Caillat-Zucman S, Garchon H J, Costantino F, Cot S, Bach J F
INSERM U 25-Hôpital Necker, Paris, France.
Biotechniques. 1993 Sep;15(3):526-8, 530-1.
The ability to amplify specific genomic segments by the PCR has made the analysis of DNA polymorphism with oligonucleotidic probes a practical approach to HLA class II typing. However, to make this method more accurate and applicable to large-scale typing, technical and logistic limitations must be overcome and the risk of human errors must be reduced. To address this problem, we developed an automated procedure, using the Biomek 1000. The system consists of a workstation with a robotic arm and an instrument tablet, an electronic interface unit and an IBM PC. It was modified by the addition of a dot-blot apparatus. Automatic preparation of 96 samples for simultaneous DNA amplification is possible, together with PCR product dilution, distribution and dot-blotting. The robot prevents sample contamination, eliminates human errors and reduces operating costs by decreasing the working time by 50%. Thus, the system is suitable for routine analysis, and it also improves typing accuracy. Furthermore, this system can be adapted for a variety of new DNA-typing strategies that require an initial DNA amplification step.
通过聚合酶链反应（PCR）扩增特定基因组片段的能力，使得利用寡核苷酸探针分析DNA多态性成为进行人类白细胞抗原（HLA）Ⅱ类分型的一种实用方法。然而，要使该方法更加准确并适用于大规模分型，必须克服技术和后勤方面的限制，并降低人为误差的风险。为解决这一问题，我们利用Biomek 1000开发了一种自动化程序。该系统由一个带有机械臂和仪器操作平板的工作站、一个电子接口单元和一台IBM个人计算机组成。通过添加一个点杂交仪对其进行了改进。可以自动制备96个样本用于同时进行DNA扩增，同时还能进行PCR产物的稀释、分配和点杂交。该机器人可防止样本污染，消除人为误差，并通过将工作时间减少50%来降低运营成本。因此，该系统适用于常规分析，还能提高分型准确性。此外，该系统可适用于各种需要初始DNA扩增步骤的新DNA分型策略。
