Institute of Biochemistry and Biophysics, University of Tehran, P. O. Box 13145-1384, Tehran, Iran.
World J Microbiol Biotechnol. 2012 May;28(5):2101-6. doi: 10.1007/s11274-012-1014-5. Epub 2012 Feb 9.
Loop-mediated isothermal amplification (LAMP) is a promising nucleic acid assay for rapid and cost-effective detection of pathogen-specific sequences within a sample. Development of an appropriate taxonomic group-specific LAMP assay highly relies on the design of proper primers to cover all major members of the taxon. Regarding this fact, we designed and evaluated a new LAMP primer set specific to prt (rfbS) gene for rapid identification of Salmonella serogroup D serotypes. Unlike the previously reported LAMP assay for serogroup D which detects solely the non-typhoidal serotypes; the new LAMP primers set detects both typhoidal and non-typhoidal serotypes of this serogroup with a detection limit of 10 CFU/rection. Furthermore, the technique was successfully applied to artificially contaminated meat samples with an inoculation level of 1-5 CFU/250 ml of Salmonella Enteritidis, following a 5-h pre-enrichment step in tryptic soy broth. Overall, the new LAMP assay and its optimized setup would be useful for fast diagnosis of food poisoning incidents caused by these bacteria.
环介导等温扩增（LAMP）是一种很有前途的核酸检测方法，可用于快速、经济地检测样本中病原体特异性序列。开发适当的分类群特异性 LAMP 检测方法高度依赖于设计适当的引物以覆盖该分类群的所有主要成员。关于这一事实，我们设计并评估了一种新的针对 prt（rfbS）基因的 LAMP 引物组，用于快速鉴定沙门氏菌血清群 D 血清型。与之前报道的仅检测非伤寒型血清型的血清群 D 的 LAMP 检测方法不同；新的 LAMP 引物组可检测该血清群的伤寒型和非伤寒型血清型，检测限为 10 CFU/反应。此外，该技术在经过 5 小时的胰蛋白酶大豆肉汤预增菌后，成功应用于接种水平为 1-5 CFU/250 ml 的肠炎沙门氏菌人工污染的肉样中。总体而言，这种新的 LAMP 检测方法及其优化设置可用于快速诊断由这些细菌引起的食物中毒事件。
