定量荧光PCR法验证流式细胞术测定人类白细胞抗原B27临界值标本的研究  

Quantitative fluorescent PCR to verify critical value specimens flow oytometry method determination of human leukocyte antigen B27 livability

作　　者：徐冉行[1] 李宇宁[1] 姚亚萍[1] 刘东海[1] 

机构地区：[1]宁波市第六医院检验科,浙江宁波315040

出　　处：《中华医院感染学杂志》2013年第9期2255-2257,共3页Chinese Journal of Nosocomiology

基　　金：宁波市自然科学基金资助项目(2010A610064)

摘　　要：目的观察用实时荧光定量聚合酶链反应(PCR)法来验证流式细胞术(FCM)检测人类白细胞抗原表达为临界值标本,评价其在强直性脊柱炎(AS)的临床诊治价值。方法分别对482例患者用流式细胞术检测人类白细胞抗原表达为阳性、临界值、阴性的样本及100名健康人群样本,以实时定量荧光PCR检测外周血HLA-B27DNA,确定样本的阳性率。结果 PCR法检测出阳性组调查282例阳性率100.00%、临界值组调查80例阳性率91.25%、阴性组调查120例阳性率为0.83%;健康对照组调查100名,阳性率3.00%。结论以定量荧光PCR检测>103为标准,FCM检测临界值组阳性率过高,差异有统计学意义,存在假阳性;阳性组和阴性组无明显差异,但阴性组有假阴性产生,定量PCR法对流式细胞术检测为临界值标本具有较高诊断价值,可防止假阳性和假阴性的产生,进一步提高AS诊断率和可靠性。OBJECTIVE To observe the critical value to verify the specimens flow cytometry（FCM） detection of human leukocyte antigen expression with the real-time fluorescence quantitative polymerase chain reaction（PCR） method and assess its value in the clinical diagnosis and treatment of ankylosing spondylitis（AS）.METHODS A total of 482 patients with flow cytometry detection of human leukocyte antigen expression were positive,the critical value,negative samples and 100 samples of healthy people,the peripheral blood of HLA-B27 DNA were detected by quantitative real-time fluorescence PCR to determine the positive rate of samples.RESULTS There were 282 cases tested in the positive group with the PCR with the positive rate of 100.00%,there were 80 cases tested in the critical value group with the positive rate of 91.25%,and there were 120 cases tested in the negative group with the positive rate of 0.83%.Of 100 cases investigated in the control group,the positive rate was 3.00%.CONCLUSION With the quantitative fluorescent PCR detection higher than 103 as the standard,the positive rate of FCM threshold is too high,the difference is statistically significant,and there is a false positive case;there is no significant difference between the negative group and the positive group,but there is false negative case in the negative group.The quantitative PCR has significant value in the diagnosis of convection cytometry detection of specimens of the critical value so as to prevent the false positive case and false negative case,increasing the diagnosis rate of AS and the reliability.

关 键 词：聚合酶链反应 强直性脊柱炎 流式细胞术 

分 类 号：R446[医药卫生—诊断学]