基于表面增强拉曼光谱法的微流控芯片检测肿瘤外泌体用于卵巢癌精准诊断

Cancer cell-derived exosomes carry a large amount of molecular information of their maternal tumor. They are also involved in the regulation of tumor development and metastasis, and thus, are regarded as a new type of biomarker for early cancer diagnosis. Ovarian cancer is a serious threat to women's life and health, so it is very important to achieve early diagnosis. However, there is still no effective exosome detection method for accurate clinical diagnosis of ovarian cancer. This project is aimed to develop a technology, based on the combination of microfluidic chip and Surface-enhanced Raman Spectroscopy (SERS), for the detection of tumor exosomes in body fluids, which will lead to the realization of accurate early diagnosis of ovarian cancer. Due to the advantages of the microfluidic chip including the small size, small volume of samples required, and the precise control of liquid, exosomes can be immunocaptured by modifying the specific antibodies on the gold substrate in the microfluidic channels. Afterwards, by using the SERS probes which conjugated with Raman reporter molecules and exosome membrane antibodies, the captured exosomes can be recognized to achieve highly sensitive detection. The designed microfluidic chip has a parallel multi-channel structure, which can simultaneously achieve specific, parallel and independent detection of various membrane protein markers, thereby achieving clinically required high-throughput detection and improving the accuracy of early ovarian cancer diagnosis. The results of this study are expected to establish a new biopsy method based on patient's body fluids, which provides research basis for clinical early accurate diagnosis of ovarian cancer, as well as the effective monitoring of postoperative cancer metastasis.

癌细胞外泌体携带大量母体肿瘤分子信息，是一种癌症早期诊断新型标志物，其参与调控肿瘤的发展和转移。卵巢癌严重威胁女性的生命与健康，实现其早期诊断至关重要，但目前临床仍缺乏有效的外泌体检测手段用于卵巢癌的精准诊断。本项目拟基于表面增强拉曼光谱法（SERS）开发一种微流控芯片检测技术用于检测体液中的肿瘤外泌体，以实现卵巢癌早期精准诊断。借助微流控芯片尺寸小、所需样品少、对液体精确控制的优势，通过在微流通道内的金基底上修饰特异性抗体后免疫捕获外泌体，然后利用缀合拉曼报告分子和膜蛋白抗体的SERS探针识别捕获外泌体以实现高灵敏检测。所设计的微流控芯片具有平行多通道结构，可以同时实现多种膜蛋白标志物的特异性、平行、独立检测，从而达到临床要求的高通量检测并提高早期卵巢癌诊断结果的准确性。研究结果有望建立基于患者体液的活检新方法，期冀为临床卵巢癌的早期精准诊断和术后癌细胞转移的有效监测提供研究基础。

外泌体是一种细胞分泌的纳米级脂质膜囊泡，其参与细胞间信息交流和信号传导。癌细胞外泌体携带大量母体肿瘤分子信息，参与调控肿瘤的生长发展和侵袭转移等过程。卵巢恶性肿瘤严重威胁女性的生命与健康，因此，开发基于体液中外泌体检测的无创即时诊断，并通过肿瘤细胞分泌的外泌体获取肿瘤细胞发生发展的信息非常重要。本项目开发了一种便携式微流控芯片设备，借助于先进的光谱检测技术如表面增强拉曼光谱法（SERS）进行体液中肿瘤外泌体的检测，以实现肿瘤如卵巢癌的早期精准诊断。微流控芯片具有经济、便携、对小体积液相样品精确控制的特点而成为外泌体检测的理想平台，而SERS又具有灵敏度高、非侵入性的优势。结合微流控技术和SERS 标签的检测方法可以解决目前临床卵巢癌早期诊断技术缺乏的难题。借助微流控芯片尺寸小、所需样品少、对液体精确控制的优势，通过在微流通道内的金基底上修饰特异性抗体后免疫捕获外泌体，然后利用缀合拉曼报告分子和膜蛋白抗体的SERS探针识别捕获外泌体实现高灵敏检测。所设计的微流控芯片具有平行多通道结构，可以同时实现多种膜蛋白标志物的特异性、平行、独立检测，从而达到临床要求的高通量检测并提高早期卵巢癌诊断结果的准确性。研究结果开发了基于患者体液的活检新方法，有望为临床卵巢癌的早期精准诊断提供研究基础。该项技术的成功发展将加速外泌体的研究，为开发新的卵巢癌液体活检技术和临床癌症早期精准诊断提供可靠的研究基础。