National Research Laboratory of Molecular Biotechnology, Department of Chemical Engineering, Pohang University of Science and Technology, Pohang 790-784, South Korea.
Appl Biochem Biotechnol. 2010 Oct;162(4):1187-94. doi: 10.1007/s12010-009-8853-z. Epub 2009 Nov 24.
The polymerase chain reaction (PCR) technique is widely used for efficient detection of food-borne pathogens because of speed and specificity. However, PCR methods have focused mostly on species-specific detection. In the present work, we describe a PCR-based method for the simultaneous detection of all Vibrio species because lots of them are notorious food-borne human pathogens. We then combined this total detection method with specific detection of Vibrio cholerae pathogen. Using a degenerate primer set based on the sequence of the potassium uptake gene, trkA, we were able to successfully detect all Vibrio species. Specific detection of V. cholerae was also possible using primer sets based on putative flagellin sequence. Importantly, simultaneous total and species-specific Vibrio detection was possible using all two primer sets in a multiplexed PCR strategy. Thus, the PCR method we have developed is applicable to both simultaneous and two-step detection of total and specific Vibrio species.
聚合酶链反应（PCR）技术因其速度和特异性而被广泛用于高效检测食源性病原体。然而，PCR 方法主要集中于物种特异性检测。在本工作中，我们描述了一种基于 PCR 的方法，用于同时检测所有弧菌属物种，因为它们中的许多都是臭名昭著的食源性病原体。然后，我们将这种总检测方法与霍乱弧菌病原体的特异性检测相结合。使用基于钾摄取基因 trkA 序列的简并引物对，我们成功地检测到所有弧菌属物种。使用基于假定鞭毛序列的引物对也可以特异性检测霍乱弧菌。重要的是，使用这两个引物对的多重 PCR 策略可以同时进行总菌和种特异性弧菌的总菌和种特异性检测。因此，我们开发的 PCR 方法可同时适用于总菌和种特异性弧菌的两步法和两步法检测。
