QIAGEN实时PCR与COBAS TaqMan检测血清HBV DNA预测慢性乙型肝炎肝组织病理状态的性能比较

摘要/Abstract

摘要： 目的比较QIAGEN实时PCR与COBAS TaqMan检测血清HBV DNA水平预测慢性乙型肝炎肝组织病理状态的性能。方法慢性乙型肝炎患者278例,其中HBeAg阳性和阴性分别为162例和116例。采用QIAGEN实时定量PCR和COBAS TaqMan系统检测血清HBV DNA。肝组织病理学诊断采用Scheuer评分系统,其中病理学分级和分期包括G0～G4和S0～S4。结果血清HBV DNA(QIAGEN)和HBV DNA(COBAS)在HBeAg阳性患者G2与G3,S1、S2、S3与S4之间的差异有统计学意义(P均<0.05);在HBeAg阴性患者G1与G2、G3,S1与S2、S3、S4之间的差异有统计学意义(P均<0.05)。血清HBV DNA(QIAGEN)与HBV DNA(COBAS)定量的不一致率,在HBeAg阳性患者G1-2、G3和S1-3、S4分别为4.24%(5/118)、9.09%(4/44)和3.68%(5/136)、7.69%(2/26),在HBeAg阴性患者G1、G2-3和S1、S2-4分别为6.02%(5/83)、3.03%(1/33)和3.29%(2/61)、3.64%(2/55)。血清HBV DNA(QIAGEN)和HBV DNA(COBAS)预测HBeAg阳性患者≥G3、≥S4的ROC曲线下面积分别为0.645和0.695、0.703和0.755,预测HBeAg阴性患者≥G2、≥S2的ROC曲线下面积分别为0.848和0.817、0.756和0.756。血清HBV DNA(QIAGEN)和HBV DNA(COBAS)预测HBeAg阳性患者≥S4的最佳截断值分别为≤3.784×10⁶ IU/mL和≤6.668×10⁷ IU/mL,对应的灵敏度、特异度分别为0.654和1.000、0.735和0.581;预测HBeAg阴性患者≥G2的最佳截断值分别为≥5.821×10³ IU/mL和≥9.311×10³ IU/mL,对应的灵敏度、特异度分别为0.970和0.909、0.614和0.651。结论血清HBV DNA预测肝组织病理状态的特点为预测HBeAg阴性患者≥G2的效能最大,并且血清HBV DNA(QIAGEN)与HBV DNA(COBAS)预测HBeAg阴性患者≥G2的性能有高度一致性。

关键词: HBV DNA定量, QIAGEN试剂, COBAS TaqMan系统, 慢性乙型肝炎, 肝组织, 病理学

Abstract: Objective To investigate and compare the performance of QIAGEN real-time PCR and COBAS TaqMan for serum hepatitis B virus (HBV) DNA quantitation in predicting the pathological state of liver tissue from chronic hepatitis B patients.Methods A total of 278 patients with chronic hepatitis B, including 162 HBeAg-positive and 116 HBeAg-negative ones, were enrolled in this study. Serum HBV DNA (QIAGEN) and HBV DNA (COBAS) were detected by real-time quantitative PCR and Roche TaqMan COBAS system, respectively. The Scheuer score system used for pathological diagnosis of liver tissue had 5 grades (G0-G4) and 5 stages (S0-S4), respectively. Results In HBeAg-positive patients, there were statistically significant differences of serum HBV DNA (QIAGEN) and HBV DNA (COBAS) between G2 and G3, S1 and S4, S2 and S4, S3 and S4 (all P<0.05), respectively; in HBeAg-negative patients, there were statistically significant differences between G1 and G2, G1 and G3, S1 and S2, S1 and S3, S1 and S4 (all P<0.05), respectively. Rates of disagreement between serum HBV DNA (QIAGEN) and HBV DNA (COBAS) quantitation of G1-2, G3, S1-3 and S4 in HBeAg-positive patients were 4.24% (5/118), 9.09% (4/44), 3.68%(5/136) and 7.69%(2/26), respectively; and the rates of G1, G2-3, S1 and S2-4 in HBeAg-negative patients were 6.02% (5/83), 3.03% (1/33), 3.29% (2/61) and 3.64% (2/55), respectively. Areas under the receiver operating characteristic (ROC) curves of serum HBV DNA (QIAGEN) and serum HBV DNA (COBAS) for predicting ≥G3 and S4 in HBeAg-positive patients were 0.645 and 0.695, 0.703 and 0.755, respectively; and the areas for predicting ≥G2 and ≥S2 in HBeAg-negative patients were 0.848 and 0.817, 0.756 and 0.756, respectively. Optimal cut-offs of serum HBV DNA (QIAGEN) and serum HBV DNA (COBAS) for predicting S4 in HBeAg-positive patients were ≤3.784×106 IU/mL and ≤6.668×10⁷ IU/mL, respectively; and the corresponding sensitivities and specificities were 0.654 and 1.000, 0.735 and 0.581, respectively; and the optimal cut-offs of serum HBV DNA (QIAGEN) and serum HBV DNA (COBAS) for predicting ≥G2 in HBeAg-negative patients were ≥5.821×10³ IU/mL and 9.311×10³ IU/mL, respectively; and the corresponding sensitivities and specificities were 0.970 and 0.909, 0.614 and 0.651, respectively. Conclusion The most efficiency of serum HBV DNA for predicting the pathological state of liver tissue is characterized by predicting ≥G2 in HBeAg negative patients, and there is a high consistance of the performance between serum HBV DNA (QIAGEN) and serum HBV DNA (COBAS) for predicting ≥G2 in HBeAg negative patients.

Key words: HBV DNA quantitation, QIAGEN reagent, Roche TaqMan COBAS system, Chronic hepatitis B, Liver tissue, Pathology

张占卿, 陆伟, 王平安, 王雁冰, 周新兰, 丁荣蓉, 李秀芬, 黄丹. QIAGEN实时PCR与COBAS TaqMan检测血清HBV DNA预测慢性乙型肝炎肝组织病理状态的性能比较[J]. 肝脏, 2016, 21(4): 241-247.

ZHANG Zhan-qing, LU Wei, WANG Ping-an, WANG Yan-bing, ZHOU Xin-lan, DING Rong-rong, LI Xiu-fen, HUANG Dan. Performance comparison between QIAGEN real-time PCR and COBAS TaqMan for serum HBV DNA quantitation in predicting the pathological state of liver tissue from chronic hepatitis B patients[J]. Chinese Hepatolgy, 2016, 21(4): 241-247.

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