Rapid parallel salivary immunoassays on a disposable

描述(申请人提供)：从被采样者的角度来看，采集唾液远比采集血液更可取。然而，近年来，用于测量血液中分析物的微流控技术发展迅速，而唾液作为分析物的使用在被测量的分析物数量和进行这种测量的环境方面都落后了。在一定程度上，这是因为唾液比血浆更易变，分析物的浓度较低，并且含有粘性和粘性粘蛋白。如果将唾液用于通常在血液中测量的许多分析物，并同时对几种分析物进行这些测量是可行的，那么，以一种不需要技术培训的方式，在研究、治疗和健康维护的生物医学测试的质量、频率和范围方面将有可能得到极大的改善，特别是对门诊患者。该项目将开发一种集成的微流控系统，用于快速、廉价和同时测量唾液中的多种分析物，并以简单的一次性聚合物层压形式进行。将开发一种微流控设备，允许从唾液中的粘蛋白中快速提取分析物。两项新的但已得到证实的免疫分析技术将与微流控系统相结合，该系统允许在环境条件下干燥存储所有试剂，并并行测量多个分析物。这些检测方法可以测量低水平的激素、药物、代谢物，甚至是表明疾病存在的蛋白质，以及口腔特有的化合物，如口腔癌的病原体和标志物。免疫分析最初将在已有商业免疫分析的激素上进行验证。正在进行的平行扩散免疫分析的开发工作将通过与粘蛋白提取系统相结合而扩展到唾液检测。为了测量存在的浓度低于扩散免疫分析检测极限的分析物，将使用化学放大的表面等离子体共振成像。利用干试剂的溶解作用，在金表面修饰多个捕获分子。最后，将设计和测试一个多功能的组合系统，允许同时使用这两种方法监测样品。

DESCRIPTION (provided by applicant): The collection of saliva is far preferable to collection of blood from the point of view of the person being sampled. However, in recent years microfluidic technologies for measuring analytes in blood have advanced rapidly, while the use of saliva as an analyte has lagged, both in terms of the number of analytes measured and the environments in which such measurements are made. In part, this is because saliva is more variable than plasma, has analytes in lower concentrations, and contains viscous and adhesive mucins. If it were practical to use saliva for many analytes commonly measured in blood, and to make those measurements on several analytes at once, inexpensively, and in a way not requiring technical training, enormous improvements in the quality, frequency and scope of biomedical testing for research, therapy, and health maintenance would be possible, particularly for ambulatory outpatients. This project will develop an integrated microfluidic system for rapidly, inexpensively, and simultaneously measuring multiple analytes in saliva, and in a simple disposable polymeric laminate format. A microfluidic device to allow rapid extraction of analytes from the mucins in saliva will be developed. Two new but demonstrated immunoassay technologies will be coupled to a microfluidic system that allows dry storage of all reagents at ambient conditions and measures multiple analytes in parallel. These assays can measure low levels of hormones, drugs, metabolites, and even proteins that indicate the presence of disease, as well as compounds specific to the oral cavity such as pathogens and markers for oral cancer. The immunoassays will initially be validated on hormones for which commercial immunoassays are available. Ongoing work on development of parallel diffusion immunoassays will be extended to saliva testing through coupling with the mucin-extraction system. To measure analytes present at concentrations below the limit of detection of the diffusion immunoassay, chemically amplified surface plasmon resonance imaging will be used. Dissolution of dry reagent will be employed to decorate the gold surface with multiple capture molecules. Finally, a versatile combined system will be designed and tested that will allow monitoring of samples by the two methods simultaneously.