De Lappe N, Doran G, O'Connor J, O'Hare C, Cormican M
National Salmonella Reference Laboratory, Galway, Republic of Ireland.
National University of Ireland Galway, Galway, Republic of Ireland.
J Med Microbiol. 2009 Jan;58(Pt 1):86-93. doi: 10.1099/jmm.0.000034-0.
The 16 Salmonella enterica serovar Enteritidis (S. Enteritidis) typing phages (SETPs) used in the Laboratory of Enteric Pathogens (Health Protection Agency, London, UK) phage-typing scheme have not previously been characterized in detail. We have examined the adsorption properties of the phages with respect to a number of S. enterica serovars and defined phage morphology with electron microscopy. PFGE was used to estimate overall genome size and banding patterns generated by electrophoresis following restriction endonuclease digestion of the genome with HindIII were compared. PCR amplification and sequencing of selected genes was performed. The 16 phages comprise three morphotypes, Podoviridae (SETP1, 8, 10, 14, 15 and 16), Siphoviridae (SETP3, 5, 7, 11, 12 and 13) and Myoviridae (SETP2, 4, 6 and 9). All Podoviridae and Siphoviridae, but not Myoviridae, adsorbed to the O12 lipopolysaccharide antigen of Salmonella serogroups B (4,12) and D(1) (9,12). The genome sizes for the Podoviridae and Siphoviridae (PFGE-A) were approximately 42 kb. The genome size for Myoviridae SETP2, 4 and 9 was 36.5 kb, and for myovirus SETP6 was 27 kb. HindIII digestion of phage DNA produced 9 distinct patterns of 8 to 11 bands. Relationships between phages based on digest patterns were consistent with those defined by morphology. The Podoviridae had homologues of several P22 genes while the Siphoviridae had homologues of several genes present in the sequenced siphovirus SETP3 (EF177456). This study represents an initial step in characterizing the molecular basis that underlies the widely used S. Enteritidis typing scheme.
用于肠道病原体实验室（英国伦敦卫生防护局）噬菌体分型方案的16种肠炎沙门氏菌肠炎血清型噬菌体（SETPs），此前尚未进行过详细的特性分析。我们研究了这些噬菌体对多种肠炎沙门氏菌血清型的吸附特性，并用电子显微镜确定了噬菌体形态。采用脉冲场凝胶电泳（PFGE）估计基因组总大小，并比较了用HindIII对基因组进行限制性内切酶消化后电泳产生的条带模式。对选定基因进行了PCR扩增和测序。这16种噬菌体包括三种形态类型：短尾噬菌体科（SETP1、8、10、14、15和16）、长尾噬菌体科（SETP3、5、7、11、12和13）和肌尾噬菌体科（SETP2、4、6和9）。所有短尾噬菌体科和长尾噬菌体科噬菌体，但不包括肌尾噬菌体科噬菌体，都能吸附到沙门氏菌B（4,12）和D（1）（9,12）血清群的O12脂多糖抗原上。短尾噬菌体科和长尾噬菌体科（PFGE-A）的基因组大小约为42 kb。肌尾噬菌体科SETP2、4和9的基因组大小为36.5 kb，肌尾噬菌体SETP6的基因组大小为27 kb。噬菌体DNA的HindIII消化产生了9种不同的条带模式，条带数为8至11条。基于消化模式的噬菌体之间的关系与形态学定义的关系一致。短尾噬菌体科有几个P22基因的同源物，而长尾噬菌体科有几个存在于已测序的长尾噬菌体SETP3（EF177456）中的基因的同源物。本研究是对广泛使用的肠炎沙门氏菌分型方案背后的分子基础进行特性分析的第一步。
