当前位置: X-MOL 学术Clin. Chem. › 论文详情
Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)
B-316 Evaluation of an FDA-cleared Chromogranin A Assay on an Automated Analyzer
Clinical Chemistry ( IF 7.1 ) Pub Date : 2024-10-02 , DOI: 10.1093/clinchem/hvae106.673
N Boyert, G Schroeder, M Dee, J M Colon-Franco

Background Chromogranin A (CgA) is a hydrophilic and acidic protein, present in the chromaffin granules of neuroendocrine cells. It is the preferred tumor marker for the monitoring of neuroendocrine tumors (NETS), which are often found in the lungs, appendix, pancreas, and gastrointestinal tract. Previous CgA assays were performed by manual ELISA testing, and were not FDA-approved, leading to a longer, more rigorous validation process and longer run times. The introduction of an automated, FDA-cleared CgA assay allows the laboratory to process patient samples more quickly and reliably, shortening a multiple hour process to minutes. The objective of this study was to evaluate the performance of an automated CgA assay recently cleared by the FDA. Methods We validated the the B·R·A·H·M·S™ Chromogranin A II (CGAII) assay on the automated B·R·A·H·M·S KRYPTOR Compact PLUS (Thermo Fisher Scientific Inc.). The CGAII assay is an immunofluorescent assay, which uses Time-Resolved Amplified Cryptate Emission (TRACE) to quantitate CGA in human serum. Assay performance characteristics were evaluated including measuring range (MR) and maximum dilution, precision, method comparison, stability, reference interval, stability, and carryover. Results The MR for CGAII was established with triplicate analysis of serum at 8 levels, ranging from 25.8 - 2,700 ng/mL. Linearity was demonstrated with a slope of 0.907, intercept of 1.948 and observed error of 2.54 ng/mL. Recovery at each point ranged from 83.3-110.3%. Carryover (<5.0 ng/mL) was not observed at a concentration approximately 2 times the upper limit of the MR. A 1:25 or 1:500 automatic dilution was determined to be acceptable (% bias <16%). Intraday precision coefficient of variation (CV) ranged from 4.1-1.7% (SDs from 2.32-6.64 ng/mL) at concentrations of 56.7 and 393.0 ng/mL. The interday CVs ranged from 6.0-7.1% (SDs from 3.43 - 28.84 ng/mL) at concentrations of 57.5 and 403.8 ng/mL. Twenty samples with concentrations ranging from 56.4-2,907 ng/mL were compared to a reference laboratory utilizing a similar methodology. The correlation coefficient was 0.9934, with a slope of 1.014, and intercept of -32.95. An additional twenty samples with concentrations ranging from 67.0-2,446 ng/mL were compared to a CgA ELISA kit (CGA-ELISA-NG, CISBIO) to determine the bias between the two methodologies. The correlation coefficient was 0.9902, with a slope of 0.845, and intercept of -15.31, and a bias of -19.9%. The serum samples were stable for 3 days when stored at 2-8°C. Recovered CgA concentrations fell outside of the 95% confidence interval after day 3 at 2-8°C. The manufacturer provided reference interval of <187 ng/mL was verified by testing 20 healthy adult samples, all of which fell within this range. Conclusions This assay has been validated and shown to be an accurate and precise method for monitoring CgA. Further, the method was validated on an automated analyzer to allow expeditious turnaround time in a core laboratory.

中文翻译:


B-316 在自动分析仪上评估 FDA 批准的嗜铬粒蛋白 A 测定



背景嗜铬粒蛋白 A (CgA) 是一种亲水性酸性蛋白,存在于神经内分泌细胞的嗜铬颗粒中。它是监测神经内分泌肿瘤 (NETS) 的首选肿瘤标志物,神经内分泌肿瘤 (NETS) 常见于肺部、阑尾、胰腺和胃肠道。以前的 CgA 分析是通过手动 ELISA 检测进行的,并且未获得 FDA 批准,导致验证过程更长、更严格,运行时间更长。引入经 FDA 批准的自动化 CgA 检测使实验室能够更快、更可靠地处理患者样本,将数小时的流程缩短到几分钟。本研究的目的是评估 FDA 最近批准的自动化 CgA 检测的性能。方法 验证 B·R·A·H·™ 自动化B·R·A·H·M·S KRYPTOR Compact PLUS (Thermo Fisher Scientific Inc.).CGAII 检测是一种免疫荧光检测,它使用时间分辨扩增隐窝发射 (TRACE) 来定量人血清中的 CGA。评估分析性能特征,包括测量范围 (MR) 和最大稀释度、精密度、方法比较、稳定性、参比区间、稳定性和残留。结果 CGAII 的 MR 是通过对 8 个水平的血清进行三次重复分析建立的,范围为 25.8 - 2,700 ng/mL。线性表现为 0.907,截距为 1.948,观察到的误差为 2.54 ng/mL。每个点的恢复率在 83.3-110.3% 之间。在浓度约为 MR 上限的 2 倍时未观察到残留 (<5.0 ng/mL)。确定 1:25 或 1:500 自动稀释度是可接受的(% 偏差 <16%)。日内精密变异系数 (CV) 范围为 4.1-1.7%(SD 为 2.32-6。64 ng/mL),浓度为 56.7 和 393.0 ng/mL。浓度为 57.5 和 403.8 ng/mL 的日间 CV 范围为 6.0-7.1% (SD 为 3.43 - 28.84 ng/mL)。将浓度范围为 56.4-2,907 ng/mL 的 20 份样品与采用类似方法的参比实验室进行了比较。相关系数为 0.9934,斜率为 1.014,截距为 -32.95。将另外 20 个浓度范围为 67.0-2,446 ng/mL 的样品与 CgA ELISA 试剂盒(CGA-ELISA-NG、CISBIO)进行比较,以确定两种方法之间的偏差。相关系数为 0.9902,斜率为 0.845,截距为 -15.31,偏差为 -19.9%。血清样品在 2-8°C 下储存 3 天。 在 2-8°C 下第 3 天后,回收的 CgA 浓度超出 95% 置信区间。 制造商提供的参考区间为 <187 ng/mL,通过测试 20 个健康成人样品进行验证,所有这些样品均在此范围内。结论 该测定法已经过验证,并被证明是一种准确且精密的 CgA 监测方法。此外,该方法在自动分析仪上进行了验证,可在核心实验室中实现快速周转时间。
更新日期:2024-10-02
down
wechat
bug