Development of a real-time nucleic acid sequence–based amplification assay for the rapid detection of Salmonella spp. from food
Development of a real-time nucleic acid sequence–based amplification assay for the rapid detection of Salmonella spp. from food
复制标题
开发基于实时核酸序列的扩增技术，用于快速检测食品中的沙门氏菌。
DOI：
10.1007/s42770-018-0002-9
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发表时间：
2018
影响因子：
2.2
通讯作者：
X. Bie
中科院分区：
文献类型：
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作者：
Ligong Zhai;Hongxia Liu;Qiming Chen;Zhaoxin Lu;Chong Zhang;F. Lv;X. Bie
沙门氏菌是最常见的食源性传染病病原体之一。本研究旨在建立食品中沙门氏菌的实时核酸序列扩增（NASBA）检测方法。设计针对沙门氏菌特异性xcd基因的引物和分子信标，对48株沙门氏菌和18株非沙门氏菌进行了检测。经12 h预富集后，该方法对沙门氏菌的特异性高，检出限低（7 × 10−1CFU/mL）。重要的是，它可以检测活细胞。此外，在存在猪肉背景微生物群的情况下检查了NASBA测定法的有效性;它可以检测9.5 × 103 CFU/mL的沙门氏菌细胞。最后，将该方法成功应用于猪肉、牛肉和牛奶中沙门氏菌的检测，检测限可达10 CFU/25 g（mL）。本研究建立的实时NASBA检测方法可用于动物源性食品中沙门氏菌的快速、特异、灵敏检测。
Salmonellaspp. is one of the most common foodborne infectious pathogen. This study aimed to develop a real-time nucleic acid sequence–based amplification (NASBA) assay for detectingSalmonellain foods. Primers and a molecular beacon targeting theSalmonella-specificxcdgene were designed for mRNA transcription, and 48Salmonellaand 18 non-Salmonellastrains were examined. The assay showed a high specificity and low detection limit forSalmonella(7 × 10−1CFU/mL) after 12 h of pre-enrichment. Importantly, it could detect viable cells. Additionally, the efficacy of the NASBA assay was examined in the presence of pork background microbiota; it could detectSalmonellacells at 9.5 × 103CFU/mL. Lastly, it was successfully used to detectSalmonellain pork, beef, and milk, and its detection limit was as low as 10 CFU/25 g (mL). The real-time NASBA assay developed in this study may be useful for rapid, specific, and sensitive detection ofSalmonellain food of animal origin.