Elbeik Tarek, Markowitz Norman, Nassos Patricia, Kumar Uday, Beringer Scott, Haller Barbara, Ng Valerie
Department of Laboratory Medicine, University of California, San Francisco, CA, USA.
J Clin Microbiol. 2004 Jul;42(7):3120-7. doi: 10.1128/JCM.42.7.3120-3127.2004.
Branched DNA (bDNA) assays to quantify human immunodeficiency virus type 1 (HIV-1) and hepatitis C virus (HCV) consist of three distinct steps, including sample processing, hybridization, and detection, and utilize the System 340 platform for plate incubation and washing. Sample processing differs: HIV-1 from 1 ml of plasma is concentrated by high-speed centrifugation, whereas HCV plasma or serum samples are used without concentration. The first step of hybridization involves viral lysis at 63 degrees C: HIV-1 is performed in a heat block, whereas HCV is performed in System 340. The remaining hybridization and detection steps are similar for HIV-1 and HCV and executed on System 340. In the present study, the HIV-1 bDNA assay was adapted for viral lysis in the System 340 platform. The adaptation, test method 2, includes a 20-s vortex of concentrated viral pellet and lysis working solution, transfer of viral lysate to the 96-well capture plate, and transfer to System 340 programmed for HCV assay specifications. With test method 2, specificity and quantification were within assay specifications. HCV bDNA methodology remains unchanged. Hence, an HIV-1 and an HCV bDNA can be run simultaneously on System 340. With simultaneous testing, laboratories can run full plates, as well as combinations of full and partial plates. Also, simultaneous HIV-1 and HCV bDNA permits labor consolidation and improved workflow while maintaining multitasking and rapid patient result turnaround.
用于定量检测1型人类免疫缺陷病毒(HIV-1)和丙型肝炎病毒(HCV)的分支DNA(bDNA)检测法包括三个不同步骤,即样本处理、杂交和检测,并利用340系统平台进行平板孵育和洗涤。样本处理有所不同:1毫升血浆中的HIV-1通过高速离心进行浓缩,而HCV血浆或血清样本则无需浓缩直接使用。杂交的第一步是在63摄氏度下进行病毒裂解:HIV-1在热块中进行,而HCV在340系统中进行。HIV-1和HCV的其余杂交和检测步骤相似,并在340系统上执行。在本研究中,HIV-1 bDNA检测法被调整为在340系统平台上进行病毒裂解。这种调整后的测试方法2包括将浓缩病毒沉淀与裂解工作溶液涡旋20秒,将病毒裂解物转移至96孔捕获板,然后转移至按照HCV检测规格编程的340系统。采用测试方法2时,特异性和定量结果均在检测规格范围内。HCV bDNA方法保持不变。因此,HIV-1和HCV bDNA可以在340系统上同时进行检测。通过同时检测,实验室可以检测整板样本,也可以检测整板与部分板样本的组合。此外,同时检测HIV-1和HCV bDNA可实现人力整合并改善工作流程,同时保持多任务处理能力并加快患者结果反馈速度。