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抗心磷脂抗体检测及其影响因素的研究 |
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裴瑾 米东辉 曲立科 杨翰仪
摘 要 目的：研究血清样品稀释度、温度、离子强度以及样品检测间 隔时间对血清抗心磷脂抗体(ACA)检测结果的影响。方法：采用酶联免疫吸附测定法(ELISA)检测ACA。结果：血清样品最佳稀释度为1∶50；37℃时光吸收值比22℃和25 ℃的光吸收值显著下降；15 mol/L NaCl可明显抑制ACA与抗原结合(P＜0.01)，抑制率 平均为45.4%；包被14 d以上的检测结果明显高于1 d和7 d的测定结果(P＜0.05)。结论：血清ACA检测结果受血清样品稀释度、温度、离子强度及包被时 间影响。
关键词 抗心磷脂抗体 酶联免疫吸附测定法 系统性红斑狼疮
中国图书分类号 R392.11
Studies on affected factors about antipholipid antibod ies determinations
PEI Jin,MI Dong-Hui,Qu Li-Ke et al.
(Basic College of Norman Bethune University of Medical Science,Changchun 130021)
Abstract Objective:Human sera ACA was detected by ELISA m ethod and some factors including the dilution rate of sera, blocking materials, te mperature,ionic strength and interval time between blocking and detection,which could probablly influence the detective level of sera ACA were investigated.[ WT5”HZ〗Methods:Human sera ACA was detected by ELISA.Results:10% NBS/PBS was more feasible than 1% bovine albu m in/PBS for blocking non-specific binding;the ACA titer of the same sample detec ted at 37℃ was lower than that of 22℃ and 25℃,the dilution rate 1∶50 for ser a was suitable for the detection; time internal between blocking and detection c ou ld influence ACA level when it exceeding 2 weeks;the blinding ability of Ag with Ab was signicantly inhibited when the NaCl inonic strength was higher than 1.5 mol/L.Conclusion:The detected results of human sera ACA influen c ed by the dilution rate of sera,blocking materials,temperature ionic strength a nd internal time between blocking and detection.
Key words ACA ELISA SLE
1983年Harris等应用放射免疫法测定SLE患者血清的ACA水平，该法具有较高的 敏感性［1］。以后经Loizou等改良得出与放射免疫法效果相同而更为方 便的酶联免 疫吸附试验(enzyme-linked immunosorbent assay,ELISA)法［2］。由于ELISA具有 高度敏感性 、可定量、可检测抗体类及亚类、易于标准化、快速简便等优点，目前已成为定量测定ACA 的标准方法［3］。尽管ACA的ELISA检测方法已经确立，但是不同的实验室所测得的 结果存在着一定的差异［4］。本实验在建立了用以检测抗心磷脂抗体的IgG、IgM、I gA三个亚类的ELISA法的基础上，对影响ACA检测结果的血清稀释度、温度等因素进行了深入 的研究，旨在使此检测系统更加完善，为进一步提高[1] [2] 下一页
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