Flores-Oropeza Marco A, Ochoa Sara A, Cruz-Córdova Ariadnna, Chavez-Tepecano Rolando, Martínez-Peñafiel Eva, Rembao-Bojórquez Daniel, Zavala-Vega Sergio, Hernández-Castro Rigoberto, Flores-Encarnacion Marcos, Arellano-Galindo José, Vélez Daniel, Xicohtencatl-Cortes Juan
Posgrado en Ciencias Biomédicas, Instituto de Fisiología Celular, Universidad Nacional Autónoma de México, Mexico City, Mexico.
Laboratorio de Investigación en Bacteriología Intestinal, Unidad de Enfermedades Infecciosas, Hospital Infantil de México Federico Gómez, Mexico City, Mexico.
Front Microbiol. 2024 Jan 24;14:1340427. doi: 10.3389/fmicb.2023.1340427. eCollection 2023.
Recurrent urinary tract infections (RUTIs) caused by uropathogenic are costly public health problems impacting patients' quality of life.
In this work, a comparative genomics analysis of three clinical RUTI strains isolated from bladder biopsy specimens was performed.
One hundred seventy-two whole genomes of urinary tract strains were selected from the NCBI database. The search for virulence factors, fitness genes, regions of interest, and genetic elements associated with resistance was manually carried out. The phenotypic characterization of antibiotic resistance, haemolysis, motility, and biofilm formation was performed. Moreover, adherence and invasion assays with human bladder HTB-5 cells, and transmission electron microscopy (TEM) were performed.
The UTI-1_774U and UTI-3_455U/ST1193 strains were associated with the extraintestinal pathotypes, and the UTI-2_245U/ST295 strain was associated with the intestinal pathotype, according to a phylogenetic analysis of 172 urinary strains. The three RUTI strains were of clinical, epidemiological, and zoonotic relevance. Several resistance genes were found within the plasmids of these strains, and a multidrug resistance phenotype was revealed. Other virulence genes associated with CFT073 were not identified in the three RUTI strains (genes for type 1 and P fimbriae, haemolysin and toxin). Quantitative adherence analysis showed that UTI-1_774U was significantly ( < 0.0001) more adherent to human bladder HTB-5 cells. Quantitative invasion analysis showed that UTI-2_245U was significantly more invasive than the control strains. No haemolysis or biofilm activity was detected in the three RUTI strains. The TEM micrographs showed the presence of short and thin fimbriae only in the UTI-2_245U strain.
The high variability and genetic diversity of the RUTI strains indicate that are a mosaic of virulence, resistance, and fitness genes that could promote recurrence in susceptible patients.
由尿路致病性细菌引起的复发性尿路感染（RUTIs）是代价高昂的公共卫生问题，影响患者的生活质量。
在本研究中，对从膀胱活检标本中分离出的三株临床RUTI菌株进行了比较基因组学分析。
从NCBI数据库中选择了172株尿路菌株的全基因组。手动搜索毒力因子、适应性基因、感兴趣区域以及与耐药性相关的遗传元件。进行了抗生素耐药性、溶血、运动性和生物膜形成的表型特征分析。此外，还进行了与人膀胱HTB - 5细胞的黏附与侵袭试验以及透射电子显微镜（TEM）分析。
根据对172株尿路菌株的系统发育分析，UTI - 1_774U和UTI - 3_455U/ST1193菌株与肠外致病型相关，而UTI - 2_245U/ST295菌株与肠道致病型相关。这三株RUTI菌株具有临床、流行病学和人畜共患病学相关性。在这些菌株的质粒中发现了几个耐药基因，并揭示了多重耐药表型。在这三株RUTI菌株中未鉴定出其他与CFT073相关的毒力基因（1型菌毛和P菌毛、溶血素和毒素的基因）。定量黏附分析表明，UTI - 1_774U对人膀胱HTB - 5细胞的黏附显著更强（<0.0001）。定量侵袭分析表明，UTI - 2_245U的侵袭性明显高于对照菌株。在这三株RUTI菌株中未检测到溶血或生物膜活性。TEM显微照片显示仅在UTI - 2_245U菌株中存在短而细的菌毛。
RUTI菌株的高度变异性和遗传多样性表明，它们是毒力、耐药性和适应性基因的镶嵌体，可能促使易感患者复发。
