Germer Jeffrey J, Lins Michele M, Jensen Monica E, Harmsen W Scott, Ilstrup Duane M, Mitchell P Shawn, Cockerill Franklin R, Patel Robin
Division of Clinical Microbiology, Mayo Clinic, Rochester, Minnesota 55906, USA.
J Clin Microbiol. 2003 Aug;41(8):3503-8. doi: 10.1128/JCM.41.8.3503-3508.2003.
The COBAS AMPLICOR system has played a major role in the transition of molecular diagnostics from research to routine clinical laboratory use by automating the nucleic acid amplification and detection processes. However, sample preparation remains a labor-intensive portion of the procedure. In this study, we evaluated the performance of the COBAS AMPLICOR Hepatitis C Virus Test, version 2.0 (Roche Molecular Systems, Branchburg, N.J.) following manual hepatitis C virus (HCV) RNA extraction versus automated extraction with the MagNA Pure LC instrument (Roche Applied Science, Indianapolis, Ind.). Parallel replicate testing was performed with standard dilutions of 100, 75, 60, and 0 HCV IU/ml and 153 clinical specimens. An analytical sensitivity of 75 IU/ml was achieved with either the manual or the standard-volume (200 microl) automated extraction methodologies (25 of 26 [96.2%]; 95% confidence interval [95% CI], 80.4 to 99.9), whereas the clinical sensitivity and specificity were both 100% with either extraction method. A large-volume (1 ml) automated extraction method was also evaluated with standard dilutions of 40, 25, 10, and 0 IU/ml and the same 153 clinical specimens. The analytical sensitivity of the COBAS AMPLICOR assay with the large-volume extraction method was 25 HCV IU/ml (26 of 26 [100%]; 95% CI, 86.8 to 100), whereas the clinical sensitivity and specificity were both 100%. The MagNA Pure LC instrument is a versatile, labor-saving platform capable of integration with minimal modification of the existing assay procedure. The increased sensitivity of the COBAS AMPLICOR Hepatitis C Virus Test, version 2.0 performed in conjunction with large-volume HCV RNA extraction may be important in HCV diagnostic testing as new therapeutic strategies evolve.
COBAS AMPLICOR系统通过自动化核酸扩增和检测过程，在分子诊断从研究向常规临床实验室应用的转变中发挥了重要作用。然而，样本制备仍是该过程中劳动强度较大的部分。在本研究中，我们评估了COBAS AMPLICOR丙型肝炎病毒检测2.0版（罗氏分子系统公司，新泽西州布兰奇堡）在手动提取丙型肝炎病毒（HCV）RNA与使用MagNA Pure LC仪器（罗氏应用科学公司，印第安纳州印第安纳波利斯）自动提取后的性能。对100、75、60和0 HCV IU/ml的标准稀释液以及153份临床标本进行了平行重复检测。手动提取方法或标准体积（200微升）自动提取方法的分析灵敏度均达到75 IU/ml（26份中的25份[96.2%]；95%置信区间[95%CI]，80.4至99.9），而两种提取方法的临床灵敏度和特异性均为100%。还对40、25、10和0 IU/ml的标准稀释液以及相同的153份临床标本评估了大体积（1毫升）自动提取方法。大体积提取方法的COBAS AMPLICOR检测分析灵敏度为25 HCV IU/ml（26份中的26份[100%]；95%CI，86.8至100），而临床灵敏度和特异性均为100%。MagNA Pure LC仪器是一个通用、省力的平台，只需对现有检测程序进行最小程度的修改即可实现整合。随着新治疗策略的发展，结合大体积HCV RNA提取进行的COBAS AMPLICOR丙型肝炎病毒检测2.0版灵敏度的提高在HCV诊断检测中可能很重要。
