DiDomenico N, Link H, Knobel R, Caratsch T, Weschler W, Loewy Z G, Rosenstraus M
Roche Molecular Systems, Branchburg, NJ 08876, USA.
Clin Chem. 1996 Dec;42(12):1915-23.
The COBAS AMPLICOR system automates amplification and detection of target nucleic acids, making diagnostic PCR routine for a variety of infectious diseases. The system contains a single thermal cycler with two independently regulated heating/cooling blocks, an incubator, a magnetic particle washer, a pipettor, and a photometer. Amplified products are captured on oligonucleotide-coated paramagnetic microparticles and detected with use of an avidin-horseradish peroxidase (HRP) conjugate. Concentrated solutions of amplicon or HRP were pipetted without detectable carryover. Amplified DNA was detected with an intraassay CV of < 4.5%; the combined intraassay CV for amplification and detection was < 15%. No cross-reactivity was observed when three different target nucleic acids were amplified in a single reaction and detected with three target-specific capture probes. The initial COBAS AMPLICOR menu includes qualitative tests for diagnosing infections with Chlamydia trachomatis, Neisseria gonorrhoeae, Mycobacterium tuberculosis, and hepatitis C virus. All tests include an optional Internal Control to provide assurance that specimens are successfully amplified and detected.
COBAS AMPLICOR系统可自动进行靶核酸的扩增和检测，使诊断性聚合酶链反应（PCR）成为多种传染病的常规检测方法。该系统包含一个带有两个独立调节加热/冷却模块的单一热循环仪、一个培养箱、一个磁性颗粒洗涤器、一个移液器和一个光度计。扩增产物被捕获在包被有寡核苷酸的顺磁性微粒上，并使用抗生物素蛋白-辣根过氧化物酶（HRP）偶联物进行检测。移取扩增子或HRP的浓缩溶液时未检测到残留。检测到的扩增DNA的批内变异系数（CV）<4.5%；扩增和检测的合并批内CV<15%。当在单个反应中扩增三种不同的靶核酸并用三种靶标特异性捕获探针进行检测时，未观察到交叉反应。最初的COBAS AMPLICOR检测项目包括用于诊断沙眼衣原体、淋病奈瑟菌、结核分枝杆菌和丙型肝炎病毒感染的定性检测。所有检测都包括一个可选的内部对照，以确保标本被成功扩增和检测。
