A microfluidic chip integrated with a microheater and a luminescent temperature sensor for rapid, spatial melting curve analysis was developed and applied for the screening of a breast cancer gene fragment. The method could detect genetic differences in around 3 minutes total for the whole procedure, which is much faster than established procedures. A microfabrication technique was developed to allow for bonding of a temperature sensing thin film and a Pt microheater with PDMS and the chips could be employed to generate and measure thermal gradients and the fluorescence intensity of stained DNA through multispectral optical imaging. The sensing layer consisting of poly(styrene-co-acrylonitrile) and a tris(1,10-phenanthroline)ruthenium(II) temperature probe was generated by blade coating on a glass substrate with an attached Pt microheater. Calibration of the temperature between 20 and 90 °C yielded an overall resolution of around 0.13 K. The chip was employed for the screening of the BRCA 2 breast cancer gene; BRCA2 exon 5 was differentiated by its mutant rs80359463 by a 1.1 K difference in melting temperature and two fragments of BRCA2 exon 11 were differentiated by their mutants rs276174826 and rs876660311 by 0.7 K and 2.0 K, respectively. The standard deviations were between 0.1 and 0.5 K. Capable of detecting fluorescence in the DNA and temperature simultaneously and being imaged in a customized assembly, this microchip can be used to screen for mutations in a variety of DNA samples in disease diagnosis and prognosis.

中文翻译：

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一种用于BRCA2突变筛选的DNA熔解曲线快速分析的微流控芯片。

开发了一种集成了微加热器和发光温度传感器的微流控芯片，用于快速，空间融解曲线分析，并将其用于乳腺癌基因片段的筛选。该方法可以在整个过程中总共大约3分钟内检测出遗传差异，这比已建立的过程要快得多。开发了一种微制造技术，以允许将温度感应薄膜和Pt微型加热器与PDMS粘合在一起，并且该芯片可用于通过多光谱光学成像生成和测量热梯度以及染色DNA的荧光强度。传感层由聚（苯乙烯-共-丙烯腈）和三（1,10-菲咯啉）钌（II温度探针是通过在带有附着的Pt微型加热器的玻璃基板上进行刀片涂层而产生的。校准20至90°C的温度可获得约0.13 K的整体分辨率。该芯片用于筛选BRCA 2乳腺癌基因；BRCA2外显子5的突变体rs80359463通过融解温度相差1.1 K来区分，BRCA2外显子11的两个片段分别通过其突变体rs276174826和rs876660311分化了0.7 K和2.0K。标准偏差在0.1到0.5 K之间。该微芯片能够同时检测DNA和温度中的荧光，并在定制的组件中成像，可用于在疾病诊断和预后中筛选各种DNA样品中的突变。