SYBR Green Ⅰ实时荧光定量PCR快速检测HLA-B27方法学的建立  

Establishment of SYBR Green Ⅰ Real-time PCR Method for Rapid Detection of HLA-B27

作　　者：王楠[1] 袁宏[1] 赵海英[2] 王丹丹[3] 

机构地区：[1]大连医科大学附属第一医院检验科,辽宁大连116023 [2]大连医科大学,辽宁大连116044 [3]辽宁省人民医院检验医学科,辽宁沈阳110016

出　　处：《医学与哲学（B）》2015年第11期67-69,83,共4页Medicine & Philosophy(B)

摘　　要：建立SYBR Green Ⅰ实时荧光定量PCR法快速检测HLA—B27，拟确定该法的阳性参考区间。强直性脊柱炎患者42例、类风湿性关节炎患者53例及健康体检者80例分别应用PCR—SSP法和SYBR Green Ⅰ实时荧光定量PCR法检测HLA—B27，并对两种方法进行比较。两种方法结果符合率为100％，初步拟定HLA—B27的阳性参考范围是：GAPDH的Ct在23．85-25．59，以99％百分位法建立HLA—B27阳性参考范围为CtB27〈21．54。SYBR Green Ⅰ实时荧光定量PCR法快捷、灵敏，适合于临床开展。To establish a method detecting HLA-B27 rapidly by real-time quantitative PCR with SYBR Green Ⅰ and to determine the positive reference interval. HLA-B27 DNA samples which came from 42 cases of clinical data confirmed AS patients, 53 cases of rheumatoid arthritis （RA） patients and 80 cases of healthy volunteers were detected respectively by PCR-SSP and SYBR Green Ⅰ real-time PCR, and the two methods were compared. In SYBR Green Ⅰ real-time PCR, the accordance rate of results was 100% by conventional PCR. When CtGAPDH was in the range of 23.85-25. 59, HLA-B27 positive reference interval was identified initially CtB27 〈21.54 by 99 % percentile method. SYBR Green Ⅰ real-time PCR was a rapid, sensitive method to detect HLA-B27 gene.

关 键 词：HLA-B27基因 强直性脊柱炎 SYBR Green Ⅰ 聚合酶链反应 

分 类 号：R446.9[医药卫生—诊断学]