Guangling College, School of Chemistry and Chemical Engineering, Yangzhou University, Yangzhou, 225002, PR China.
Department of Laboratory Medicine and Clinical Medical College of Yangzhou University, Subei Peoples' Hospital of Jiangsu Province, Yangzhou, 225001, PR China.
Anal Chim Acta. 2019 Feb 21;1049:213-218. doi: 10.1016/j.aca.2018.10.050. Epub 2018 Oct 25.
Quantitative detection of multiple chicken cytokines is a good evaluation of cell-mediated immunity in chickens after disease infection or vaccination. However, current assay methods for chicken cytokines cannot meet the needs of clinical diagnosis due to unsatisfactory sensitivity and low assay throughput. Herein, a sensitive chemiluminescence (CL) imaging immunosensor array has been developed for high-throughput detection of multiple chicken cytokines. The chicken cytokines immunosensor array was prepared by assembling different cytokine capture antibodies onto a disposable silanized glass chip, where horseradish peroxidase and antibody-conjugated gold nanoparticles were used as multienzymatic amplification probe for CL imaging signal amplification. By using a sandwich assay mode, the amplified CL signals from each sensing array cell were collected for quantitation. Using chicken interleukin-4 and chicken interferon-γ as model cytokines, this novel multiplexed and amplified method demonstrated simultaneous measurement of the two chicken cytokines in the linear ranges of 0.008-0.12 ng/mL and 0.005-0.20 ng/mL, respectively, which yields limits of detection down to 2 pg/mL and 3 pg/mL. The CL imaging array method reported here also demonstrated high specificity, good repeatability, and high stability and accuracy, providing a novel multiplex immunoassay strategy for highly sensitive and high-throughput detection of chicken cytokines and further disease diagnosis in poultry.
定量检测多种鸡细胞因子是评估鸡在疾病感染或接种疫苗后细胞介导免疫的良好方法。然而，由于灵敏度不理想和检测通量低，目前用于鸡细胞因子的检测方法无法满足临床诊断的需求。本文开发了一种灵敏的化学发光（CL）成像免疫传感器阵列，用于高通量检测多种鸡细胞因子。鸡细胞因子免疫传感器阵列通过将不同的细胞因子捕获抗体组装到一次性硅烷化玻璃芯片上制备而成，其中辣根过氧化物酶和抗体偶联的金纳米颗粒被用作 CL 成像信号放大的多酶放大探针。通过使用三明治测定模式，收集每个传感阵列单元的放大 CL 信号进行定量。使用鸡白细胞介素-4 和鸡干扰素-γ 作为模型细胞因子，这种新型的多重和放大方法分别在 0.008-0.12ng/mL 和 0.005-0.20ng/mL 的线性范围内同时测量了这两种鸡细胞因子，检测限低至 2pg/mL 和 3pg/mL。本文报道的 CL 成像阵列方法还表现出高特异性、良好的重复性、高稳定性和准确性，为鸡细胞因子的高灵敏度和高通量检测以及家禽疾病的进一步诊断提供了一种新的多重免疫分析策略。
