Thapa Sandhya, Ghimire Niraj, Chen Fur-Chi
Department of Food and Animal Sciences, Tennessee State University, Nashville, TN 37209, USA.
Department of Human Sciences, Tennessee State University, Nashville, TN 37209, USA.
Microorganisms. 2025 Apr 10;13(4):871. doi: 10.3390/microorganisms13040871.
Many countries have established regulatory frameworks to monitor and mitigate contamination in poultry products. The ability to rapidly quantify is critical for poultry processors to facilitate early detection, implement corrective measures, and enhance product safety. This study aimed to develop an Immunomagnetic Chemiluminescent Assay (IMCA) for the quantification of Typhimurium in ground chicken. Immunomagnetic microbeads functionalized with monoclonal antibodies were employed to selectively capture and concentrate from ground chicken samples. A biotin-labeled monoclonal antibody, followed by an avidin-horseradish peroxidase conjugate, was used to bind the captured bacteria and initiate a chemiluminescent reaction catalyzed by peroxidase. Light emission was quantified in relative light units (RLUs) using two luminometers. Ground chicken samples were inoculated with a four-strain S. Typhimurium cocktail ranging from 0 to 3.5 Log CFU/g. Bacterial concentrations were confirmed using the Most Probable Number (MPN) method. Samples underwent enrichment in Buffered Peptone Water (BPW) supplemented with BAX MP Supplement at 42 °C for 6 and 8 h before analysis via IMCA. A linear regression analysis demonstrated that the optimal quantification of was achieved at the 8 h enrichment period (R ≥ 0.89), as compared to the 6 h enrichment. The limit of quantification (LOQ) was determined to be below 1 CFU/g. A strong positive correlation (R ≥ 0.88) was observed between IMCA and MPN results, indicating methodological consistency. These findings support the application of IMCA as a rapid and reliable method for the detection and quantification of in ground chicken.
许多国家已建立监管框架,以监测和减轻禽肉产品中的污染。快速定量的能力对于家禽加工商促进早期检测、实施纠正措施和提高产品安全性至关重要。本研究旨在开发一种免疫磁化学发光分析法(IMCA),用于定量测定碎鸡肉中的鼠伤寒沙门氏菌。用单克隆抗体功能化的免疫磁微珠用于从碎鸡肉样品中选择性捕获和浓缩目标菌。使用生物素标记的单克隆抗体,随后是抗生物素蛋白-辣根过氧化物酶结合物,来结合捕获的细菌并引发由过氧化物酶催化的化学发光反应。使用两台发光计以相对光单位(RLU)对发光进行定量。向碎鸡肉样品中接种0至3.5 Log CFU/g的四株鼠伤寒沙门氏菌混合菌液。使用最大可能数(MPN)法确认细菌浓度。在通过IMCA分析之前,样品在补充有BAX MP补充剂的缓冲蛋白胨水(BPW)中于42°C富集6小时和8小时。线性回归分析表明,与6小时富集相比,在8小时富集期实现了目标菌的最佳定量(R≥0.89)。定量限(LOQ)确定为低于1 CFU/g。IMCA与MPN结果之间观察到强正相关(R≥0.88),表明方法的一致性。这些发现支持将IMCA作为一种快速可靠的方法用于检测和定量碎鸡肉中的目标菌。