不同荧光抗体剂量对HLA-B27的流式细胞结果的影响  

The Effects of Different Fluorescent Antibody Dosages of HLA-B27 on the Results of Flow Cytometry

作　　者：张危[1] 杨国际 陈卫文 李宏伟[3] 李秀萍 曾晓燕 ZHANG Wei;YANG Guo-ji;CHEN Wei-wen;LI Hong-wei;LI Xiu-ping;ZENG Xiao-yan(Dept. of Rheumatology,Qujing Affiliated Hospital of Kunming Medical University,Qujing Yunnan 655000;Dept. of Hepatobiliary Surgery,The Second People’s Hospital of Qujing,Qujing Yunnan 655000;Dept. of Biology laboratory,Qujing Affiliated Hospital of Kunming Medical University,Qujing Yunnan 655000;Continues Education Colledge of Kunming Medical University,Kunming Yunnan 650031,China)

机构地区：[1]昆明医科大学附属曲靖医院风湿免疫科 [2]曲靖市第二人民医院肝胆一科 [3]昆明医科大学附属曲靖医院生物实验室,云南曲靖655000 [4]昆明医科大学继续教育学院,云南昆明650031

出　　处：《昆明医科大学学报》2017年第8期98-101,共4页Journal of Kunming Medical University

基　　金：云南省科技厅-昆明医科大学应用基础研究联合专项基金资助项目(2014FZ060)

摘　　要：目的探讨不同剂量的流式细胞抗体对检测强直性脊柱炎(AS)患者外周血HLA-B27结果的影响.方法选择22例AS患者和30例正常人群,利用流式细胞仪检测外周血HLA-B27的表达情况.以美国BD公司设定的荧光强度界值≥146为阳性结果,抗人FITC-HLA-B27/PE-CD3抗体试剂盒分为5组(2μL、4μL、8μL、15μL、30μL).结果正常对照组,对应荧光强度结果为(55.97±6.44)、(59.90±7.15)、(67.97±7.62)、(81.60±6.63)、(91.97±5.77);确诊为AS的患者,荧光强度结果为(122.45±7.38)、(140.50±5.35)、(148.22±4.36)、(162.27±4.28)、(164.05±4.28);正常对照组结果随着抗体剂量的增高而上升,但均为阴性;AS患者8μL及以上抗体浓度时HLA-B27结果阳性,15μL、30μL抗体浓度结果相差不大,但抗体少于8μL后会出现假阴性,出现漏诊(P<0.05).结论应用8μL以上的HLA-B27抗体剂量,可以减少AS的漏诊;抗原抗体结合饱和后,随着抗体量的增加,结果没有明显变化.Objective To study the effects of different fluorescent antibody dosages of HLA-B27on the flowcytometry results in peripheral blood lymphocyte of ankylosing spondylitis(AS).Methods The expressionintensities of peripheral blood lymphocyte HLA-B27were detected in22cases of AS and30cases of healthyindividual by the flow cytometry.The anti-human FITC-HLA-B27/PE-CD3antibody kit was divided into five groups(2uL,4uL,8uL,15uL,30uL).BD company set the value of fluorescence intensity≥146as a positiveresult.Results The expression intensity in the healthy individuals were(55.97±6.44),(59.90±7.15),(67.97±7.62),(81.60±6.63),(91.97±5.77),while in AS were(122.45±7.38),(140.50±5.35),(148.22±4.36),(162.27±4.28),(164.05±4.28).The results rose along with the dosages of the antibody,the results in the healthy were negative.But in AS,if the dosages of antibody were more than8uL,the results werepositive,and the results were similar in15uL and30uL,while if the dosages of antibody were less than8uL,theresults were false negative(P<0.05).Conclusion The8μL dosage of antibody was helpful to decrease the missdiagnosis of AS.Simultaneously,with the increase of the amount of antibodies,the results do not have a substantial increase.

关 键 词：强直性脊柱炎 HLA-B27 抗体剂量 流式细胞术 

分 类 号：R593[医药卫生—内科学]