Ultrasensitive microfluidic array for serum pro-inflammatory cytokines and C-reactive protein to assess oral mucositis risk in cancer patients.
Ultrasensitive microfluidic array for serum pro-inflammatory cytokines and C-reactive protein to assess oral mucositis risk in cancer patients.
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血清促炎细胞因子和C反应蛋白的超敏感性微流体阵列评估癌症患者口腔粘膜炎的风险。
DOI：
10.1007/s00216-015-8873-1
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发表时间：
2015-09
影响因子：
4.3
通讯作者：
Rusling JF
中科院分区：
文献类型：
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作者：
Krause CE;Otieno BA;Bishop GW;Phadke G;Choquette L;Lalla RV;Peterson DE;Rusling JF
除了疾病诊断之外，还需要生物标志物来预测癌症治疗副作用如口腔粘膜炎的严重程度。口腔粘膜炎是由高剂量化疗和/或放疗引起的口腔粘膜炎性病变，在口腔癌治疗期间尤其普遍。在这里，我们描述了一个半自动化，模块化的微流控免疫阵列优化的超灵敏度检测的促炎细胞因子参与口腔粘膜炎的病理生物学。我们的目标是在口腔癌治疗的早期，在病变发生之前确定粘膜炎的风险。生物标志物包括肿瘤坏死因子（TNF-α）、白细胞介素-6（IL-6）、白细胞介素-1 β（IL-1β）和C反应蛋白（CRP）。在捕获室中，通过包被有抗体和酶标记的1 μm磁珠从血清中捕获蛋白质分析物。然后将珠子向下游运输到检测室，该检测室含有涂覆有谷胱甘肽-金纳米颗粒（GSH-AuNP）的8传感器阵列和第二组抗体以捕获具有分析物蛋白的珠子。在免疫阵列首次应用于4-蛋白质多重测量中，在5 μL血清中实现了10-40 fg mL−1的超低检测限，可在30分钟内同时检测。质量检测限为2.5-10 zeptomol，低至1500个分子。通过将头颈癌患者血清中4种生物标志物蛋白的水平与标准ELISA的结果相关联，证明了阵列的准确性和诊断实用性。这种方法可能导致快速，低成本的估计癌症患者口腔粘膜炎严重程度的预计风险，以改善治疗管理。
In addition to disease diagnostics, there is a need for biomarkers to predict severity of cancer therapy side effects such as oral mucositis. Oral mucositis is an inflammatory lesion of oral mucosa caused by high dose chemotherapy and/or radiation that is especially prevalent during oral cancer treatment. We describe here a semiautomated, modular microfluidic immunoarray optimized for ultrasensitive detection of pro-inflammatory cytokines involved in pathobiology of oral mucositis. Our goal is methodology to identify risk of mucositis early in oral cancer treatment, before the onset of lesions. Biomarkers include tumor necrosis factor (TNF-α), interleukin-6 (IL-6), interleukin-1β (IL-1β), and C-reactive protein (CRP). Protein analytes were captured from serum in a capture chamber by 1 μm magnetic beads coated with antibodies and enzyme labels. Beads are then transported downstream to a detection chamber containing an 8-sensor array coated with glutathione-gold nanoparticles (GSH-AuNP) and a second set of antibodies to capture the beads with analyte proteins. In this first application of the immunoarray to 4-protein multiplexed measurements, ultralow detection limits of 10–40 fg mL−1 in 5 μL serum were achieved for simultaneous detection in 30 min. Mass detection limits were 2.5–10 zeptomol, as few as 1500 molecules. Accuracy and diagnostic utility of the arrays was demonstrated by correlation of levels of the 4 biomarker proteins in serum from head and neck cancer patients with results from standard ELISA. This approach may lead to rapid, low-cost estimates of projected risk for severity of oral mucositis in cancer patients to enable improved therapeutic management.