| Abstrakt: |
Abstract: Identification of extremely small amounts of the bacterial contaminants in water and food has always been the main challenge facing scientists and researchers. To overcome this challenge, bacteria culture is considered the only conventional method available for bacteria enrichment and detection. But this procedure is highly time-consuming and requires long-term incubation. However, by employing such non-culture, intelligent, and newly developed methods as immuno-magnetic separation (IMS), bacteria can be trapped with relative ease, thus reducing the time needed for the detection process. The present research work aims to culture enrich Vibrio cholerae, one of the most important contaminators of water supplies, using the IMS method. The obtained results are then utilized for comparative and functional evaluation with those acquired through the three common methods of the bacteria detection process; cell culture, ATP assessment, and PCR. For this, a specified concentration of the bacteria was dispersed in 1 mL of iron oxide (Fe3O4) NPs coated with arginine and lysine. The Result before and after applying the IMS method was compared with cell culture, ATP assessment, and molecular detection (PCR) with specific primers (ompWgene) of V. cholera.The data showed the high efficiency and adequate performance of the proposed method. According to the ATP assay, the sensitive detection of Fe3O4@Arg and Fe3O4@Lys NPs were determined to be about 2 × 105and 2 × 102CFU, respectively. In addition, according to PCR, the sensitivity detection of Fe3O4@Arg and Fe3O4@Lys NPs were determined about 3.2 × 105and 3.2 × 102CFU, respectively. Accordingly, V. choleraecould be isolated and enriched in less than 60 min, and using the molecular PCR method it could be detected in less than 4 h. |
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