流式细胞技术检测肾脏移植受者pp65抗原血症的意义  

Determination of pp65 antigen in renal transplant recipients by flow cytometry

作　　者：白剑[1] 石炳毅[1] 肖漓[1] 唐萌[1] 裴向克[1] 刘有[1] 张俊[2] 陈娟 

机构地区：[1]解放军总医院附属二院器官移植中心,北京市100091 [2]大连医科大学,辽宁瘩大连市116044 [3]解放军92538部队医院,辽宁省大连市116047

出　　处：《中国组织工程研究与临床康复》2008年第B12期10441-10444,共4页Journal of Clinical Rehabilitative Tissue Engineering Research

摘　　要：背景:临床对pp65 抗原的检测主要采用免疫荧光技术,但是该方法步骤繁琐,需要对标本进行人工计数,在检测过程中受人为的影响因素较多。目的:探讨使用简便的流式细胞技术检测人巨细胞病毒 pp65 抗原血症的可行性及其特点。设计、时间及地点:对比观察实验,于2007-09/2008-03 在解放军总医院附属二院器官移植中心完成。材料:肾脏移植受者外周血标本145份。方法:分离人外周血白细胞,分别采用免疫荧光技术与流式细胞技术对人巨细胞病毒 pp65 抗原检测。主要观察指标:流式细胞仪在激发波长为 488 nm 处的吸收峰判断阳性检出。结果:对肾脏移植受者进行免疫荧光技术检测,在 15 例受者的 25 份标本中检出阳性,检出的特异性为 76.0%;对肾脏移植受者进行流式细胞技术检测,在 5 例受者的 11 份标本中检出阳性,检出的特异性为 54.5%。使用 Fisher 确切概率法对两种不同检测方法检出的效果进行检验,结果两种检测方法的检出效果差异无显著性意义(P > 0.05)。结论:免疫荧光技术和流式细胞技术两种 pp65 抗原检测方法的检出效果无差异,因此可以通过流式细胞技术对 pp65 抗原进行检测,并且结果更加客观。BACKGROUND： Clinical determination of pp65 antigen relies on immunofluorescence assay, but this method is complicated and the results will be affected by many factors. OBJECTIVE： To investigate detecting feasibilitiy and characteristics of human cytomegalovirus pp65 antigen by flow cytometry. DESIGN, TIME AND SETTING： A controlled observation trial was performed in the Organ Transplantation Center, the Second Affiliated Hospital of Chinese PLA General Hospital （Beijing, China） from September 2007 to March 2008. MATERIALS： A total of 145 peripheral blood samples were collected from kidney transplantation recipients. METHODS： Human peripheral blood leucocytes were isolated. The human cytomegalovirus pp65 antigen was tested with flow cytometry and immunofluorescence assay. MAIN OUTCOME MEASURES： The absorption peak at excitation wave length of 488 nm was detected using flow cytometry to judge the masculine. RESULTS： The positive result of cytomegalovirus pp65 antigen in 25 samples from 15 kidney transplantation recipients was tested with immunofluorescence assay, and the findings indicated that the specificity was 76.0%. Meantime, the positive result of cytomegalovirus pp65 antigen in 11 samples from 5 kidney transplantation recipients was tested with flow cytometry, and the findings indicated that the specificity was 54.5%. There were no obvious difference of immunofluorescence detection and flow cytometry detection by Fisher＇s exact test （P 〉 0.05）. CONCLUSION： There are no obvious differences of immunofluorescence detection and flow cytometry detection, so flow cytometry could be used in cytomegalovirus pp65 antigen detection, and it is a more objective method.

关 键 词：巨细胞病毒 PP65抗原 流式细胞技术 免疫荧光技术 

分 类 号：R617[医药卫生—外科学]