2种HLA-B27检测方法在强直性脊柱炎诊断中的比较  

Comparison of two kinds of HLA-B27 detection method in diagnosis of ankylosing spondilitis

作　　者：侯伟[1] 许晓东[1] 

机构地区：[1]卫生部北京医院血液科,北京100730

出　　处：《国际检验医学杂志》2014年第24期3398-3400,共3页International Journal of Laboratory Medicine

摘　　要：目的通过比较2种中国药监局(CFDA)批准的人类白细胞抗原(HLA)-B27体外诊断试剂盒,探讨荧光聚合酶链式反应(PCR)体外诊断试剂盒在HLA-B27检测中的临床应用价值。方法收集该院573例临床血液样本,分别采用基于荧光PCR技术和流式细胞法的2种上市获批试剂盒进行HLA-B27检测,对于2种方法检测结果不一致的样本采用PCR测序方法进行确认,同时随机抽取5%的荧光PCR法检测结果为阳性的样本进行复验。结果 573例样本,采用流式细胞法试剂盒检测出阳性191例,阴性382例;相同样本使用荧光PCR试剂盒检测HLA-B27基因阳性共194例,阴性379例。最终结果以流式细胞法作为参照,2种试剂盒阳性符合率为96.33%(184/191),阴性符合率为94.76%(362/382),27例样本不符(占总样本数的4.71%),2种方法的总符合率为95.29%[(184+362)/573],Kappa值为0.896,(P=0.02);四格表资料卡方检验P=0.021,2种方法检测结果一致性较高,但检测结果存在差异。复测标本中(包括27例检测结果不一致标本)HLA-B27基因测序结果与荧光PCR试剂盒检测判断结果一致。结论相比临床HLA-B27检测的权威方法流式细胞方法,荧光PCR法试剂盒在保证较高结果一致性的基础上,在HLA-B27检测上可能具有更为准确的判断能力,且相比标本制备和操作更为简单,具有较强的临床应用价值和前景。Objective To investigate the clinical application value of fluorescence PCR in vitro diagnosis（IVD）reagent kits in the HLA-B27 detection by comparing 2 kinds of HLA-B27 IVD reagent kit approved by CFDA.Methods A total of 573 clinical blood samples were collected and detected for HLA-B27 by the approved reagent kits based on the fluorescence PCR technique and the flow cytometry.The samples with inconsistent testing results by the two kits were further confirmed by the PCR sequencing.At the same time,about 5% samples of the positive results detected by the fluorescence PCR method were extracted for conducting the re-testing.Results Among 573 samples,191 samples were HLA-B27 positive and 382 cases were HLA-B27 negative by flow cytome-try;the same samples had 194 cases of HLA-B27 positive and 379 cases of HLA-B27 negative by real-time PCR.With flow cytome-try as reference of the final results,the positive coincidence rate of the two kinds of kit was 96.33%（184/191）,the negative coinci-dence rate was 94.76%（362/382）,27 samples had inconsistent results from the two kinds of assay（accounting for 4.71% of the to-tal number of samples）,the total coincidence rate was 95.29% [（184＋362）/573],the Kappa value was 0.896（P =0.02）;the chi-square test P =0.021,the two kinds of testing method had the high consistency,but the differences existed in the testing results. The re-testing results by PCR sequencing（including 27 samples with inconsistent results by two kinds of kit）were entirely consist-ent with the fluorescence PCR testing results.Conclusion Compared with the authority method flow cytometry for HLA-B27 tes-ting in clinic,the fluorescence PCR kit may present more accurate judging ability for the HLA-B27 testing on the basis of ensuring the higher consistency of the testing results,is easier compared with the sample preparation and operating procedures,and has the stronger clinical application value and prospects s.

关 键 词：流式细胞 荧光聚合酶链反应 强直性脊柱炎 

分 类 号：R593.23[医药卫生—内科学]