来自克罗地亚一家三级医院的耐碳青霉烯类和碳青霉烯类敏感临床分离株中的CRISPR-Cas动态变化

CRISPR-Cas Dynamics in Carbapenem-Resistant and Carbapenem-Susceptible Clinical Isolates from a Croatian Tertiary Hospital.

作者信息

Jurić Ivana, Jelić Marko, Markanović Manda, Kanižaj Lucija, Bošnjak Zrinka, Budimir Ana, Kuliš Tomislav, Tambić-Andrašević Arjana, Ivančić-Baće Ivana, Mareković Ivana

机构信息

Clinical Microbiology, Infection Prevention and Control Department, University Hospital Centre Zagreb, Kišpatićeva 12, 10000 Zagreb, Croatia.

Department of Clinical Microbiology, University Hospital for Infectious Diseases "Fran Mihaljević", Mirogojska 8, 10000 Zagreb, Croatia.

出版信息

Pathogens. 2025 Jun 19;14(6):604. doi: 10.3390/pathogens14060604.

DOI:10.3390/pathogens14060604

PMID:40559612

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC12195874/

Abstract

(1) Background: CRISPR-Cas systems provide adaptive immunity against mobile genetic elements (MGEs) carrying antimicrobial resistance (AMR) genes. Carbapenem-resistant (CR) is a major public health concern, and the role of CRISPR-Cas in its resistance is understudied. This study explored CRISPR-Cas associations with multidrug resistance in clinical . (2) Methods: 400 isolates (200 CR and 200 carbapenem susceptible (CS)) were analyzed. Carbapenemase genes (, , ), , , and CRISPR1-3 loci were identified by PCR, while only CRISPR loci were sequenced. Genetic relatedness was assessed via PFGE, MLST, and spacer analysis. Statistical analysis utilized chi-squared and Fisher's exact tests. (3) Results: CRISPR-Cas was present in 15.8% of isolates, mainly subtypes I-E and I-E* (93.3%), with CRISPR3 loci showing the greatest spacer diversity. Clonal complexes ST14/15/101 (CR) and ST35 (CS) were identified. was linked to CRISPR-Cas-negative strains, while and were more frequent in CRISPR-Cas-positive strains ( < 0.0001). Imipenem/relebactam resistance was higher in CRISPR-Cas-negative isolates. (4) Conclusions: CRISPR-Cas systems correlate with specific carbapenemase profiles, suggesting pressure against acquisition. The coexistence of I-E and I-E* subtypes highlight synergies in targeting MGEs. CRISPR loci could be tools for subtyping organisms following MLST.

摘要

(1) 背景：CRISPR-Cas系统可对携带抗菌药物耐药性（AMR）基因的移动遗传元件（MGEs）提供适应性免疫。耐碳青霉烯类（CR）是一个主要的公共卫生问题，而CRISPR-Cas在其耐药性中的作用尚未得到充分研究。本研究探讨了CRISPR-Cas与临床多重耐药性的关联。(2) 方法：分析了400株分离株（200株CR和200株碳青霉烯类敏感株（CS））。通过PCR鉴定碳青霉烯酶基因（blaKPC、blaNDM、blaOXA-48）、blaVIM、blaIMP和CRISPR1-3位点，而仅对CRISPR位点进行测序。通过脉冲场凝胶电泳（PFGE）、多位点序列分型（MLST）和间隔区分析评估遗传相关性。统计分析采用卡方检验和Fisher精确检验。(3) 结果：15.8%的分离株中存在CRISPR-Cas，主要亚型为I-E和I-E*（93.3%），CRISPR3位点的间隔区多样性最大。鉴定出克隆复合体ST14/15/101（CR）和ST35（CS）。blaKPC与CRISPR-Cas阴性菌株相关，而blaNDM和blaOXA-48在CRISPR-Cas阳性菌株中更常见（P<0.0001）。CRISPR-Cas阴性分离株对亚胺培南/瑞来巴坦的耐药性更高。(4) 结论：CRISPR-Cas系统与特定的碳青霉烯酶谱相关，提示对blaKPC获得存在压力。I-E和I-E*亚型的共存突出了在靶向MGEs方面的协同作用。CRISPR位点可作为MLST之后对生物体进行亚型分型的工具。