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用可见/近红外高光谱显微镜成像技术检测鸡冲洗液中的沙门氏菌与 RT-PCR 的比较。

Detection of Salmonella from chicken rinsate with visible/near-infrared hyperspectral microscope imaging compared against RT-PCR.

机构信息

US Department of Agriculture, Agricultural Research Services, US National Poultry Research Center, Athens, GA 30605, USA.

US Department of Agriculture, Agricultural Research Services, US National Poultry Research Center, Athens, GA 30605, USA.

出版信息

Talanta. 2019 Apr 1;195:313-319. doi: 10.1016/j.talanta.2018.11.071. Epub 2018 Nov 23.

DOI:10.1016/j.talanta.2018.11.071
PMID:30625548
Abstract

Salmonella is an organism of importance to the poultry industry with increasingly stringent government regulatory standards. Real-time polymerase chain reaction (RT-PCR) and plating procedures on nutrient enriched growth media have been the standard detection methods of Salmonella from broiler chicken carcasses for years. These methods are proven, but offer disadvantages in the amount of time or reoccurring sample cost. Here, we propose the use of a hyperspectral microscope imaging system (HMI) for comparison to standard detection methods. Broiler chicken carcasses were rinsed and plated on Salmonella selective agar. Colonies from plates were picked and RT-PCR was used as a confirmation test to verify plating results, while HMI was collected from the same colonies. Spectral signatures of cells were extracted between 450 and 800 nm from HMI collected with 100x objective. A quadratic discriminant analysis (QDA) was used to classify cells as either Salmonella positive or negative (n = 341). Spectra preprocessing minimized the influence of cellular shape on the spectra, increasing the initial classification accuracy of 81.8-98.5%, yielding a sensitivity of 1.0, and a specificity of 0.963. Results showed the potential as an initial investigation of HMI as a microbial confirmation tool, compared to RT-PCR.

摘要

沙门氏菌是家禽业的重要生物体,政府监管标准越来越严格。实时聚合酶链反应(RT-PCR)和营养丰富的生长培养基上的平板程序多年来一直是从肉鸡胴体中检测沙门氏菌的标准检测方法。这些方法是经过验证的,但在时间或重复样本成本方面存在缺点。在这里,我们提出使用高光谱显微镜成像系统(HMI)与标准检测方法进行比较。将肉鸡胴体冲洗并在选择性沙门氏菌琼脂平板上进行平板划线。从平板上挑取菌落,并使用 RT-PCR 作为确认试验来验证平板划线结果,同时从相同的菌落收集 HMI。从用 100x 物镜收集的 HMI 中提取 450 到 800nm 之间的细胞光谱特征。使用二次判别分析(QDA)将细胞分类为沙门氏菌阳性或阴性(n=341)。光谱预处理最小化了细胞形状对光谱的影响,初始分类准确率从 81.8%提高到 98.5%,灵敏度为 1.0,特异性为 0.963。结果表明,与 RT-PCR 相比,HMI 作为微生物确认工具具有初步研究的潜力。

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