Fujian Key Laboratory of Drug Target Discovery and Structural and Functional Research, School of Pharmacy, Fujian Medical University, Fuzhou, 350004, China.
Department of Pharmacy, Affiliated Quanzhou First Hospital of Fujian Medical University, Quanzhou, 362000, China.
Mikrochim Acta. 2022 Mar 26;189(4):160. doi: 10.1007/s00604-022-05263-z.
Sensitive and rapid detection of pathogenic bacteria plays an important role in avoiding food poisoning. However, the practical application value of conventional assays for detection of foodborne bacteria, are limited by major drawbacks; these include the laboriousness of pure culture preparation, complexity of DNA extraction for polymerase chain reaction, and low sensitivity of enzyme-linked immunosorbent assay. Herein, we designed a non-complex strategy for the sensitive, quantitative, and rapid detection of Salmonella typhimurium with high specificity, using an anti-Salmonella typhimurium IgG-AuNC-based immunofluorescent-aggregation assay. Salmonella typhimurium was agglutinated with fluorescent anti-Salmonella typhimurium IgG-AuNC on a glass slide, and observed using a fluorescence microscope with photoexcitation and photoemission at 560 nm and 620 nm, respectively. Under optimized reaction conditions, the AuNC-based immunofluorescent-aggregation assay had a determination range between 7.0 × 10 and 3.0 × 10 CFU/mL, a limit of detection of 1.0 × 10 CFU/mL and an assay response time of 3 min. The technique delivered good results in assessing real samples.
灵敏、快速检测致病菌对于避免食物中毒至关重要。然而，传统的食源性细菌检测方法在实际应用中存在局限性，主要缺点包括：纯培养物制备费力、聚合酶链反应的 DNA 提取复杂以及酶联免疫吸附测定的灵敏度低。在此，我们设计了一种基于抗鼠伤寒沙门氏菌 IgG-AuNC 的免疫荧光聚集测定法，用于灵敏、定量和快速检测鼠伤寒沙门氏菌，具有高特异性。在玻片上，鼠伤寒沙门氏菌与荧光标记的抗鼠伤寒沙门氏菌 IgG-AuNC 发生聚集，并分别在 560nm 和 620nm 下用荧光显微镜进行光激发和光发射观察。在优化的反应条件下，基于 AuNC 的免疫荧光聚集测定法的测定范围在 7.0×10 和 3.0×10 CFU/mL 之间，检测限为 1.0×10 CFU/mL，测定响应时间为 3 分钟。该技术在评估实际样品时取得了良好的效果。
