不同荧光抗体剂量对HLA-B27的流式细胞结果的影响

张危; 杨国际; 陈卫文; 李宏伟; 李秀萍; 曾晓燕

不同荧光抗体剂量对HLA-B27的流式细胞结果的影响

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
4. 昆明医科大学继续教育学院

基金项目:

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);

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出版历程
- 收稿日期: 2017-04-22

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

Funds:

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

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.
- 强直性脊柱炎 /
- HLA-B27 /
- 抗体剂量 /
- 流式细胞术
Abstract: Objective To study the effects of different fluorescent antibody dosages of HLA-B27 on the flow cytometry results in peripheral blood lymphocyte of ankylosing spondylitis (AS) .Methods The expression intensities of peripheral blood lymphocyte HLA-B27 were detected in 22 cases of AS and 30 cases of healthy individual by the flow cytometry. The anti-human FITC-HLA-B27/PE-CD3 antibody kit was divided into five groups (2 μL, 4 μL, 8 μL, 15 μL, 30 μL) . BD company set the value of fluorescence intensity≥146 as a positive result. 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 than 8 μL, the results were positive, and the results were similar in 15 μL and 30 μL, while if the dosages of antibody were less than 8 μL, the results were false negative (P <0.05) . Conclusion The 8μL dosage of antibody was helpful to decrease the miss diagnosis of AS.Simultaneously, with the increase of the amount of antibodies, the results do not have a substantial increase.
- Ankylosing spondylitis /
- HLA-B27 /
- Antibody dosage /
- Flow cytometry

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参考文献(21)

[1]	[1]RUDWALEIT M, VAN DER HEIJDE D, LANDEW R, et al.The development of Assessment of Spondylo Arthritis international Society classification criteria for axial spondyloarthritis (part II) :validation and final selection[J].Ann Rheum Dis, 2009, 68 (6) :777-783.
[2]	[2]CHEN B, LI J, HE C, et al.Role of HLA-B27 in the pathogenesis of ankylosing spondylitis (Review) [J].Mol Med Rep, 2017, 15 (4) :1943-1951.
[3]	[3]QIAN Q, XU X, HE H, et al.Clinical patterns and characteristics of ankylosing spondylitis in China[J].Clin Rheumatol, 2017, 36 (7) :1561-1568.
[4]	[4]MOHAMMAD GHASEMI-RAD, HOSAM ATTAYA, EMAL LESHA, et al.Ankylosing spondylitis:A state of the art factual backbone[J].World J Radiol, 2015, 7 (9) :236-252.
[5]	[5]JULIN-SANTIAGO F, GARCA-GARC A C, GARC A-OLIVERA I, et al.Epidemiology of rheumatic diseases in Mixtec and Chontal indigenous communities in Mexico:A cross-sectional community-based study[J].Clin Rheumatol, 2016, 35 (Suppl 1) :35-42.
[6]	[6]SIEPER J, HOLBROOK T, BLACK C M, et al.Burden of illness associated with non-radiographic axial spondyloarthritis:A multiperspective European cross-sectional observational study[J].Clin Exp Rheumatol, 2016, 34 (6) :975-983.
[7]	[7]KOKO V, NDREPEPA A, SKNDERAJ S, et al.An epidemiological study on ankylosing spondylitis in southern Albania[J].Mater Sociomed, 2014, 26 (1) :26-29.
[8]	[8]ROHEKAR S, CHAN J, TSE S M, et al.2014 Update of the Canadian Rheumatology Association/spondyloarthritis research consortium of Canada treatment recommendations for the management of spondyloarthritis.Part I:principles of the management of spondyloarthritis in Canada[J].J Rheumatol, 2015, 42 (4) :654-664.
[9]	[9]MICHAEL M, WARD, ATUL DEODHAR, et al.American College of Rheumatology/Spondylitis Association of America/Spondyloarthritis Research and Treatment Network2015 Recommendations for the Treatment of Ankylosing Spondylitis and Nonradiographic Axial Spondyloarthritis Arthritis Rheumatol[J].Arthritis Rheumatol, 2016, 68 (2) :282-298.
[10]	[10]ISIDORO GONZLEZ-LVARO, CARMEN MARTNEZ FERNNDEZ, BENITO DORANTESCALDER N, et al.Spanish Rheumatology Society and Hospital Pharmacy Society Consensus on recommendations for biologics optimization in patients with rheumatoid arthritis, ankylosing spondylitis and psoriatic arthritis[J].Rheumatology (Oxford) , 2015, 54 (7) :1200-1209.
[11]	[11]JEFFREY R CURTIS, LESLIE R HARROLD, MARYAM M ASGARI, et al.Diagnostic Prevalence of Ankylosing Spondylitis Using Computerized Health Care Data, 1996 to 2009:Underrecognition in a US Health Care Setting[J].Perm J, 2016, 20 (4) :4-10.
[12]	[12]LI J, LIU Q, CHEN Y.Treatment patterns, complications, and direct medical costs associated with ankylosing spondylitisin Chinese urban patients:a retrospective claims dataset analysis[J].J Med Econ, 2017, 20 (1) :91-97.
[13]	[13]HONGTAO GUO, XUYAN NIU, YAN G U, et al.Differential Amino Acid, Carbohydrate and Lipid Metabolism Perpetuations Involved in a Subtype of Rheumatoid Arthritis with Chinese Medicine Cold Pattern[J].Int J Mol Sci, 2016, 17 (10) :1757.
[14]吴晓涛, 李传俊, 伍鑫, 等.磁共振成像对强直性脊柱炎早期骶髂关节炎诊断的临床应用价[J].昆明医科大学学报, 2017, 38 (3) :98-102.
[15]	[15]MAHMOUDI M, ASLANI S, NICKNAM M H, et al.New insights toward the pathogenesis of ankylosing spondylitis;genetic variations and epigenetic modifications[J].Mod Rheumatol, 2017, 27 (2) :198-209.
[16]	[16]HUI-CHUN Y U, MING-CHI L U, KUANG-YUNG HUANG, et al.Sulfasalazine Treatment Suppresses the Formation of HLA-B27 Heavy Chain Homodimer in Patients with Ankylosing Spondylitis[J].Int J Mol Sci, 2016, 17 (1) :46.
[17]	[18]CHENGGONG WANG, QIANDE LIAO, YIHE H U, et al.T lymphocyte subset imbalances in patients contribute to ankylosing spondylitis[J].Exp Ther Med, 2015, 9 (1) :250-256.
[18]	[17]XIA Q, WANG M, YANG X, et al.Autophagy-related IRGM genes confer susceptibility to ankylosing spondylitis in a Chinese female population:a case-control study[J].Genes Immun, 2017, 18 (1) :42-47.
[19]	[19]PAMELA B, WRIGHT, ANNE MCENTEGART, et al.Ankylosing spondylitis patients display altered dendritic cell and T cell populations that implicate pathogenic roles for the IL-23 cytokine axis and intestinal inflammation[J].Rheumatology (Oxford) , 2016, 55 (1) :120-132.
[20]	[20]WEI-CHIAO CHEN, JAMES CHENG-CHUNG W E I, HSING-FANG L U, et al.rs657075 (CSF2) Is Associated with the Disease Phenotype (BAS-G) of Ankylosing Spondylitis[J].Int J Mol Sci, 2017, 18 (1) :83.
[21]	[21]YONG L I, HONG-BO TANG, JING BIAN, et al.Genetic association between TNF-α-857 C/Tpolymorphism and ankylosing spondylitissusceptibility:evidence from a meta-analysis[J].Springerplus, 2016, 5 (1) :1930.

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