Maszewska Agnieszka, Wójcik Ewelina, Ciurzyńska Aneta, Wojtasik Arkadiusz, Piątkowska Iwona, Dastych Jarosław, Różalski Antoni
Department of Immunobiology of Bacteria, Faculty of Biology and Environmental Protection, University of Lodz, Łódź, Poland.
Proteon Pharmaceuticals SA, Łódź, Poland.
Acta Biochim Pol. 2016;63(2):303-10. doi: 10.18388/abp.2015_1114. Epub 2016 Jan 7.
Urinary tract infections (UTIs) caused by P. mirabilis are difficult to cure because of the increasing antimicrobial resistance of these bacteria. Phage therapy is proposed as an alternative infection treatment. The aim of this study was to isolate and differentiate uropathogenic P. mirabilis strain specific polyvalent bacteriophages producing polysaccharide depolymerases (PDs). 51 specific phages were obtained. The plaques of 29 bacteriophages were surrounded by halos, which indicated that they produced PDs. The host range analysis showed that, except phages 58B and 58C, the phage host range profiles differed from each other. Phages 35 and 45 infected all P. mirabilis strains tested. Another 10 phages lysed more than 90% of isolates. Among these phages, 65A, 70, 66 and 66A caused a complete lysis of the bacterial lawn formed by 62% to 78% of strains. Additionally, phages 39A and 70 probably produced PDs. The phages' DNA restriction fragment length polymorphism (RFLP) analysis demonstrated that genomes of 51 isolated phages represented 34 different restriction profiles. DNA of phage 58A seemed to be resistant to selected EcoRV endonuclease. The 33 RFLP-EcoRV profiles showed a Dice similarity index of 38.8%. 22 RFLP patterns were obtained from single phage isolates. The remaining 12 restriction profiles consisted of 2 to 4 viruses. The results obtained from phage characterization based on the pattern of phage host range in combination with the RFLP method enabled effective differentiation of the studied phages and selection of PD producing polyvalent phages for further study.
奇异变形杆菌引起的尿路感染(UTIs)由于这些细菌日益增强的抗菌耐药性而难以治愈。噬菌体疗法被提议作为一种替代性的感染治疗方法。本研究的目的是分离和鉴别产生多糖解聚酶(PDs)的尿路致病性奇异变形杆菌菌株特异性多价噬菌体。获得了51种特异性噬菌体。29种噬菌体的噬菌斑周围有晕圈,这表明它们产生了PDs。宿主范围分析表明,除了噬菌体58B和58C外,噬菌体的宿主范围图谱彼此不同。噬菌体35和45感染了所有测试的奇异变形杆菌菌株。另外10种噬菌体裂解了90%以上的分离株。在这些噬菌体中,65A、70、66和66A导致由62%至78%的菌株形成的菌苔完全裂解。此外,噬菌体39A和70可能产生了PDs。噬菌体的DNA限制性片段长度多态性(RFLP)分析表明,51种分离噬菌体的基因组代表了34种不同的限制性图谱。噬菌体58A的DNA似乎对选定的EcoRV核酸内切酶具有抗性。33种RFLP-EcoRV图谱的Dice相似性指数为38.8%。从单个噬菌体分离物中获得了22种RFLP模式。其余12种限制性图谱由2至4种病毒组成。基于噬菌体宿主范围模式结合RFLP方法进行噬菌体鉴定所获得的结果,能够有效地鉴别所研究的噬菌体,并选择产生PDs的多价噬菌体进行进一步研究。