Adiguzel Mehmet Cemal, Goulart Debora Brito, Wu Zuowei, Pang Jinji, Cengiz Seyda, Zhang Qijing, Sahin Orhan
Department of Microbiology, College of Veterinary Medicine, Ataturk University, Erzurum 25240, Turkey.
Department of Veterinary Microbiology and Preventive Medicine, College of Veterinary Medicine, Iowa State University, Ames, IA 50011, USA.
Pathogens. 2021 Mar 16;10(3):345. doi: 10.3390/pathogens10030345.
To aid development of phage therapy against , we investigated the distribution of the clustered regularly interspaced short palindromic repeats (CRISPR) systems in fluoroquinolone (FQ)-resistant . A total of 100 FQ-resistant strains from different sources were analyzed by PCR and DNA sequencing to determine resistance-conferring mutation in the gene and the presence of various CRISPR systems. All but one isolate harbored 1-5 point mutations in and the most common mutation was the Thr86Ile change. Ninety-five isolates were positive with the CRISPR PCR, and spacer sequences were found in 86 of them. Among the 292 spacer sequences identified in this study, 204 shared 93-100% nucleotide homology to phage D10, 44 showed 100% homology to phage CP39, and 3 had 100% homology with phage CJIE4-5. The remaining 41 spacer sequences did not match with any phages in the database. Based on the results, it was inferred that the FQ-resistant isolates analyzed in this study were potentially resistant to phages D10, CP39, and CJIE4-5 as well as some unidentified phages. These phages should be excluded from cocktails of phages that may be utilized to treat FQ-resistant .
为了助力噬菌体疗法针对[具体病原体]的开发,我们研究了耐氟喹诺酮(FQ)[具体病原体]中规律成簇间隔短回文重复序列(CRISPR)系统的分布情况。通过聚合酶链式反应(PCR)和DNA测序对总共100株来自不同来源的耐FQ[具体病原体]菌株进行分析,以确定[具体基因]中赋予耐药性的突变以及各种CRISPR系统的存在情况。除了一株分离株外,所有分离株在[具体基因]中都存在1至5个点突变,最常见的突变是Thr86Ile改变。95株分离株的CRISPR PCR检测呈阳性,其中86株发现有间隔序列。在本研究鉴定出的292个间隔序列中,204个与噬菌体D10具有93%至100%的核苷酸同源性,44个与噬菌体CP39具有100%的同源性,3个与噬菌体CJIE4 - 5具有100%的同源性。其余41个间隔序列与数据库中的任何噬菌体均不匹配。基于这些结果,推测本研究中分析的耐FQ[具体病原体]分离株可能对噬菌体D10、CP39和CJIE4 - 5以及一些未鉴定的噬菌体具有抗性。这些噬菌体应从可能用于治疗耐FQ[具体病原体]的噬菌体混合物中排除。