcompleteFreundadjuvant.Theserumantibodytiterof1∶64wasidentifiedbytwodimensionalimmunodiffusiontest.IgGwasextractedfromserumandpurifiedbyammoniumsulphatesaltingoutandDEAE-SephadexA-50chromatography,thenwasla
5、beledbyfluoresceinisothiacyanate.InordertoevaluteCpn-PcAb,thebloodspecimensof294patientsweredeectedbyCpn-PcAbandCpnmonoclonalantibody(Cpn-PcAb)fromUSAatthesametime.Ontheotherhand,werandomlyselected20peripheralbloodmononuclearcell(PBMC)specimenswithCpnantigenpositive(grou
6、pA)and20oneswithCpnantigennegative(groupB)byCpn-PcAbdetection,thendetectedCpn-DNAinPBMCbyPCR.ResultsThepositiveratewas45.9%(135/294)forCpn-PcAband43.5%(128/294)forCpn-McAb,and110caseshavethesamepositiveforthem,showingthehighconcordancebetweenthem(Kappa=0.7).Thespecimenso
7、fpositiveCpn-DNAwerein17and3respectivelybetweengroupAandgroupB.ConclusionTheCpn-PcAbhasalmostthesamehighspecificityandsensitivityasCpn-McAb.TheCpn-PcAbmaybereplaceCpn-McAbtodetecttheCpnspecificantigenandbehelpfultodiagnoseandtreattheclinicaldiseasesassociatedwithCpninfec
8、tion.【Keywords】polyclonalantibodytochlamydiapneumoniae(Cpn-PcAb)monoclonalantibodytoCpn(Cpn-McAb)micro-
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