肺炎支原体快速检测方法的建立及应用评价  

Establishment and application evaluation of rapid detection method of Mycoplasma pneuomoniae

作　　者：张杰[1,2] 胡军[1,3] 刘杨[1,3] 李元[1,3] 

机构地区：[1]西安交通大学医学院,710049 [2]陕西省安康市中心医院输血科,725000 [3]陕西省人民医院中心实验室,西安710068

出　　处：《国际检验医学杂志》2016年第17期2379-2381,共3页International Journal of Laboratory Medicine

摘　　要：目的制备抗肺炎支原体(MP)的特异性单克隆抗体,利用单克隆抗体建立MP胶体金快速检测方法。方法采集急性呼吸道感染患儿咽拭子标本,分离培养并鉴定MP,纯化MP抗原,制备抗MP单抗,利用单抗建立MP胶体金检测试纸条。取疑似MP感染患者的咽拭子标本,分别使用荧光定量PCR法和胶体金快速检测法对其进行检测,观察并统计检查结果。结果成功分离培养出MP菌株,灭活并纯化出MP抗原,利用该抗原免疫小鼠,通过杂交瘤技术制备出抗MP单克隆抗体19株,从中选出效价高、特异性较好的两株单克隆抗体(MP-5和MP-19)作为原材料制备MP胶体金快速检测试纸条。胶体金试纸条最低检测限为20ng。分别采用胶体金试纸条和荧光定量PCR法对临床标本进行检测,结果显示,本研究制备的MP胶体金快速检测法的灵敏度为88.2%,特异度为82.6%。结论制备出抗MP的特异性单克隆抗体,已初步建立MP胶体金快速检测法,并为临床MP感染患者的快速诊断提供帮助。Objective To prepare the specific monoclonal antibodies（mAb）of Mycoplasma pneumonia（MP）and to establish a colloidal gold rapid detection method of MP by using mAb.Methods The nasopharyngeal swab samples were collected from children patients with acute respiratory tract infection,separated and cultured and performed the MP identification,MP antigen was purified,mAb of MP was prepared,then the MP colloidal gold test strip was established by using MP mAb.The throat swab sample was collected from the patients with suspected MP infection and detected by using the fluorescent quantitative PCR and colloidal gold test strip.The detection results were observed and statistically analyzed.Results MP strain was successfully isolated and inactivated,the MP antigen was purified.The mouse was immunized by using this antigen,19 strians of mAb against MP were prepared by using the hybridoma technique.Among them,2 strains of mAb with high titer and good specificity（MP-5 and MP-19）were selected as the raw materials for preparing the MP colloidal gold rapid test strip.The lowest detection limit was 20 ng.The clinical samples were detected by using the MP colloidal gold rapid test strip and fluorescent quantitative PCR.The results showed that the sensitivity of MP colloidal gold rapid test strip by using this established method was 88.2% and its specificity was 82.6%.Conclusion specific mAbs against MP is prepared and the colloidal gold rapid detection method is preliminarily established which provides the help for rapid diagnosis in the patients with MP infection.

关 键 词：肺炎支原体 单克隆抗体 胶体金免疫层析法 

分 类 号：R446.6[医药卫生—诊断学] R725.6[医药卫生—临床医学]