Department of Neurology, University Hospital Ostrava, Ostrava, Czech Republic; Department of Clinical Neurosciences, University of Ostrava, Ostrava, Czech Republic.
Institute of Laboratory Medicine, University Hospital Brno, Brno, Czech Republic.
Mult Scler Relat Disord. 2022 Nov;67:104177. doi: 10.1016/j.msard.2022.104177. Epub 2022 Sep 11.
Serum neurofilaments (sNfs), especially the most investigated serum neurofilament light chain (sNfL), are promising biomarkers in multiple sclerosis (MS). However, their clinical utility is still limited, given the availability and costs of accessible analytical methods. The gold standard for the detection of sNfs is represented by the single molecule arrays (SIMOA). Recently, a high sensitivity enzyme-linked immunosorbent assay (hsELISA) has also been introduced. The objective of the study was to compare both assays for the determination of sNfL and neurofilament heavy chain (sNfH) concentrations in a defined MS cohort. The second objective was to identify contributing factors to sNfs concentrations determined by hsELISA.
Serum samples were collected from MS patients attending the MS Centre, University Hospital Ostrava, Czech Republic. The levels of sNfs were detected using SIMOA and hsELISA assays.
The Spearman's rank correlation coefficient between the sNfL SIMOA and sNfL hsELISA and between the sNfH SIMOA and sNfH hsELISA was moderate r= 0.543 (p = 0.001) and r= 0.583 (p = 0.001), respectively. The Passing-Bablok regression analysis demonstrated bias between both methods. Equally significant bias between the methods was confirmed by the Bland-Altman plots. Furthermore, confounding factors affecting the sNfL levels were glomerular filtration rate (eGFR; 95% CI -2.34 to -0.04) and sex (95% CI -2.38 to -0.10). The sNfH levels were affected by age (95% CI 0.01 to 0.07), eGFR (95% CI -2.45 to -0.02), body mass index (BMI; 95% CI -0.31 to -0.05), and blood volume (95% CI 0.69 to 3.35).
This analytical study showed significant differences between hsELISA and SIMOA methods, especially for the sNfH concentrations. We identified confounding factors for sNfs levels determined by hsELISA. The sNfs levels were influenced by renal function and sex, whilst sNfH levels were affected by age, BMI, and total blood volume.
血清神经丝（sNfs），尤其是研究最多的血清神经丝轻链（sNfL），是多发性硬化症（MS）有前途的生物标志物。然而，由于可及的分析方法的可用性和成本，其临床实用性仍然有限。sNfs 的金标准检测方法是单分子阵列（SIMOA）。最近，也引入了一种高灵敏度酶联免疫吸附测定法（hsELISA）。本研究的目的是比较这两种测定 sNfL 和神经丝重链（sNfH）浓度的方法在一个明确的 MS 队列中的应用。第二个目的是确定 hsELISA 测定的 sNfs 浓度的影响因素。
从捷克奥斯特拉瓦大学医院 MS 中心就诊的 MS 患者中采集血清样本。使用 SIMOA 和 hsELISA 测定法检测 sNfs 水平。
sNfL SIMOA 和 sNfL hsELISA 以及 sNfH SIMOA 和 sNfH hsELISA 之间的 Spearman 秩相关系数为中度 r=0.543（p=0.001）和 r=0.583（p=0.001）。通过 Passing-Bablok 回归分析证实了两种方法之间存在偏差。Bland-Altman 图也证实了两种方法之间存在显著偏差。此外，影响 sNfL 水平的混杂因素是肾小球滤过率（eGFR；95%CI-2.34 至-0.04）和性别（95%CI-2.38 至-0.10）。sNfH 水平受年龄（95%CI0.01 至 0.07）、eGFR（95%CI-2.45 至-0.02）、体重指数（BMI；95%CI-0.31 至-0.05）和血容量（95%CI0.69 至 3.35）的影响。
这项分析研究表明 hsELISA 和 SIMOA 方法之间存在显著差异，特别是对于 sNfH 浓度。我们确定了 hsELISA 测定的 sNfs 水平的混杂因素。sNfs 水平受肾功能和性别影响，而 sNfH 水平受年龄、BMI 和总血容量影响。
