Karama Musafiri, Gyles Carlton L
Department of Pathobiology, Ontario Veterinary College, University of Guelph, Guelph, ON N1G 2W1, Canada.
Appl Environ Microbiol. 2008 Aug;74(16):5153-8. doi: 10.1128/AEM.00723-08. Epub 2008 Jun 13.
The objectives of this study were to induce and characterize verotoxin-encoding phages from a collection of 91 verotoxin-producing Escherichia coli (VTEC) O103:H2 strains of human and bovine origins. All the strains carried the vt1 gene, and two carried the vt2 gene as well. The phages were induced by UV irradiation and characterized by DNA restriction fragment length polymorphism (RFLP), genome size, morphology, and Q and P genes, characteristic of lambdoid phages. A total of 32 vt-positive phages were induced and isolated from 31 VTEC O103:H2 strains. Thirty phages were vt1 positive, and two were vt2 positive. Ten of the 30 vt1-positive phages (33.3%) were from cattle strains, and 20 (66.6%) were from human strains. The two vt2-positive phages were from human strains. Phages belonged to 21 RFLP profiles, of which 17 were single-phage profiles and 4 were multiple-phage profiles. The estimated genome size of the phages ranged from 34 to 84 kb. Two phages that were examined by electron microscopy possessed hexagonal heads with long tails, and one had an elongated head with a long tail. The Q and P genes were amplified in all 32 phages, and the Q-stxA(1) gene region yielded an amplicon in 19 phages (59.3%). It is concluded that the VTEC O103:H2 strains of human origin were more readily inducible than those of bovine origin and that the genotypic profiles of verotoxin-encoding phages were highly diverse, as revealed by their RFLP profiles.
本研究的目的是从91株来自人和牛源的产志贺毒素大肠杆菌(VTEC)O103:H2菌株中诱导并鉴定编码志贺毒素的噬菌体。所有菌株都携带vt1基因,其中两株还携带vt2基因。通过紫外线照射诱导噬菌体,并通过DNA限制性片段长度多态性(RFLP)、基因组大小、形态以及λ样噬菌体特有的Q和P基因进行鉴定。共从31株VTEC O103:H2菌株中诱导并分离出32株vt阳性噬菌体。30株噬菌体为vt1阳性,2株为vt2阳性。30株vt1阳性噬菌体中,10株(33.3%)来自牛源菌株,20株(66.6%)来自人源菌株。两株vt2阳性噬菌体均来自人源菌株。噬菌体属于21种RFLP图谱类型,其中17种为单噬菌体图谱类型,4种为多噬菌体图谱类型。噬菌体的估计基因组大小在34至84 kb之间。通过电子显微镜检查的两株噬菌体具有六边形头部和长尾,一株具有细长头部和长尾。所有32株噬菌体中均扩增出Q和P基因,19株噬菌体(59.3%)的Q-stxA(1)基因区域产生扩增产物。得出的结论是,人源VTEC O103:H2菌株比牛源菌株更容易被诱导,并且编码志贺毒素的噬菌体的基因型图谱高度多样,这从它们的RFLP图谱中可以看出。