Highly sensitive mutation quantification by high-dynamic-range capillary-array electrophoresis (HiDy CE).,Lab on a Chip

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Highly sensitive mutation quantification by high-dynamic-range capillary-array electrophoresis (HiDy CE).
Lab on a Chip ( IF 6.1 ) Pub Date : 2020-02-28 , DOI: 10.1039/c9lc00853e
Takashi Anazawa ₁ , Hiroko Matsunaga ₂ , Shuhei Yamamoto ₃ , Ryoji Inaba ₃

Affiliation

A simple and robust ultra-small four-color-fluorescence detection system was developed by integrating its components, namely, a four-capillary array, an injection-molded-plastic four-lens array, a four-dichroic-mirror array, and a CMOS sensor, as one device. The developed system was applied to a high-dynamic-range capillary-array electrophoresis (HiDy CE) to quantify a rare EGFR mutant (MT) of exon 19 deletion in a large excess of EGFR wild type (WT). Samples with serially diluted MT and constant-concentration WT were co-amplified by competitive PCR and subjected to HiDy CE. The MT peak in each electropherogram was then compared to the WT peak. As a result, MT was quantified with high-sensitivity (LOD of 0.004% MT/WT) and four-orders-of-magnitude dynamic range (0.01-100% MT/WT) by HiDy CE. Moreover, compared with existing methods, HiDy CE achieves higher speed, higher sample throughput, and lower consumable cost per sample. It has therefore great potential to be used in clinical practice.

中文翻译：

通过高动态范围毛细管阵列电泳（HiDy CE）进行高度敏感的突变定量。

通过集成其组件，即四毛细管阵列，注模塑料四透镜阵列，四分色镜阵列和光学组件，开发了一种简单而强大的超小型四色荧光检测系统。 CMOS传感器，作为一种设备。该开发的系统应用于高动态范围毛细管阵列电泳（HiDy CE），以量化在过量的EGFR野生型（WT）中稀有的外显子19缺失的罕见EGFR突变体（MT）。具有系列稀释的MT和恒定浓度WT的样品通过竞争性PCR共扩增，并进行HiDy CE。然后将每个电泳图中的MT峰与WT峰进行比较。结果，通过HiDy CE以高灵敏度（0.004％MT / WT的LOD）和四个数量级的动态范围（0.01-100％MT / WT）对MT进行了定量。而且，与现有方法相比，HiDy CE可实现更高的速度，更高的样品通量和更低的每个样品的消耗成本。因此，它在临床实践中具有很大的潜力。

更新日期：2020-03-19

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