Zhejiang Provincial Key Laboratory of Biometrology and Inspection & Quarantine, College of Life Sciences, China Jiliang University, Hangzhou, 310018, China.
MOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, 210095, China.
Mikrochim Acta. 2024 Sep 11;191(10):589. doi: 10.1007/s00604-024-06652-2.
Rapid and reliable detection method for African swine fever virus (ASFV) is proposed by surface-enhanced Raman spectroscopy (SERS). The ASFV target DNA can be specifically captured by sandwich hybridization between nanomagnetic beads and a SERS probe. Experimental results show that the significant Raman signal of the SERS probe with gold nanoparticles and a molecular reporter DTNB (5,5'-dimercapto-bis (2-nitrobenzoic acid)) can be adopted for detecting the hybridization chain reaction of ASFV DNA. The advantage of the SERS sandwich hybridization assay is the large response range from the single molecule level to 10 copies per mL, which not only can overcome the tedious time required for the amplification reaction but also provides a comparative method to polymerase chain reaction. Furthermore, real samples of African swine fever virus were detected from different subjects of swine fever virus including porcine reproductive respiratory syndrome virus and Japanese encephalitis virus. The proposed biosensor method can rapidly detect ASFV correctly within 15 min as a simple, convenient, low-cost detection approach. The biosensor can be used as a platform for the determination in biological, food, and environmental analytical fields.
提出了一种基于表面增强拉曼光谱(SERS)的快速、可靠的非洲猪瘟病毒(ASFV)检测方法。纳米磁珠与 SERS 探针之间的夹心杂交可以特异性捕获 ASFV 靶 DNA。实验结果表明,采用金纳米粒子和分子报告剂 DTNB(5,5'-二巯基-双(2-硝基苯甲酸))的 SERS 探针具有显著的拉曼信号,可用于检测 ASFV DNA 的杂交链反应。SERS 夹心杂交测定法的优点是从单分子水平到 10 拷贝/mL 的大响应范围,不仅可以克服扩增反应所需的繁琐时间,还提供了与聚合酶链反应相比的方法。此外,还从猪繁殖与呼吸综合征病毒和日本脑炎病毒等不同猪瘟病毒的对象中检测到了非洲猪瘟病毒的实际样本。该生物传感器方法可在 15 分钟内快速准确地检测 ASFV,是一种简单、方便、低成本的检测方法。该生物传感器可作为生物、食品和环境分析领域中测定的平台。