Rapid detection of Salmonella in food and feed by coupling loop-mediated isothermal amplification with bioluminescent assay in real-time.
Rapid detection of Salmonella in food and feed by coupling loop-mediated isothermal amplification with bioluminescent assay in real-time.
复制标题
通过将环介导的等温扩增与实时生物发光检测相结合，快速检测食品和饲料中的沙门氏菌。
DOI：
10.1186/s12866-016-0730-7
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发表时间：
2016-06-17
期刊：
影响因子：
4.2
通讯作者：
Ge B
中科院分区：
文献类型：
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作者：
Yang Q;Domesle KJ;Wang F;Ge B
沙门氏菌是引起食品和饲料安全问题的最重要病原体之一。本研究探讨了环介导等温扩增（LAMP）与实时生物发光法（BART）耦合快速检测各种食品和饲料样品中沙门氏菌的方法。将LAMP- bart分析的性能与传统LAMP和市售的3M沙门氏菌分子检测分析（MDA）进行比较。LAMP-BART法对178株（151株沙门菌和27株非沙门菌）具有100%的特异性。检测限在纯培养条件下为36个细胞/反应，在未富集的加标食品和饲料样品中为104 ~ 106个CFU / 25 g，与传统LAMP和3M MDA沙门氏菌的检测限相当，但速度快5 ~ 10 min。地面火鸡对3M MDA沙门氏菌有很强的抑制作用，每25 g至少需要108 CFU才能检测到。沙门氏菌细胞数与LAMP-BART信号具有较高的相关性（R2 = 0.941 ~ 0.962），具有较好的定量能力。富集24小时后，在五种食物（哈密瓜、碎牛肉、碎火鸡、壳蛋和番茄）和三种饲料（牛饲料、鸡饲料和干狗粮）中，所有三种检测方法均准确检测到每25 g沙门氏菌中含有1至3 CFU。然而，当用3M MDA沙门氏菌检测时，每25克牛饲料需要101 CFU。沙门氏菌LAMP-BART检测具有快速、特异、灵敏、定量、稳健性好等特点。在进一步验证后，它可能成为在各种类型的食品和饲料样品中进行沙门氏菌常规筛选的有价值的工具。
Salmonella is among the most significant pathogens causing food and feed safety concerns. This study examined the rapid detection of Salmonella in various types of food and feed samples by coupling loop-mediated isothermal amplification (LAMP) with a novel reporter, bioluminescent assay in real-time (BART). Performance of the LAMP-BART assay was compared to a conventional LAMP and the commercially available 3M Molecular Detection Assay (MDA) Salmonella. The LAMP-BART assay was 100 % specific among 178 strains (151 Salmonella and 27 non-Salmonella) tested. The detection limits were 36 cells per reaction in pure culture and 104 to 106 CFU per 25 g in spiked food and feed samples without enrichment, which were comparable to those of the conventional LAMP and 3M MDA Salmonella but 5–10 min faster. Ground turkey showed a strong inhibition on 3M MDA Salmonella, requiring at least 108 CFU per 25 g for detection. The correlation between Salmonella cell numbers and LAMP-BART signals was high (R2 = 0.941–0.962), suggesting good quantification capability. After 24 h enrichment, all three assays accurately detected 1 to 3 CFU per 25 g of Salmonella among five types of food (cantaloupe, ground beef, ground turkey, shell eggs, and tomato) and three types of feed (cattle feed, chicken feed, and dry dog food) examined. However, 101 CFU per 25 g was required for cattle feed when tested by 3M MDA Salmonella. The Salmonella LAMP-BART assay was rapid, specific, sensitive, quantitative, and robust. Upon further validation, it may become a valuable tool for routine screening of Salmonella in various types of food and feed samples.