RF对ELISA检测血清HBeAg假阴性干扰的分析

doi:10.3969/j.issn.1673-8640.2017.09.005

摘要/Abstract

摘要：

目的证实类风湿因子（RF）对酶联免疫吸附试验（ELISA）检测乙型肝炎e抗原（HBeAg）假阴性干扰的存在。方法收集乙型肝炎表面抗原（HBsAg）、HBeAg均阴性而RF阳性新鲜血清样本,与HBeAg阳性对照品9∶1混合配成HBeAg阳性且RF阳性的试验样本,分别用不同的ELISA试剂检测样本的HBeAg。使用RF吸附乳胶对干扰样本进行预处理,比较预处理前后HBeAg水平的变化。结果试验中RF对3种不同厂商生产的HBeAg ELISA试剂均有不同程度的假阴性干扰。9例假阴性样本预处理后有7例转为阳性。结论 RF可影响ELISA检测HBeAg 的S/CO值,引起假阴性结果,样本预处理可以有效消除大部分RF的干扰。

关键词: 类风湿因子, 干扰, 酶联免疫吸附试验, 乙型肝炎e抗原

Abstract:

Objective To investigate whether rheumatoid factor（RF） can cause false negative interference on serum hepatitis B e antigen（HBeAg）by enzyme-linked immunosorbent assay（ELISA）.Methods Serum samples with negative hepatitis B surface antigen（HBsAg）,negative HBeAg and positive RF were collected and comprised by blending HBeAg-positive and RF-positive control substance at a ratio of 9∶1. HBeAg level was determined by different ELISA kits. RF adsorption latex was used to remove RF from samples,and the change of HBeAg level was compared before and after pre-treatment. Results The 3 kinds of HBeAg ELISA kits had false negative interference. There were 7 of 9 false negative samples transforming to positive after pre-treatment. Conclusions The S/CO of HBeAg determination by ELISA is susceptible to false negative interference from RF. The pre-treatment of samples is effective prior to the interpretation of determination results.

Key words: Rheumatoid factor, Interference, Enzyme-linked immunosorbent assay, Hepatitis B e antigen

中图分类号:

R446.62

谢良才, 范文, 徐磊, 许蓉, 刘天春. RF对ELISA检测血清HBeAg假阴性干扰的分析[J]. 检验医学, 2017, 32(9): 765-768.

XIE Liangcai, FAN Wen, XU Lei, XU Rong, LIU Tianchun. False negative interference of RF on serum HBeAg by ELISA[J]. Laboratory Medicine, 2017, 32(9): 765-768.

图/表 6

参考文献 15

[1]	LOK A S,MCMAHON B J.Chronic hepatitis B[J]. Hepatology,2001,34(6):1225-1241.
[2]	EMERSON J F,LAI K K Y.Endogenous antibody interferences in immunoassays[J]. Lab Medicine,2013,44(1):69-73.
[3]	TATE J,WARD G.Interferences in immunoassay[J]. Clin Biochem Rev,2004,25(2):105-120.
[4]	BARTELS E M,RIBEL-MADSEN S.Cytokine measurements and possible interference from heterophilic antibodies-problems and solutions experienced with rheumatoid factor[J]. Methods,2013,61(1):18-22.
[5]	FAN W,XU L,XIE L,et al.Negative interference by rheumatoid factor of plasma B-type natriuretic peptide in chemiluminescent microparticle immunoassays[J]. PLoS One,2014,9(8):e105304.
[6]	LI Y,WANG X,XU L,et al.Rapid identification of falsely elevated serum cardiac troponin I values in a stat laboratory[J]. Lab Med,2014,45(1):82-85.
[7]	BOSCATO L M,STUART M C.Incidence and specificity of interference in two-site immunoassays[J]. Clin Chem,1986,32(8):1491-1495.
[8]	ISMAIL A A.Interference from endogenous antibodies in automated immunoassays:what laboratorians need to know[J]. J Clin Pathol,2009,62(8):673-678.
[9]	JONES G R,GIANNOPOULOS P.A method for routine detection of heterophile antibody interference in the Abbott AxSYM hCG assay[J]. Pathology,2004,36(4):364-366.
[10]	TODD D J,KNOWLTON N,AMATO M,et al.Erroneous augmentation of multiplex assay measurements in patients with rheumatoid arthritis due to heterophilic binding by serum rheumatoid factor[J]. Arthritis Rheum,2011,63(4):894-903.
[11]	INGEGNOLI F,CASTELLI R,GUALTIEROTTI R.Rheumatoid factors:clinical applications[J]. Dis Markers,2013,35(6):727-734.
[12]	KRAGSTRUP T W,VORUP-JENSEN T,DELEURAN B,et al.A simple set of validation steps identifies and removes false results in a sandwich enzyme-linked immunosorbent assay caused by anti-animal IgG antibodies in plasma from arthritis patients[J]. Springerplus,2013,2(1):263.
[13]	ISMAIL A A,WALKER P L,CAWOOD M L,et al.Interference in immunoassay is an underestimated problem[J]. Ann Clin Biochem,2002,39(Pt 4):366-373.
[14]	OLSEN T G,HUBERT P R,NYCUM L M.Falsely elevated human chorionic gonadotropin leading to unnecessary therapy[J]. Obstet Gynecol,2001,98(5 Pt 1):843-845.
[15]	ROTMENSCH S,COLE L A.False diagnosis and needless therapy of presumed malignant disease in women with false-positive human chorionic gonadotropin concentrations[J]. Lancet,2000,355(9205):712-715.

组别	假阴性（S/CO值<1.00）			合计
组别	试剂Ⅰ	试剂Ⅱ	试剂Ⅲ	合计
弱阳性组	18/53	21/53	26/53	65/159
阳性组	4/49	6/49	7/49	17/147
强阳性组	0/55	0/55	0/55	0/165
合计	26/157	30/157	37/157

组别	假阴性（S/CO值<1.00）			合计
组别	试剂Ⅰ	试剂Ⅱ	试剂Ⅲ	合计
弱阳性组	18/53	21/53	26/53	65/159
阳性组	4/49	6/49	7/49	17/147
强阳性组	0/55	0/55	0/55	0/165
合计	26/157	30/157	37/157

组别	不同HBeAg水平的S/CO值
组别	水平1（43例）	水平2（24例）	水平3（25例）	水平4（41例）	水平5（24例）
试验组	1.17±0.80	1.49±0.91	8.09±3.48	13.47±6.57	22.43±9.40
对照组	1.63±0.15	2.40±0.17	8.34±0.56	19.60±1.03	29.80±2.37
P值	0.001	0.000	0.731	0.000	0.001

组别	不同HBeAg水平的S/CO值
组别	水平1（43例）	水平2（24例）	水平3（25例）	水平4（41例）	水平5（24例）
试验组	1.17±0.80	1.49±0.91	8.09±3.48	13.47±6.57	22.43±9.40
对照组	1.63±0.15	2.40±0.17	8.34±0.56	19.60±1.03	29.80±2.37
P值	0.001	0.000	0.731	0.000	0.001

样本	各稀释比例样本的HBeAg S/CO值
样本	原倍	1∶2	1∶4	1∶8	1∶16	1∶32
试验组
样本1	0.82	0.83	1.14	1.33	0.90	0.71
样本2	2.31	1.84	1.77	1.66	1.79	0.86
样本3	3.63	3.26	2.88	2.04	1.52	0.84
样本4	1.23	1.14	1.07	1.05	0.91	0.91
样本5	2.21	2.31	2.01	1.85	1.40	0.93
对照组
对照1	15.82	9.29	5.38	3.08	1.59	0.84
对照2	16.23	10.03	6.02	3.11	1.68	0.86