EB病毒急性感染的实验室血清学诊断方法研究

Research of Laboratory Serological Diagnostic Method for Acute EB Virus Infection

目的对化学发光免疫分析法（CLIA）测定EB病毒衣壳抗原Ig M（EBV-VCA Ig M）的结果进行方法学评价,并与酶联荧光分析法（ELFA）、酶联免疫吸附试验法（ELISA）诊断EB病毒（EBV）急性感染的效能进行比较。方法选取2011年5月—2012年10月北京大学人民医院不明原因发热患者142例,对受试者使用分离胶真空采血管抽取静脉血3 ml,分离血清,采用CLIA、ELFA和ELISA检测EBV-VCA Ig M,比较3种方法诊断EBV急性感染的价值;并对CLIA进行方法学评价。结果 CLIA低、中和高值（分别为25.8、81.8、184.4 U/ml）血清标本批内变异系数（CV）分别为5.7%、3.9%和2.6%,批间CV分别为11.4%、5.4%和4.2%。理论值与实测值间的回归方程为Y=0.090 4＋1.005 2X,相关系数r=0.998（P〈0.001）,回收率为95.3%~104.8%;当标本中血红蛋白水平达8 g/L,三酰甘油达30 000 mg/L时,均不干扰CLIA。CLIA诊断EBV急性感染的灵敏度为94.2%,特异度为91.1%,Youden＇s指数为0.853;ELFA诊断EBV急性感染的灵敏度为82.7%,特异度为88.9%,Youden＇s指数为0.716,;ELISA诊断EBV急性感染的灵敏度为80.8%,特异度为91.1%,Youden＇s指数为0.719。CLIA诊断EBV急性感染ROC曲线下面积（AUC）为0.962〔SE=0.021,95%CI（0.920,1.004）〕;ELFA诊断EBV急性感染的AUC为0.960〔SE=0.015,95%CI（0.911,0.990）〕;ELISA诊断EBV急性感染的AUC为0.882〔SE=0.031,95%CI（0.823,0.942）〕。ROC曲线显示,CLIA诊断EBV急性感染的最佳诊断界值为41 U/ml,灵敏度为96.2%,特异度为99.7%,Youden＇s指数为0.959。结论 CLIA是目前测定EBV-VCA Ig M较灵敏的定量检测方法,优于ELFA和ELISA,适用于临床EBV感染的早期诊断。

Objective To appraise the methodology of anti EBV- VCA Ig M assay by CLIA and compare its diagnostic effect of acute EBV virus infection with ELFA and ELISA. Methods 142 cases of origin- unknown fever in Peking University People＇s Hospital from May,2011 to October,2012 were selected as the research subjects. 3 ml of venous blood was drawn with separation gel vacuum collective tube and blood serum was separated for detecting EBV- VCA Ig M by CLIA, ELFA and ELISA. The diagnostic effects of the three methods were compared and methodology evaluation was then performed for the detection by CLIA. Results The coefficients of variation（ CV） of low,middle and high value of CLIA（ 25. 8,81. 8,184. 4 U / ml）in one batch inside were 5. 7%,3. 9% and 2. 6% respectively and in different batches separately were 11. 4%,5. 4% and 4. 2% respectively. The regression equation between expected values and measured values was Y = 0. 090 4 ＋ 1. 005 2X（ r =0. 998,P〈0. 001） and the recovery rate was 95. 3%- 104. 8%; hemoglobin level reaching 8 g / L and triglyceride reaching 30000 mg / L did not interfere with CLIA. The sensitivity of CLIA, ELFA and ELISA in diagnosing acute EBV infection were 94. 2%,82. 7% and 80. 8% separately, the specificity of the three were 91. 1%,88. 9% and 91. 1% separately and the Youden＇s index were 0. 853,0. 716 and 0. 719 separately. The AUC of CLIA for diagnosing acute EBV infection was 0. 962 〔SE= 0. 021,95% CI（ 0. 920,1. 004） 〕,the AUC of ELFA and ELISA were 0. 960 〔SE = 0. 015,95% CI（ 0. 911,0. 990） 〕and 0. 882 〔SE = 0. 031,95% CI（ 0. 823,0. 942） 〕 respectively. ROC curve showed that the best diagnostic threshold was 41U / ml,the sensitivity was 96. 2%, specificity was 99. 7% and Youden＇ s index was 0. 959. Conclusion CLIA is a more sensitive quantitative method for EB- VCA Ig M detection,and is superior to ELFA and ELISA. It is suitable for the early clinical diagnosis of acute EBV infection.