Core D: Molecular Diagnostics

核心、目标I/II、III涉及来自 单个多中心研究中心。到目前为止，我们已经收到了1,143名参加试验的患者， 符合RT-PCR随机化条件，其中29例不合格。其中274例为PCR+（26%）， PCR阴性（74%）。40例患者不合格，21例仍在等待结果。PE组织切片 按照SOP中的记录制备的样品从美国或世界各地的不同地点发送至 核心，并进行PCR分析。这些研究已按计划进行。所有 各中心及时收到结果。总的来说，研究中没有出现大的问题 工作流已与每个中心进行讨论，以改善组织的处理和运输 标本对于未遵守我们的指南或样本质量较差的中心， 从PCR随机化研究中退出。 对于目标IV，我们正在从JWCI和MLST-I试验中收集PE和/或IHC阳性SLN组织块 中心.将组织块切片用于免疫组织化学（IHC）和RNA分离。各节 正在项目II中用于评估新的黑素瘤IHC和RT-PCR生物标志物。 对于目标V，我们一直在分离和定量来自SLN、非SLN和转移性黑色素瘤的RNA 组织中分离RNA并冷冻保存用于项目II。超过200个组织样本 已处理。这些标本已成为开发和验证新的 mRNA标记物。此外，我们还收到了250多个原发性和转移性肿瘤标本， 悉尼黑色素瘤单位进行RNA分析。这些石块被送到我们这里，切成段，加工， 送回给他们。 这些标本对于我们没有特定类型的黑色素瘤或 转移到特定部位。 第六，我们继续改善标准作业程序及精简运作。我们扩大了 我们所有样本库的ItemTracker程序。Microsoft SQL Server用作数据库 管理引擎，它使ItemTracker能够输入和存储数百万条记录。系统直观 视觉驱动的用户界面结合了条形码、标签制作、数据过滤、数据字段 自定义、存储布局的可视化表示等等。所有标本在 冻死了我们在每个冷冻室都有一个虚拟的标本跟踪器来监控储存情况。 在Aim VII中，我们累积了121例原发性黑色素瘤。中心提供了组织块， 核心切割组织进行IHC和DNA/RNA分析。块返回到中心 月正在收集黑色素瘤的特定亚型。这些标本正在项目II中使用。 对样品进行了DMA分析和IHC，结果显示质量良好。研究 如CCR 9受体mRNA分析。这是最近出版的 发表在《临床癌症研究》上，并接受了特别的编辑评论。原发性肿瘤组织也用于 甲基化肿瘤相关DMA标志物的评估。核心使用 配备紫外线和激光的Veritas Arcturus系统。 对于目的VII，从进入MSLT-II试验的患者中采集血清。血清有 进行分子/遗传分析。血液被收集用于分子研究， MLST站点。接收血液并在特定的分子SOP下处理，然后冷冻储存。我们 有1,836例多中心出血。血液中的血清经过处理，将用于DMA 研究项目二。 目标八将不再执行，因为这一需要不再是主要优先事项， 可用资金。

The Core, Aims I/I I, III are involved in the accrual of patients with paraffin-embedded (PE) SLNs from the individual multicenter sites. To date, we have received 1,143 patients who have entered into the trial that were qualified for RT-PCR randomization of which 29 were ineligible. Of these 274 were PCR+ (26%) and 794 were PCR- (74%). Forty patients were ineligible and 21 are still pending results. The PE tissue sections prepared as documented in the SOPs were sent from different sites in the USA or around the world to the core and been processed for PCR analysis. The studies have been running accordingly as planned. All centers are receiving results in a timely manner. Overall, there have been no major problems in the study workflow. Discussions have been carried out with each center to improve processing and shipping of tissue specimens. For centers which have not complied with our guidelines nor have poor quality specimens, were dropped from the PCR randomized study. For Aim IV, we are collecting PE and/or IHC positive SLN blocks from JWCI and from the MLST-I trial centers. The blocks have been sectioned for immunohistochemistry (IHC) and RNA isolation. The sections are being used to assess new melanoma IHC and RT-PCR biomarkers in Project II. For Aim V, we have been isolating and quantifying RNA from SLN, non-SLN and metastatic melanoma tissues. The RNA is isolated and cryopreserved for Project II. Over 200 tissue specimens have been processed. These specimens have served as a valuable resource for development and validation of new mRNA markers. In addition, we have received over 250 primary and metastatic tumor specimens from the Sydney Melanoma Unit for RNA analysis. The blocks have been sent to us, cut into sections, processed and sent back to them. These specimens have been important for studies we do not have particular types of melanomas or metastasis to specific sites. Aim VI, we have continued to improve the SOPs and streamline operations. We have expanded the ItemTracker program for all our specimen banking. Microsoft SQL Server serves as the database management engine, which enables ItemTracker to input and store millions of records. The system's intuitive visually-driven user interface incorporates functionality for barcodes, label making, data filtering, data fields customization, visual representation of storage layouts, and much more. All specimens are barcoded prior to freezing. We have a virtual specimen tracker in each freezer to monitor storage. In Aim VII, we have accrued 121 primary melanomas. Centers have provided tissue blocks whereby the Core cuts the tissues for IHC, and DNA/RNA analysis. The blocks are returned back to the center within a month. Specific subtypes of melanoma are being collected. These specimens are being used in Project II. DMA analysis and IHC have been performed on the samples and have shown good quality results. Studies such as the CCR9 receptor mRNA analysis was performed on these specimens. This was recently published in Clin Cancer Res and received a special editorial review. The primary tumor tissue was also used for assessment of methylated tumor-related DMA markers. The core performs laser microdissection using the Veritas Arcturus system equipped with both UV and laser light. For Aim VII, serum has been collected from the patients entered into the MSLT-II trial. The serum has been processed for molecular/genetic analysis. Bloods are being collected for molecular studies from selected MLST sites. The blood is received and processed under a specific molecular SOP and then cryostored. We have 1,836 bleeds from the multicenter sites. The bloods' serum is processed and will be used for DMA studies in Project II. Aim VIII will not be performed any longer as the need for this is not a major priority any longer due to funds available.