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Quantification of Viable Salmonella by Propidium Monoazide Real-Time PCR After Long-Term Storage of Peanut Products

机译:花生制品长期储存后通过单叠氮化丙啶实时 PCR 定量分析活沙门氏菌

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摘要

In this study, the performance of quantitative PCR, combined or not with propidium monoazide (PMA), to recover Salmonella from peanut products after different storage times was evaluated. The samples were inoculated with 5–6 log cfu g−1 of Salmonella Typhimurium ATCC 14028 and stored at 28 °C for up to 540 d. The correlation between the threshold cycle number (Ct) and the colony-forming units (cfu) was obtained by a standard curve, which showed a linear correlation (R2 = 0.97). The highest counts were recovered by qPCR (p < 0.05); however, it quantified both viable and non-viable cells. For roasted peanuts, a significant difference (p < 0.05) between qPCR-PMA and the culture method was verified only for samples stored for 30 d, i.e., 2.8 versus 4.0 log cfu g−1. Further, there was no VBNC status in the roasted peanuts, even after long-term exposure to desiccation stress. For peanut-based products, after 540 d, only paçoca showed a significant difference (p < 0.05) among the three methods evaluated. In peanut brittle, qPCR-PMA detected 1.5 log cfu g−1, while, in the culture method, Salmonella was recovered in 1 g. The pathogen was below the detection limit in pé-de-moça either by plate count or qPCR-PMA. Therefore, qPCR-PMA shows potential for use in quantifying Salmonella in peanut products.
机译:在本研究中,评估了定量 PCR 与单叠氮化丙啶 (PMA) 联合或不联合使用在不同储存时间后从花生产品中回收沙门氏菌的性能。用 5-6 log cfu g-1 的鼠伤寒沙门氏菌 ATCC 14028 接种样品,并在 28 °C 下储存长达 540 天。阈值循环数 (Ct) 和菌落形成单位 (cfu) 之间的相关性通过标准曲线获得,该曲线显示线性相关 (R2 = 0.97)。通过 qPCR 回收计数最高 (p < 0.05);然而,它量化了活细胞和非活细胞。对于烤花生,qPCR-PMA 和培养方法之间的显着差异 (p < 0.05) 仅在储存 30 天的样品中得到验证,即 2.8 与 4.0 log cfu g-1。此外,即使在长期暴露于干燥胁迫后,烤花生也没有 VBNC 状态。对于以花生为基础的产品,在 540 天后,只有 paçoca 在评估的三种方法中显示出显着差异 (p < 0.05)。在花生脆中,qPCR-PMA 检测到 1.5 log cfu g-1,而在培养方法中,沙门氏菌回收率为 1 g。病原体在pé-de-moça中通过平板计数或qPCR-PMA均低于检测限。因此,qPCR-PMA 显示出用于定量花生产品中沙门氏菌的潜力。

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