Clinical Application Efficiency Of Flow Cytometry In Detecting HLA-B27 Antigen Expression

Objective To explore the role of flow cytometry in the diagnosis and differential diagnosis of ankylosing spondylitis by detecting the expression of HLA-B27 antigen in human leukocytes,and the correlation between HLA-B27 antigen expression and lymphocyte subsets and Th1/Th2 immune balance.In addition,according to the results of genotyping HLA-B27 by PCR-SSP method,the results of flow cytometry and PCR-SSP method were compared to detect the expression level of HLA-B27,and then the efficacy of flow cytometry in clinical application was discussed.Methods 1.According to the preliminary screening of flow cytometry,the expression level of average fluorescence intensity of HLA-B27 was measured,and this study was conducted in groups.The analysis of lymphocyte subsets in human leukocytes and the expression of cytokines secreted by Th1/Th2 in T helper cells were detected by flow cytometry,and the correlation with HLA-B27 antigen expression was analyzed.2.Polymerase chain reaction-sequence-specific primer(PCR-SSP)was used to type HLA-B27 gene,and the results of the two methods were compared in consistency,so as to explore the clinical application efficiency of flow cytometry for HLA-B27 antigen expression.3.Statistical processing: SPSS22.0 statistical software is used for analysis.The non-normal distribution data in the measurement data is expressed by median(interquartile spacing),and the comparison between groups is analyzed by rank sum test.Pearson test was used for correlation analysis,P<0.05 was statistically significant.Results 1.Th1/Th2 immune imbalance exists in most patients with AS,especially the increase of IL-6 and TNF-α,and the average fluorescence intensity of HLA-B27 is significantly correlated with IL-6(P<0.05).The distribution of lymphocyte subsets in AS patients was abnormal,and the number of lymphocytes increased significantly,which was significantly related to the average fluorescence intensity of HLA-B27(P<0.05).2.Three subtypes of HLA-B27*04,HLA-B27*05/07 and HLA-B27*02 were detected by PCR-SSP genotyping,among which HLA-B27*05/07 was the most common.The consistency between flow cytometry and PCR-SSP was 95.7%.Conclusions 1.The distribution of lymphocyte subsets in AS patients is abnormal,the Th1/Th2 immune balance is destroyed,and the immune function of the body is disordered,which has a certain correlation with the positive expression of HLA-B27 antigen;2.The influence of "grey area" on the detection of HLA-B27 by flow cytometry is objective.The combination of PCR-SSP genotyping and flow cytometry can more accurately determine the antigen expression of HLA-B27 and the role of HLA-B27 in the diagnosis and differential diagnosis of clinically related diseases.