This study reports the development and optimization of a real-time loop-mediated isothermal amplification (qLAMP) method for rapid detection of Acetobacter aceti strain in red wine samples. Our results showed that the primers and probes designed for 16S rRNA were effective for A. aceti detection. The quantification limit of real-time polymerase chain reaction (qPCR) and qLAMP in pure culture was 2.05 × 10¹ colony forming units (CFU) mL⁻¹. qLAMP had a sensitivity of 6.88 × 10¹ CFU mL⁻¹ in artificially contaminated Changyu dry red wine (CDRW) and Changyu red wine (CRW), and 6.88 × 10² CFU mL⁻¹ in artificially contaminated Greatwall dry red wine (GDRW), which was 10 times higher than that of qPCR. In conclusion, this newly developed qLAMP is a reliable, rapid and accurate method for the detection and quantification of A. aceti species in red wine samples. Furthermore, our work provides a standard reference method for the quantitative detection of A. aceti and other acetic acid bacteria during the fermentation and storage of red wine samples.

中文翻译：

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实时环介导的等温扩增定量红酒中乙醋的方法的开发与应用

这项研究报告了实时循环介导的等温扩增（qLAMP）方法的开发和优化，该方法可快速检测红酒样品中的醋杆菌。我们的结果表明，设计用于16S rRNA的引物和探针可有效用于乙酰丙酮杆菌的检测。纯培养液中实时聚合酶链反应（qPCR）和qLAMP的定量限为2.05×10 ¹集落形成单位（CFU）mL ^-1。在人工污染的张裕干红葡萄酒（CDRW）和张裕红葡萄酒（CRW）中，qLAMP的灵敏度为6.88×10 ¹ CFU mL ^-1，灵敏度为6.88×10 ² CFU mL ^-1人为污染的长城干红葡萄酒（GDRW）中的含量是qPCR的10倍。总而言之，这种新开发的qLAMP是一种可靠，快速且准确的方法，用于检测和定量红酒样品中的醋曲菌。此外，我们的工作为在红酒样品的发酵和储存过程中定量检测乙酰乙酸杆菌和其他乙酸细菌提供了一种标准的参考方法。