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Ultrasensitive chemiluminescence immunoassay with enhanced precision for the detection of cTnI amplified by acridinium ester-loaded microspheres and internally calibrated by magnetic fluorescent nanoparticles
Nanoscale ( IF 5.8 ) Pub Date : 2021-1-19 , DOI: 10.1039/d0nr08008j Huan Zhao 1, 2, 3, 4, 5 , Qifeng Lin 6, 7, 8, 9 , Li Huang 1, 2, 3, 4, 5 , Yunfeng Zhai 1, 2, 3, 4, 5 , Yuan Liu 1, 2, 3, 4, 5 , Yan Deng 10, 11, 12, 13 , Enben Su 1, 2, 3, 4, 5 , Nongyue He 1, 2, 3, 4, 5
Nanoscale ( IF 5.8 ) Pub Date : 2021-1-19 , DOI: 10.1039/d0nr08008j Huan Zhao 1, 2, 3, 4, 5 , Qifeng Lin 6, 7, 8, 9 , Li Huang 1, 2, 3, 4, 5 , Yunfeng Zhai 1, 2, 3, 4, 5 , Yuan Liu 1, 2, 3, 4, 5 , Yan Deng 10, 11, 12, 13 , Enben Su 1, 2, 3, 4, 5 , Nongyue He 1, 2, 3, 4, 5
Affiliation
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A novel enhanced chemiluminescent immunoassay (CLIA) for ultrasensitive and excellent precisive determination of cardiac troponin I (cTnI) was reported. The method made full use of poly[(N-isopropyl acrylamide)-co-(methacrylic acid)] (P(NIPAM-co-MAA)) microspheres as new potential signal enhancers and magnetic fluorescent nanoparticles as internal standards for better precision. This protocol involved a sandwich format, in which the antigen in the sample was captured by the immobilized antibodies on the surface of magnetic fluorescent beads and recognized by the other antibodies labeled with acridinium ester (AE)-loaded P(NIPAM-co-MAA) microspheres. The combination of the remarkable sensitivity of the enhanced CLIA method and the use of P(NIPAM-co-MAA) microspheres as anti-cTnI carriers for acridinium ester signal amplification provided an extremely sensitive limit of blank (LoB) at 0.097 pg mL−1, a limit of detection (LoD) at 0.116 pg mL−1, and a limit of quantitation (LoQ) at 0.606 pg mL−1, much greater than those achieved by the classical chemiluminescence immunoassay (CLIA, Getein). Moreover, the intra-day variable coefficient can be improved to 1.21–2.12%, and inter-day variability was 2.01–3.49% under the application of magnetic fluorescent beads as an internal standard. The sensitivity and precision have reached a high level, comparable with the current commercial detection kits. The results showed a good correlation with a commercial chemiluminescence assay (CLIA, Abbott), with a correlation coefficient of 0.9883. This proposed method has been successfully applied to the clinical determination of cTnI in the human serum.
中文翻译：
超灵敏的化学发光免疫分析法，具有更高的精密度，可检测载有cri啶酯的微球扩增的cTnI，并通过磁性荧光纳米粒子进行内部校准
报道了一种新颖的增强型化学发光免疫分析法（CLIA），用于心脏肌钙蛋白I（cTnI）的超灵敏和精确测定。该方法充分利用了聚[（N-异丙基丙烯酰胺）-co-（甲基丙烯酸）]（P（NIPAM- co- MAA））微球作为新的潜在信号增强剂，并利用磁性荧光纳米粒子作为内标，以提高精度。该方案涉及一种三明治形式，其中样品中的抗原被固定在磁性荧光珠表面的抗体捕获，并被载有cri啶酯（AE）的P（NIPAM- co）标记的其他抗体识别-MAA）微球。增强的CLIA方法的出色灵敏度与使用P（NIPAM- co -MAA）微球作为抗cTnI载体进行a啶酯信号放大的结合提供了0.097 pg mL -1的空白（LoB）极高灵敏度极限，检测限（LoD）为0.116 pg mL -1和定量限（LoQ）为0.606 pg mL -1，比经典的化学发光免疫分析（CLIA，Getein）所获得的结果要大得多。此外，在使用磁性荧光珠作为内标的情况下，日内可变系数可以提高到1.21-2.12％，日间可变性是2.01-3.49％。与目前的商用检测套件相比，其灵敏度和精度已达到很高的水平。结果显示与商业化学发光分析（CLIA，Abbott）具有良好的相关性，相关系数为0.9883。该方法已成功应用于人血清中cTnI的临床测定。
更新日期：2021-02-03
中文翻译：
超灵敏的化学发光免疫分析法，具有更高的精密度，可检测载有cri啶酯的微球扩增的cTnI，并通过磁性荧光纳米粒子进行内部校准
报道了一种新颖的增强型化学发光免疫分析法（CLIA），用于心脏肌钙蛋白I（cTnI）的超灵敏和精确测定。该方法充分利用了聚[（N-异丙基丙烯酰胺）-co-（甲基丙烯酸）]（P（NIPAM- co- MAA））微球作为新的潜在信号增强剂，并利用磁性荧光纳米粒子作为内标，以提高精度。该方案涉及一种三明治形式，其中样品中的抗原被固定在磁性荧光珠表面的抗体捕获，并被载有cri啶酯（AE）的P（NIPAM- co）标记的其他抗体识别-MAA）微球。增强的CLIA方法的出色灵敏度与使用P（NIPAM- co -MAA）微球作为抗cTnI载体进行a啶酯信号放大的结合提供了0.097 pg mL -1的空白（LoB）极高灵敏度极限，检测限（LoD）为0.116 pg mL -1和定量限（LoQ）为0.606 pg mL -1，比经典的化学发光免疫分析（CLIA，Getein）所获得的结果要大得多。此外，在使用磁性荧光珠作为内标的情况下，日内可变系数可以提高到1.21-2.12％，日间可变性是2.01-3.49％。与目前的商用检测套件相比，其灵敏度和精度已达到很高的水平。结果显示与商业化学发光分析（CLIA，Abbott）具有良好的相关性，相关系数为0.9883。该方法已成功应用于人血清中cTnI的临床测定。
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