Furuya Hideki, Sakatani Toru, Tanaka Sunao, Murakami Kaoru, Waldron Richard T, Hogrefe Wayne, Rosser Charles J
Cedars-Sinai Medical Center.
Cedars-Sinai Comprehensive Cancer Center: Cedars-Sinai Medical Center Samuel Oschin Comprehensive Cancer Institute.
Res Sq. 2023 Nov 25:rs.3.rs-3635581. doi: 10.21203/rs.3.rs-3635581/v1.
No single marker of bladder cancer (BC) exists in urine samples with sufficient accuracy for disease diagnosis and treatment monitoring. The multiplex Oncuria BC assay noninvasively quantifies the concentration of 10 protein analytes in voided urine samples to quickly generate a unique molecular profile with proven BC diagnostic and treatment-tracking utility. Test adoption by diagnostic and research laboratories mandates reliably reproducible assay performance across a variety of instrumentation platforms used in different laboratories.
We compared the performance of the clinically validated Oncuria BC multiplex immunoassay when data output was generated on three different analyzer systems. Voided urine samples from 36 subjects (18 with BC and 18 Controls) were reacted with Oncuria test reagents in three 96-well microtiter plates on Day 1, and consecutively evaluated on the LED/image-based MagPix, and laser/flow based Luminex 200 and FlexMap 3D (all xMAP instruments from Luminex Corp., Austin, TX) on Day 2. The BC assay uses magnetic bead-based fluorescence technology (xMAP, Multi-analyte profiling; Luminex) to simultaneously quantify 10 protein analytes in urine specimens [i.e., angiogenin (ANG), apolipoprotein E (ApoE), carbonic anhydrase IX (CA9), CXCL8/interleukin-8 (IL-8), matrix metalloproteinase-9 (MMP-9), matrix metalloproteinase-10 (MMP-10), serpin A1/alpha-1 anti-trypsin (A1AT), serpin E1/plasminogen activator inhibitor-1 (PAI-1), CD138/syndecan-1 (SDC1), and vascular endothelial growth factor-A (VEGF-A)].
All three platforms categorized all 10 analytes in identical samples at nearly identical concentrations, with variance across systems typically <5%. While the most contemporary instrument, the FlexMap 3D, output higher raw fluorescence values than the two comparator systems, standard curve slopes and analyte concentrations determined in urine samples were concordant across all three units. Forty-four percent of BC samples registered ≥1 analyte above the highest standard concentration, i.e., A1AT (n=7/18), IL-8 (n=5), and/or ANG (n=2). In Controls, A1AT was higher in one sample.
Multiplex BC assays generate detailed molecular signatures useful for identifying BC, predicting treatment esponsiveness, and tracking disease progression and recurrence. The similar performance of the Oncuria assay across three different analyzer systems supports test adaptation by clinical and research laboratories using existing xMAP platforms.
This study was registered at ClinicalTrials.gov as NCT04564781, NCT03193528, NCT03193541, and NCT03193515.
在尿液样本中，不存在一种对膀胱癌（BC）诊断和治疗监测具有足够准确性的单一标志物。Oncuria BC多重检测可对晨尿样本中的10种蛋白质分析物进行无创定量，以快速生成具有已证实的BC诊断和治疗跟踪效用的独特分子图谱。诊断和研究实验室采用该检测方法要求在不同实验室使用的各种仪器平台上具有可靠的可重复检测性能。
我们比较了在三种不同分析仪系统上生成数据输出时，经过临床验证的Oncuria BC多重免疫检测的性能。来自36名受试者（18名BC患者和18名对照）的晨尿样本在第1天与Oncuria检测试剂在三个96孔微量滴定板中反应，并在第2天在基于LED/图像的MagPix以及基于激光/流式的Luminex 200和FlexMap 3D（所有xMAP仪器均来自德克萨斯州奥斯汀的Luminex公司）上进行连续评估。BC检测使用基于磁珠的荧光技术（xMAP，多分析物谱分析；Luminex）同时定量尿液样本中的10种蛋白质分析物[即血管生成素（ANG）、载脂蛋白E（ApoE）、碳酸酐酶IX（CA9）、CXCL8/白细胞介素-8（IL-8）、基质金属蛋白酶-9（MMP-9）、基质金属蛋白酶-10（MMP-10）、丝氨酸蛋白酶抑制剂A1/α-1抗胰蛋白酶（A1AT）、丝氨酸蛋白酶抑制剂E1/纤溶酶原激活物抑制剂-1（PAI-1）、CD138/多配体蛋白聚糖-1（SDC1）和血管内皮生长因子-A（VEGF-A）]。
所有三个平台对相同样本中的所有10种分析物进行分类时，浓度几乎相同，各系统之间的差异通常<5%。虽然最新型的仪器FlexMap 3D输出的原始荧光值高于两个比较系统，但在所有三个仪器上，尿液样本中测定的标准曲线斜率和分析物浓度是一致的。44%的BC样本记录到≥1种分析物高于最高标准浓度，即A1AT（n = 7/18）、IL-8（n = 5）和/或ANG（n = 2）。在对照中，有一个样本的A1AT较高。
多重BC检测可生成有助于识别BC、预测治疗反应以及跟踪疾病进展和复发的详细分子特征。Oncuria检测在三种不同分析仪系统上的相似性能支持使用现有xMAP平台的临床和研究实验室采用该检测方法。
本研究已在ClinicalTrials.gov注册，注册号为NCT04564781、NCT03193528、NCT03193541和NCT03193515。
