The loop-mediated isothermal amplification (LAMP) technique was used to investigate six salmonella-specific sequences for their suitability to serve as targets for the pathogen identification. Sequences selected for designing LAMP primers were genes invA, bcfD, phoP, siiA, gene62181533 and a region within the ttrRSBCA locus. Primers including single nucleotide polymorphisms were configured as degenerate primers. Specificity of the designed primer sets was determined by means of 46 salmonella and 26 non-salmonella strains. Primers targeting the ttrRSBCA locus showed 100 % inclusivity and exclusivity of the tested target and non-target strains, respectively. Other primer sets revealed deficiencies especially regarding Salmonella enterica subsp. II - IV and Salmonella bongori. Additionally, primers targeting the siiA gene failed to detect Salmonella enterica serotypes Newport and Stanley, whereas bcfD primers did not amplify DNA of Salmonella enterica serotype Schleissheim. TtrRSBCA primers, providing short detection times and constant melting temperatures of amplificates, achieved best overall performance.

中文翻译：

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不同靶基因检测沙门氏菌的评价。环介导等温扩增（LAMP）

环介导的等温扩增 (LAMP) 技术用于研究六个沙门氏菌特异性序列，以确定它们是否适合作为病原体鉴定的目标。选择用于设计 LAMP 引物的序列是基因 invA、bcfD、phoP、siiA、gene62181533 和 ttrRSBCA 基因座内的一个区域。包括单核苷酸多态性的引物被配置为简并引物。设计的引物组的特异性通过 46 种沙门氏菌和 26 种非沙门氏菌菌株确定。靶向 ttrRSBCA 基因座的引物分别显示出 100% 的测试目标和非目标菌株的包容性和排他性。其他引物组揭示了缺陷，尤其是关于沙门氏菌亚种。II - IV 和沙门氏菌 bongori。此外，针对 siiA 基因的引物未能检测到沙门氏菌血清型 Newport 和 Stanley，而 bcfD 引物不能扩增肠道沙门氏菌血清型 Schleissheim 的 DNA。TtrRSBCA 引物提供较短的检测时间和恒定的扩增产物解链温度，实现了最佳的整体性能。