Faculty of Arts and Science, McMaster University, Hamilton, ON, Canada.
Niagara Health Knowledge Institute, St. Catharines, ON, Canada.
PLoS One. 2024 Oct 31;19(10):e0311921. doi: 10.1371/journal.pone.0311921. eCollection 2024.
Biosampling studies in critically ill patients traditionally involve bedside collection of samples followed by local processing (ie. centrifugation, aliquotting, and freezing) and storage. However, community hospitals, which care for the majority of Canadian patients, often lack the infrastructure for local processing and storage of specimens. A potential solution is a "simplified" biosampling protocol whereby blood samples are collected at the bedside and then shipped to a central site for processing and storage. One potential limitation of this approach is that delayed processing may alter sample characteristics.
To determine whether delays in blood sample processing affect the stability of cytokines (IL-6, TNF, IL-10, IFN-γ), chemokines (IL-8, IP-10, MCP-1, MCP-4, MIP-1α, MIP-1β), cell-free DNA (cfDNA) (released by dying cells), and blood clotting potential in human blood samples.
Venous blood was collected into EDTA and citrate sample tubes and stored at room temperature (RT) or 4°C for progressive intervals up to 72 hours, prior to processing. Plasma cytokines and chemokines were quantified using single or multiplex immunoassays. cfDNA was measured using Picogreen DNA Quantification. Blood clotting potential was measured using a thrombin generation assay.
Blood samples were collected from 9 intensive care unit (ICU) patients and 7 healthy volunteers. Admission diagnoses for the ICU patients included sepsis, trauma, ruptured abdominal aortic aneurysm, intracranial hemorrhage, gastrointestinal bleed, and hyperkalemia. After pre-processing delays of up to 72 hours at RT or 4°C, no significant changes were observed in plasma cytokines, chemokines, cfDNA, or thrombin formation.
Delayed sample processing for up to 72 hours at either RT or 4°C did not significantly affect cytokines, chemokines, cfDNA, or blood clotting potential in plasma samples from healthy volunteers and ICU patients. A "simplified" biosampling protocol is a feasible solution for conducting biosampling research at hospitals without local processing capacity.
传统上，对危重症患者进行生物采样研究涉及在床边采集样本，然后进行局部处理（即离心、分装和冷冻）和储存。然而，为大多数加拿大患者提供护理的社区医院通常缺乏对标本进行局部处理和储存的基础设施。一种潜在的解决方案是“简化”生物采样方案，即在床边采集血液样本，然后运送到中心站点进行处理和储存。这种方法的一个潜在限制是，延迟处理可能会改变样本特征。
确定血液样本处理的延迟是否会影响细胞因子（IL-6、TNF、IL-10、IFN-γ）、趋化因子（IL-8、IP-10、MCP-1、MCP-4、MIP-1α、MIP-1β）、无细胞 DNA（cfDNA）（由死亡细胞释放）和人血样中凝血潜力的稳定性。
将静脉血采集到 EDTA 和柠檬酸盐采样管中，并在室温（RT）或 4°C 下储存，直至处理前达到 72 小时的不同时间点。使用单或多指标免疫测定法定量检测血浆细胞因子和趋化因子。使用 Picogreen DNA 定量法测量 cfDNA。使用凝血酶生成测定法测量凝血潜力。
从 9 名重症监护病房（ICU）患者和 7 名健康志愿者中采集了血液样本。ICU 患者的入院诊断包括败血症、创伤、腹主动脉瘤破裂、颅内出血、胃肠道出血和高钾血症。在 RT 或 4°C 下预处理延迟长达 72 小时后，未观察到血浆细胞因子、趋化因子、cfDNA 或凝血酶形成有显著变化。
在 RT 或 4°C 下延迟长达 72 小时的样本处理不会显著影响健康志愿者和 ICU 患者血浆样本中的细胞因子、趋化因子、cfDNA 或血液凝固潜力。“简化”的生物采样方案是在没有本地处理能力的医院进行生物采样研究的可行解决方案。
