Jou C, Rhoads J, Bouma S, Ching S, Hoijer J, Schroeder-Poliak P, Zaun P, Smith S, Richards S, Caskey C T
Diagnostics Division, Abbott Laboratories, North Chicago, Illinois 60612, USA.
Hum Mutat. 1995;5(1):86-93. doi: 10.1002/humu.1380050112.
The purpose of this study is to demonstrate the value of a multiplex amplification and readout system. The validation was done using as a model system the detection of deletions in nine possible dystrophin exons: 4, 8, 12, 17, 19, 44, 45, 48, and 51. The amplification system was gap ligase chain reaction, adapted to amplify selected regions of multiple exons simultaneously. The amplified products were read out with an immunochromatographic methodology, adapted from that used in the Abbott product line commercialized under the name Test Pack Plus. In each amplification, the beta-globin gene was incorporated and served as a procedural control. The complete process takes < 3 hr from DNA sample to result. The procedure is therefore rapid and simple, as well as being potentially very cost effective. The combination of these two technologies is shown to be a useful tool for the determination of deletions in the nine exons of the dystrophin gene. The results of a 100-patient sample study showed concordance with cDNA and PCR in current use. Equivalent performance at two sites was shown.
本研究的目的是证明多重扩增和读出系统的价值。验证过程以检测九个可能的肌营养不良蛋白外显子（4、8、12、17、19、44、45、48和51）中的缺失作为模型系统来进行。扩增系统是缺口连接酶链反应，适用于同时扩增多个外显子的选定区域。扩增产物采用免疫层析方法读出，该方法改编自雅培产品线以Test Pack Plus为名商业化的产品所使用的方法。在每次扩增中，均掺入β-珠蛋白基因并用作程序对照。从DNA样本到得出结果，整个过程耗时不到3小时。因此，该程序快速、简单，而且可能极具成本效益。这两种技术的结合被证明是确定肌营养不良蛋白基因九个外显子中缺失情况的有用工具。一项对100名患者的样本研究结果表明，与目前使用的cDNA和PCR方法结果一致。在两个地点展示了同等性能。
