Aliquot-level visual indicators of biospecimen exposure to thawed conditions

生物样本暴露于解冻条件下的等分水平视觉指示器

• 批准号: 10560579
• 负责人: CHAD R BORGES
• 金额: $ 17.98万
• 依托单位: ARIZONA STATE UNIVERSITY-TEMPE CAMPUS
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-02-02 至 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10560579
• 关键词: Adherence; Agreement; Aliquot; American; American Society of Clinical Oncology; Archives; Biological; Biomedical Research; Biopsy; Blood; Blood specimen; Chemistry; Clinical; Clinical Research; Collection; Color; Cryopreservation; Custom; Development; Device Designs; Devices; Documentation; ERBB2 gene; Ensure; Estrogen Receptors; Excision; Exposure to; Freezing; Funding; Guidelines; Hour; Ice; Individual; Ischemia; Kinetics; Laboratories; Life; Liquid substance; Magnesium Chloride; Malignant Neoplasms; Oncologist; Operative Surgical Procedures; Oxalates; Paper; Pathologist; Plasma; Process; Quality Control; Reaction; Reagent; Reproducibility; Research Personnel; Running; Sampling; Serum; Specimen; System; Technology; Temperature; Testing; Time; Tissue Fixation; Tissue Inhibitor of Metalloproteinase-1; Tissues; United States National Institutes of Health; Vascular Endothelial Growth Factors; Vial device; Visual; biological specimen archives; cell free DNA; college; cost; design; evidence base; improved; interest; malignant breast neoplasm; notch protein; novel; permanganate; prospective; prototype; quality assurance; sample collection; sample fixation; tool

Project Summary/Abstract Many biological analytes of interest to both clinical oncologists and cancer researchers are unstable when the unfixed biospecimens in which they reside are exposed to thawed conditions. For example, American Society of Clinical Oncology (ASCO)/College of American Pathologists (CAP) guidelines state that the post-excision- up-to-fixation exposure time span for tissues collected for clinical HER2 testing in breast cancer must be less than 1 hour. For blood plasma/serum and many types of tissue specimens that are to be frozen, the proper cold storage temperature is well below the common laboratory freezer temperature of -20 °C. For this and many other reasons, every year improprieties and inconsistencies in pre-analytical sample handling and storage generate unacceptably large numbers of costly false leads in biomedical research. Unsurprisingly, experts in the field agree that this problem must be minimized immediately. Currently there are few tools and not so much as one widely accepted approach by which to implement evidence-based tracking of biospecimen exposure to thawed conditions. In practice, it is actually quite rare for biomedical researchers to employ any evidence-based QA/QC tools at all—which suggests that easy-to-use, individual aliquot-level thawed-state indicators could have a major impact on improving biospecimen quality tracking and therefore actual quality. Here, we will develop color change-based indicators designed for 12 different biospecimen handling/storage requirements that ultimately will be implemented by placement into custom-designed dual-chambered, flex- activated (like a glow-stick) see-through devices that can be housed on (notched into) the outside of a specially designed sample storage vial. Once the bright pink color of permanganate (Mn(VII)) in the indicator turns clear due to its reduction to Mn(II) (as easily seen from the outside of the storage vial), this indicates that the specimen inside the vial has been exposed to a time-temperature span that is outside of the allowed storage/handling parameters. Importantly, subsets of these indicators will not freeze until -33.5 °C or -76 °C, facilitating the unprecedented ability to track exposures to very cold, but nevertheless improper storage temperatures—all in a manner that will be difficult to ignore by anyone handling the samples. Development of these indicators will be accomplished via two specific aims: Specific Aim 1: Establish kinetic control of the permanganate/oxalate reaction system by designing reaction starting conditions so that 12 uniquely useful time/temperature indicating solutions are developed. Specific Aim 2: Verify that reaction kinetics and temperature sensitivities for the 12 indicators behave as expected—that is, A) accurately, precisely (reproducibly), and with substantially increasing run times at 25 °C, 4 °C, 0 °C, and -20 °C—and B) remain accurate and precise when different stock solutions, reagent lots, analysts, or varying periods of freezing are involved.

项目总结/摘要 临床肿瘤学家和癌症研究人员感兴趣的许多生物分析物在肿瘤发生时是不稳定的。 将它们所在的未固定的生物样本暴露于解冻条件。例如，美国社会 临床肿瘤学（ASCO）/美国病理学家学会（CAP）指南指出，切除后- 采集用于乳腺癌临床HER 2检测的组织的固定前暴露时间跨度必须更短 1小时以上。对于待冷冻的血浆/血清和许多类型的组织标本， 冷藏温度远低于-20 °C的普通实验室冷冻温度。为这个和 许多其他原因，每年在分析前样品处理中存在不适当和不一致的情况， 存储在生物医学研究中产生了大量代价高昂错误线索，这是不可接受的。不出意外的是， 该领域的专家一致认为，必须立即尽量减少这一问题。目前，工具很少， 没有一种被广泛接受的方法来实施基于证据的生物样本跟踪 暴露于解冻条件下。在实践中，生物医学研究人员实际上很少雇用任何 基于证据的QA/QC工具-这表明易于使用的单个等分试样解冻状态 指标可对改进生物样本质量跟踪和实际质量产生重大影响。 在这里，我们将开发基于颜色变化的指示剂，用于12种不同的生物标本处理/储存 这些要求最终将通过放置到定制设计的双腔、柔性 激活（如荧光棒），可以容纳在（凹口进入）一个特殊的外部透视设备， 设计的样品储存瓶。一旦指示剂中的高锰酸盐（Mn（VII））呈亮粉色变清晰 由于其还原为Mn（II）（如从储存小瓶的外部容易看到的），这表明 小瓶内的样本已暴露于允许的时间-温度范围之外 储存/处理参数。重要的是，这些指示剂的子集在-33.5 °C或-76 °C之前不会冻结， 促进了前所未有的能力，以跟踪暴露于非常寒冷，但仍然不适当的储存 温度-所有这些都是任何处理样品的人都难以忽视的。 这些指标的制定将通过两个具体目标来完成： 具体目标1：通过设计反应，建立高锰酸盐/草酸盐反应体系的动力学控制 起始条件，从而开发出12种独特有用的时间/温度指示解决方案。 具体目标2：验证12种指示剂的反应动力学和温度敏感性是否表现为 预期的-即，A）准确地，精确地（可再现地），并且在25 °C下运行时间显著增加， 4 °C、0 °C和-20 ° C-和B）在不同储备液、试剂批次 分析师，或不同时期的冻结涉及。