比较传统 PCR、多重 PCR 和环介导等温扩增检测方法对弯曲杆菌种的快速检测。
Comparison of conventional PCR, multiplex PCR, and loop-mediated isothermal amplification assays for rapid detection of Arcobacter species.
机构信息
Department of Food and Nutrition, School of Food Science and Technology, Chung-Ang University, Ansung-Si, Gyeonggi-Do, South Korea.
出版信息
J Clin Microbiol. 2014 Feb;52(2):557-63. doi: 10.1128/JCM.02883-13. Epub 2013 Dec 11.
Abstract
This study aimed to develop a loop-mediated isothermal amplification (LAMP) method for the rapid detection of Arcobacter species. Specific primers targeting the 23S ribosomal RNA gene were used to detect Arcobacter butzleri, Arcobacter cryaerophilus, and Arcobacter skirrowii. The specificity of the LAMP primer set was assessed using DNA samples from a panel of Arcobacter and Campylobacter species, and the sensitivity was determined using serial dilutions of Arcobacter species cultures. LAMP showed a 10- to 1,000-fold-higher sensitivity than multiplex PCR, with a detection limit of 2 to 20 CFU per reaction in vitro. Whereas multiplex PCR showed cross-reactivity with Campylobacter species, the LAMP method developed in this study was more sensitive and reliable than conventional PCR or multiplex PCR for the detection of Arcobacter species.
摘要
本研究旨在开发一种环介导等温扩增（LAMP）方法，用于快速检测弯曲杆菌属物种。使用针对 23S 核糖体 RNA 基因的特异性引物来检测弯曲杆菌属、嗜冷弯曲杆菌和斯基罗弯曲杆菌。通过使用来自一组弯曲杆菌属和空肠弯曲菌属物种的 DNA 样本评估 LAMP 引物组的特异性，并通过系列稀释弯曲杆菌属培养物来确定敏感性。LAMP 的灵敏度比多重 PCR 高 10 到 1000 倍，在体外反应中检测限为 2 到 20 CFU/反应。虽然多重 PCR 与空肠弯曲菌属物种有交叉反应，但本研究中开发的 LAMP 方法比传统 PCR 或多重 PCR 更敏感和可靠，可用于检测弯曲杆菌属物种。
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