Advanced Liquid Logic, Research Triangle Park, NC, USA.
Clin Chem. 2011 Oct;57(10):1444-51. doi: 10.1373/clinchem.2011.163139. Epub 2011 Aug 22.
Newborn screening for lysosomal storage diseases (LSDs) has been gaining considerable interest owing to the availability of enzyme replacement therapies. We present a digital microfluidic platform to perform rapid, multiplexed enzymatic analysis of acid α-glucosidase (GAA) and acid α-galactosidase to screen for Pompe and Fabry disorders. The results were compared with those obtained using standard fluorometric methods.
We performed bench-based, fluorometric enzymatic analysis on 60 deidentified newborn dried blood spots (DBSs), plus 10 Pompe-affected and 11 Fabry-affected samples, at Duke Biochemical Genetics Laboratory using a 3-mm punch for each assay and an incubation time of 20 h. We used a digital microfluidic platform to automate fluorometric enzymatic assays at Advanced Liquid Logic Inc. using extract from a single punch for both assays, with an incubation time of 6 h. Assays were also performed with an incubation time of 1 h.
Assay results were generally comparable, although mean enzymatic activity for GAA using microfluidics was approximately 3 times higher than that obtained using bench-based methods, which could be attributed to higher substrate concentration. Clear separation was observed between the normal and affected samples at both 6- and 1-h incubation times using digital microfluidics.
A digital microfluidic platform compared favorably with a clinical reference laboratory to perform enzymatic analysis in DBSs for Pompe and Fabry disorders. This platform presents a new technology for a newborn screening laboratory to screen LSDs by fully automating all the liquid-handling operations in an inexpensive system, providing rapid results.
由于酶替代疗法的出现，新生儿溶酶体贮积症（LSD）的筛查引起了相当大的关注。我们提出了一种数字微流控平台，用于快速、多重酶分析酸性α-葡萄糖苷酶（GAA）和酸性α-半乳糖苷酶，以筛查庞贝病和法布里病。结果与使用标准荧光法获得的结果进行了比较。
我们在杜克生化遗传学实验室使用 3 毫米打孔器对 60 份未经识别的新生儿干血斑（DBS）以及 10 份庞贝病和 11 份法布里病样本进行了基于工作台的荧光酶分析，每种测定的孵育时间为 20 小时。我们使用数字微流控平台在 Advanced Liquid Logic Inc. 自动进行荧光酶分析，对于两种测定，使用单个打孔器提取的提取物，孵育时间为 6 小时。还进行了孵育时间为 1 小时的测定。
测定结果通常是可比的，尽管使用微流控的 GAA 酶活性平均值大约是使用基于工作台的方法获得的酶活性平均值的 3 倍，这可能归因于较高的底物浓度。使用数字微流控在 6 小时和 1 小时孵育时间，在正常和受影响的样本之间观察到明显的分离。
与临床参考实验室相比，数字微流控平台在 DBS 中进行庞贝病和法布里病的酶分析表现良好。该平台为新生儿筛查实验室提供了一种新技术，通过在廉价系统中完全自动化所有的液体处理操作，快速提供结果，从而筛选 LSD。
