A Sensitive and Automated Diagnostic Assay for Lung Cancer

描述（由申请人提供）：肺癌的灵敏和自动诊断测定肺癌是美国的头号癌症杀手。在早期可治愈阶段识别肺癌的能力将降低死亡率，因此是一个重要的临床目标。为了实现这一目标，我们已经确定了一组遗传生物标志物，可以在痰液中检测到肺癌的早期检测（AACR突发新闻）。我们还开发了一种原位微型芯片，包括多个荧光素标记的探针，可以在一次测试中测量遗传生物标志物。然而，探针的荧光信号很弱，衰减快，降低了生物标志物的灵敏度。此外，手动计数多个探头的模糊信号是耗时且劳动密集的，具有高可变性，因此限制了临床环境中有前途的诊断方法。最近的研究表明，荧光团-金属相互作用，即，金属增强荧光（MEF），可以增加荧光信号的强度和光稳定性，并且新兴的自动点计数系统允许快速和客观地分析多个探针。R21探索性项目的目标是开发原位微型芯片作为一种可靠的诊断方法，可以可靠地检测痰液样本中肺癌早期检测的遗传生物标志物。我们建议1）通过将荧光探针与固定在涂有银纳米颗粒的载玻片上的细胞杂交来增加原位微型芯片的灵敏度，2）通过使用自动点计数系统分析痰细胞中增强的荧光信号来开发原位微型芯片作为稳健的测定，和3）在我们现有的I期肺癌患者病例对照队列中使用痰样品确定用于肺癌早期检测的稳健测定的诊断性能。完成后，原位微型芯片将被完善为肺癌早期检测的可靠，自动化和非侵入性基因检测。本研究将为大型前瞻性临床项目（R 01）或多中心合作临床研究（R33）奠定基础，旨在进一步评估其全面效用，使其能够在常规临床实验室中快速广泛接受和应用。这项探索性研究将汇集在分子遗传学，纳米技术，细胞病理学，肿瘤学和生物统计学方面具有广泛专业知识的科学家和临床医生的互补优势。

DESCRIPTION (provided by applicant): A Sensitive and Automated Diagnostic Assay for Lung Cancer Lung cancer is the number one cancer killer in the USA. The ability to identify lung cancer at its early, curable stages will reduce the mortality and is thus an important clinical goal. Toward this goal, we have identified a panel of genetic biomarkers that can be detected in sputum for early detection of lung cancer (AACR Breaking News). We have also developed an in situ mini-chip comprising multiple fluorescein-labeled probes that can measure the genetic biomarkers in a single test. However, the fluorescent signals of the probes are weak and fade fast, reducing the sensitivity of the biomarkers. Furthermore, manually counting the blurry signals of the multiple probes is time- consuming and labor intensive with high variability, thus limiting the promising diagnostic approach in clinical setting. The recent studies have shown that fluorophore-metal interaction, namely, metal-enhanced fluorescence (MEF), can increase intensity and photostability of fluorescent signals, and an emerging automated dot counting system permits rapidly and objectively analyze multiple probes. The objective of the R21 exploratory project is to develop the in situ mini-chip as a robust diagnostic assay that can reliably detect the genetic biomarkers for the early detection of lung cancer in sputum samples. We propose to 1) increase the sensitivity of the in situ mini-chip by hybridizing the fluorescent probes to the cells immobilized on the slides coated with silver nanoparticles, 2) develop the in situ mini-chip as robust assay by analyzing the enhanced fluorescent signals in sputum cells using the automated dot-counting system, and 3) determine diagnostic performance of the robust assay for early detection of lung cancer using sputum samples in our existing case-control cohort of patients with stage I lung cancer. At completion, the in situ mini-chip will be refined as a reliable, automated, and noninvasive genetic test for lung cancer early detection. The study will lay the basis for large prospective clinical project (R01) or multicenter cooperative clinical study (R33) designed to further evaluate its full utility that could rapidly and widely be accepted and applied in routine clinical laboratories. This exploratory study will bring together complementary strengths of scientists and clinician with extensive expertise in molecular genetics, nanotechnology, cytopathology, oncology, and biostatistics.