Determination of cerebrospinal fluid leakage by selective deletion of transferrin glycoform using an immunochromatographic assay.,Theranostics

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Determination of cerebrospinal fluid leakage by selective deletion of transferrin glycoform using an immunochromatographic assay.
Theranostics ( IF 12.4 ) Pub Date : 2019-01-01 , DOI: 10.7150/thno.34411
Jusung Oh ₁ , Seok-Joon Kwon _{2,

3} , Jonathan S Dordick _{2,

3} , William J Sonstein ₄ , Robert J Linhardt _{2,

3,

5} , Min-Gon Kim ₁

Affiliation

Cerebrospinal fluid (CSF) leakage can lead to brain and spine pathologies and there is an urgent need for a rapid diagnostic method for determining CSF leakage. Beta-2 transferrin (β2TF), asialotransferrin, is a specific CSF glycoprotein biomarker used to determine CSF leakage when distinguished from serum sialotransferrin (sTF). Methods: We detected β2TF using an immunochromatographic assay (ICA), which can be potentially developed as a point-of-care (POC) testing platform. Sialic acid-specific lectin selectively captures sTF in multiple deletion lines within an ICA test strip, enabling the detection of β2TF. A sample pre-treatment process efficiently captures excess sTF increasing sensitivity for CSF leakage detection. Results: An optimal cut-off value for determining the presence of CSF in test samples was obtained from receiver operating characteristic (ROC) analysis of the ratio of the test signal intensity and the deletion lines. On 47 clinical samples, ICA test strips discriminated CSF positive from negative samples with statistically significant (positive versus negative t-test; P =0.00027). Additional artificial positive samples, prepared by mixing CSF positive and negative clinical samples, were used as a further challenge. These positive samples were clearly discriminated from the negative samples (mixture versus negative t-test; P =0.00103) and CSF leakage was determined with 97.1% specificity and 96.2% sensitivity. Conclusions: ICA represents a promising approach for POC diagnosis of CSF leakage. While requiring 70 min assay time inconvenient for POC testing, our method was significantly shorter than conventional electrophoresis-based detection methods for β2TF.

中文翻译：

通过使用免疫色谱分析法选择性删除转铁蛋白糖型来确定脑脊液渗漏。

脑脊液（CSF）泄漏会导致脑部和脊柱病变，因此迫切需要一种快速的诊断方法来确定CSF泄漏。Beta-2转铁蛋白（β2TF），脱唾液酸转铁蛋白，是一种特殊的CSF糖蛋白生物标志物，用于与血清唾液转铁蛋白（sTF）区别时用于确定CSF渗漏。方法：我们使用免疫色谱分析（ICA）检测到了β2TF，可以将其开发为即时检验（POC）测试平台。唾液酸特异性凝集素选择性地捕获ICA测试条内多个缺失系中的sTF，从而能够检测到β2TF。样品预处理过程可有效捕获过量的sTF，从而提高对CSF泄漏检测的敏感性。结果：从测试信号强度与缺失线之比的接收器工作特性（ROC）分析中获得了用于确定测试样品中CSF的存在的最佳临界值。在47个临床样品上，ICA试纸将CSF阳性与阴性样品区分开，具有统计学显着性（阳性与阴性t检验； P = 0.00027）。通过混合CSF阳性和阴性临床样品制备的其他人工阳性样品被用作进一步的挑战。这些阳性样品与阴性样品有明显区别（混合物与阴性t检验； P = 0.00103），CSF漏出的特异性为97.1％，灵敏度为96.2％。结论：ICA代表了一种有前途的POC诊断脑脊液漏的方法。尽管需要70分钟的分析时间来进行POC测试很不方便，

更新日期：2019-01-01

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