ELISA检测肿瘤患者热休克蛋白90α的影响因素分析

Analysis of affecting factors in the detection of tumor patients heat shock protein 90α using ELISA

目的探讨影响ELISA定量检测肿瘤患者血浆中热休克蛋白90d（Hsp90a）的不同相关因素。方法实验采用比对研究，分别使用含EDTA—K2和EDTA．K3抗凝剂的真空采血管采集2012年3至7月在中国医学科学院肿瘤医院门诊就诊的72例肿瘤患者全血标本，进行Hsp90α定量检测。配制含不同浓度内源性干扰物质（溶血血红蛋白、乳糜等）的血浆样本，与正常样本同时检测Hsp90α的浓度。使用t检验分析各配对血浆样本中Hsp90α浓度值的差异。结果EDTA—K2抗凝管采集的血标本中Hsp9001浓度为（156．4±67．6）ng／ml，EDTA—K3抗凝管采集的血标本中浓度为（53．9±26．6）ng／ml，差异有统计学意义（t=10．68，P〈0．01）。乳糜浓度〉5％时血标本中Hsp90d浓度低于（34．3±2．0）ng／ml，正常血标本中浓度为（37．7±1．3）ng／ml，差异有统计学意义（t=3．96，P〈0．05）。不同浓度溶血的血标本中Hsp90仪浓度均大于（94．2±7．2）ng／ml，正常血标本中浓度为（67．0±4．4）ng／ml，差异有统计学意义（t=-9．17，P〈0．05）。类风湿因子（RF）浓度为25U／ml的血标本中浓度为（39．0±3．5）ng／ml，正常血标本中浓度为（35．3±1．7）ng／ml，差异有统计学意义（t=-2．37，P〈0．05）。黄疸浓度为4g／L时的血标本中浓度为（38．4±2．1）ng／ml，正常血标本中浓度为（35．7±1．4）ng／ml，差异有统计学意义（t=-2．97，P〈0．05）。结论外源性物质和内源性物质对ELISA检测血浆中Hsp90α的浓度均有干扰，在实验过程中应严格做好质量控制，尽量避免假阳性及假阴性结果的出现。

[ Abstract ] Objective To investigate the interference of the related factors on detection of heat shock protein 90α （Hsp90α） by ELISA. Methods Seventy-two blood samples were collected in Cancer Institute/ Hospital, Chinese Academy of Medical Sciences from March to July 2012, which collected simultaneously via various anticoagulants of EDTA-K2 and EDTA-K3. Different samples which were matched with normal plasma specimens, containing endogenous interfering substances （hemolytic hemoglobin, chyle） were prepared and analyzed. The differences of Hsp90α concentration in paired group were analyzed by t-test. Results There were statistical differences between EDTA-K2 group [ （ 156.4 ± 67.6） ng/ml ] and EDTA-K3 group [ （53.9 ± 26. 6） ng/ml, t = 10. 68, P 〈 0. 01 ]. Comparing the levels of Hsp90αin the chyle concentration lower than 5% groups[ （34. 3 ± 2. 0） ng/ml] with normal control group[ （37. 7 ± 1.3 ） ng/ml ], the value had significant difference （ t = 3.96, P 〈 0. 05 ）. The levels of Hsp90ct in hemolytic hemoglobin [ （94. 2 ± 7. 2） ng/ml were significantly higher than this of normal control [ （67.0 ± 4. 4） ng/ml, t = - 9. 17, P 〈 0. 05 ]. The levels of Hsp90ct in RF concentration 25 U/ml group [ （ 39.0 ± 3.5 ） ng/ml ] were higher than this of normal control[ （ 35.3 ± 1.7 ） ng/ml, t = - 2. 37, P 〈 0. 05 ]. The levels of Hsp90αin bilirubin concentration 4 g/L group [ （ 38.4 ± 2. 1 ） ng/ml ] were higher than this of normal control [ （ 35.7 ± 1.4） ng/ml, t = - 2. 97, P 〈 0. 05 ]. Conclusions Exogenous and endogenous substances would interfere the detection of Hsp90ct in tumor patients using ELISA method. Quality control should be implemented strictly during the experiments in order to avoid false positive and false negative results.