Detection of Vibrio parahaemolyticus in food samples using in situ loop-mediated isothermal amplification method
Detection of Vibrio parahaemolyticus in food samples using in situ loop-mediated isothermal amplification method
复制标题
原位环介导等温扩增法检测食品样品中的副溶血弧菌
DOI：
10.1016/j.gene.2012.12.039
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发表时间：
2013-02-25
期刊：
影响因子：
3.5
通讯作者：
Zhong, Qingping
中科院分区：
文献类型：
--
作者：
Wang, Li;Shi, Lei;Zhong, Qingping
本研究建立了一种新的原位环介导等温扩增（in situ LAMP）技术，用于快速检测食源性副溶血性弧菌菌株。通过对一系列10倍稀释副溶血性弧菌细胞的检测，原位LAMP法的灵敏度为10 CFU/反应，并通过对14株革兰氏阳性和阴性对照菌株的验证，获得了较高的特异性。将建立的原位LAMP法应用于先前从海产品样品中分离的58株，其中48株为副溶血性弧菌，10株为非溶血性弧菌。parahaemolyticus菌株。48株副溶血性弧菌原位LAMP法、常规LAMP法和PCR法检测阳性分别为48株、45株和34株，检出率和阴性预测值分别为100%、93.8%、70.8%和100%、76.9%、41.7%。此外，所有测试的非v。副溶血性链球菌均为阳性，3项检测均为100%阳性预测值（PPV）。与常规LAMP法和基于pcr的方法相比，原位LAMP法具有快速、特异性高、耗时短、操作简便等优点，可为食品安全实验室的病原菌检测提供一种新颖、实用的检测平台。(C) 2012 Elsevier BM。版权所有。
A novel in situ loop-mediated isothermal amplification (in situ LAMP) technique for rapid detection of the food-borne Vibrio parahaemolyticus strains had been developed and evaluated in this study. The sensitivity of the in situ LAMP assay was detected to be 10 CFU/reaction via test in serial 10-fold dilutions of V. parahaemolyticus cells, and high specificity had also been obtained through confirmation with 14 reference gram-positive and -negative strains. Application of the established in situ LAMP assay had been performed on 58 strains previously isolated from seafood samples, including 48 V. parahaemolyticus and 10 non-V. parahaemolyticus strains. Of 48 V. parahaemolyticus strains, 48, 45 and 34 strains were detected as positive by in situ LAMP, regular LAMP and PCR, respectively, with the detection rate and negative predictive value (NPV) found to be 100% vs 93.8% vs 70.8% and 100% vs 76.9% vs 41.7%. In addition, none of the tested non-V. parahaemolyticus strains showed positive result, indicating a 100% positive predictive value (PPV) for all of 3 assays. Compared with regular LAMP methods and PCR-based methods, the in situ LAMP assay is advantageous on rapidity, high specificity, less time consumption and ease in operation, and may provide a novel, useful and practical detection platform for pathogens in food safety laboratories. (C) 2012 Elsevier BM. All rights reserved.