The detection of cytokines plays an important role in clinical diagnosis and immune mechanism research of chicken diseases. In this work, a novel and ultrasensitive chemiluminescent (CL) imaging array immunosensor was proposed to detect multiple chicken cytokines based on DNAzyme@CuS nanoparticles (DNAzyme@CuSNPs) dual mimic enzyme signal amplification strategy. DNAzyme@CuSNPs owns excellent peroxidase property, which was modified with second antibody (Ab₂) to prepare DNAzyme@CuSNPs detection probe, and demonstrated high catalysis CL imaging signal due to synergistic catalysis. Chicken interleukin-4 (ChIL-4) and chicken interferon-γ (ChIFN-γ) were used as model analysis samples, the DNAzyme@CuSNPs-based CL imaging immunosensor achieved simultaneous and high-throughput detection of ChIL-4 and ChIFN-γ with wide linear range of 10⁻³–10² ng/mL, and the detection limits are 0.41 pg/mL and 0.36 pg/mL, respectively. The multiplex chicken cytokines CL imaging array immunosensor shows a high sensitivity, wide linear range, excellent specificity and acceptable stability. This research opens dual mimic enzyme signal-amplified strategy to develop sensitive CL imaging immunoassay for chicken diseases detection application.

细胞因子的检测对于鸡疾病的临床诊断和免疫机制研究具有重要作用。在这项工作中，提出了一种新型超灵敏化学发光（CL）成像阵列免疫传感器，基于DNAzyme@CuS纳米粒子（DNAzyme@CuSNPs）双模拟酶信号放大策略来检测多种鸡细胞因子。DNAzyme@CuSNPs具有优异的过氧化物酶特性，用二抗（Ab ₂）对其进行修饰，制备DNAzyme@CuSNPs检测探针，并由于协同催化而表现出高催化CL成像信号。鸡白介素-4 (ChIL-4) 和鸡干扰素- γ (ChIFN- γ）作为模型分析样品，基于DNAzyme@CuSNPs的CL成像免疫传感器实现了ChIL-4和ChIFN- γ的同时高通量检测，线性范围为10 ^-3 –10 ²  ng/mL，检测结果限值分别为 0.41 pg/mL 和 0.36 pg/mL。多重鸡细胞因子CL成像阵列免疫传感器具有高灵敏度、宽线性范围、优异的特异性和可接受的稳定性。这项研究开启了双模拟酶信号放大策略，开发用于鸡疾病检测应用的灵敏 CL 成像免疫分析。