Zhang Xiao Guang, Shu Yao Gen, Gao Ju, Wang Xuan, Liu Li Peng, Wang Meng, Cao Yu Xi, Zeng Yi
National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China.
Beijing Runbio Biotechnology Development Co., Ltd, Beijing 100106, China.
Biomed Environ Sci. 2017 Jun;30(6):426-431. doi: 10.3967/bes2017.056.
To develop a rapid, highly sensitive, and quantitative method for the detection of NT-proBNP levels based on a near-infrared point-of-care diagnostic (POCT) device with wide scope.
The lateral flow assay (LFA) strip of NT-proBNP was first prepared to achieve rapid detection. Then, the antibody pairs for NT-proBNP were screened and labeled with the near-infrared fluorescent dye Dylight-800. The capture antibody was fixed on a nitrocellulose membrane by a scribing device. Serial dilutions of serum samples were prepared using NT-proBNP-free serum series. The prepared test strips, combined with a near-infrared POCT device, were validated by known concentrations of clinical samples. The POCT device gave the output of the ratio of the intensity of the fluorescence signal of the detection line to that of the quality control line. The relationship between the ratio value and the concentration of the specimen was plotted as a work curve. The results of 62 clinical specimens obtained from our method were compared in parallel with those obtained from the Roche E411 kit.
Based on the log-log plot, the new method demonstrated that there was a good linear relationship between the ratio value and NT-proBNP concentrations ranging from 20 pg/mL to 10 ng/mL. The results of the 62 clinical specimens measured by our method showed a good linear correlation with those measured by the Roche E411 kit.
The new LFA detection method of NT-proBNP levels based on the near-infrared POCT device was rapid and highly sensitive with wide scope and was thus suitable for rapid and early clinical diagnosis of cardiac impairment.
基于具有广泛应用范围的近红外即时诊断(POCT)设备,开发一种快速、高灵敏度且定量检测NT - proBNP水平的方法。
首先制备NT - proBNP的侧向流动分析(LFA)试纸条以实现快速检测。然后,筛选NT - proBNP的抗体对并用近红外荧光染料DyLight - 800进行标记。捕获抗体通过划线装置固定在硝酸纤维素膜上。使用不含NT - proBNP的血清系列制备血清样本的系列稀释液。将制备好的试纸条与近红外POCT设备结合,用已知浓度的临床样本进行验证。POCT设备给出检测线荧光信号强度与质量控制线荧光信号强度的比值输出。将该比值与标本浓度的关系绘制成工作曲线。将我们方法获得的62份临床标本的结果与罗氏E411试剂盒获得的结果进行平行比较。
基于对数 - 对数图,新方法表明在20 pg/mL至10 ng/mL的NT - proBNP浓度范围内,比值与NT - proBNP浓度之间存在良好的线性关系。我们方法检测的62份临床标本的结果与罗氏E411试剂盒检测的结果显示出良好的线性相关性。
基于近红外POCT设备的NT - proBNP水平新LFA检测方法快速、高灵敏度且应用范围广泛,因此适用于心脏损伤的快速早期临床诊断。