In addition, 1 bacteremia-related strain (strain 11548) was further sequenced by Nanopore MinION (Oxford Nanopore Technologies) to obtain the finished genome sequence. Draft genome sequences were obtained by assembling Illumina reads using Platanus version 1.1.4 with default parameters. Sequencing libraries were prepared using the Nextera XT DNA Sample Prep Kit (Illumina) to obtain paired-end sequences (300 bp × 2) on the Illumina MiSeq platform. Genomic DNA was purified from 2 mL overnight cultures using a DNeasy Blood and Tissue Kit (Qiagen) according to the manufacturer's instructions. After PCR amplification, 10 µL of each PCR product was analyzed by agarose gel electrophoresis using 1.2% agarose S (Nippon Gene). The PCR amplification steps employed were as follows: 25 cycles of initial denaturation at 94☌ for 5 minutes, followed by 40 cycles of denaturation at 94☌ for 1 minute, annealing at 35☌ for 1 minute, and extension at 72☌ for 2 minutes and a final extension step at 72☌ for 10 minutes. Each reaction mixture (25 μL) contained 5 µL of 5× KAPATaq EXtra buffer, 2.5 mM MgCl 2, 300 mM dNTPs, 1 μL of template DNA, 0.4 μM primer, and 0.6 U of KAPATaq DNA polymerase. RAPD polymerase chain reaction (PCR) was performed as previously described by Hashemi et al using 3 primers, namely, OPP-16 (5′-CCAAGCTGCC-3′), OPS-11 (5′-AGTCGGGTGG-3′), and P1254 (5′-CCGCAGCCAA-3′), in conjunction with KAPATaq EXtra (NIPPON Genetics Co, Ltd). Template genomic DNA was prepared by an alkaline boiling method from a 1-mL culture grown overnight at 37☌ in lysogeny broth. ĭays From Onset of Symptoms to Date of Blood Culture Although NTS are usually associated with gastroenteritis in healthy people, individuals who are very young (1%), as estimated by CheckM version 1.0.11. Nontyphoidal salmonellae (NTS) are one of the leading causes of acute bacterial gastroenteritis worldwide. Bacteremia cluster, Salmonella enterica subsp enterica serovar Oranienburg
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