Wang Haiping, Wu Dandan, Di Lingfang, Zhu Feiteng, Wang Zhengan, Sun Lu, Chen Yiyi, Jiang Shengnan, Zhuang Hemu, Chen Mengzhen, Ji Shujuan, Chen Yan
Department of Infectious Diseases, Sir Run Run Shaw Hospital, Zhejiang University School of Medicine, Hangzhou, China.
Key Laboratory of Microbial Technology and Bioinformatics of Zhejiang Province, Hangzhou, China.
Front Microbiol. 2022 Feb 28;13:814062. doi: 10.3389/fmicb.2022.814062. eCollection 2022.
To determine the genetic structure of -positive Tn1546-like mobile elements in methicillin-resistant (MRSA) from mainland China.
A total of 271 erythromycin-resistant MRSA isolates were isolated from Sir Run Run Shaw Hospital (SRRSH) from 2013 to 2015. Whole-genome sequencing was performed for the -positive strains, and the genetic environment of the genes was analyzed. Southern hybridization analysis and transformation tests were performed to confirm the location of the gene.
A total of 64 isolates (64/271, 23.6%) were -positive strains, with 62 strains (62/64, 96.9%) belonging to the CC59 clone. The other two strains, SR130 and SR231, belonging to CC5-ST965, both harbored 14,567 bp -positive Tn1546-like elements and displayed multidrug-resistant profiles. PFGE followed by Southern blot demonstrated that the genes were located on the plasmids of both SR130 and SR231, while two copies of ermB were located on the chromosome of SR231. Further sequencing demonstrated that SR231 carried one Tn1546- elements in the plasmid and two identical copies integrated on the chromosome, which had 99.99% identity to the element in the plasmid of SR130. The Tn1546- elements were highly similar (100% coverage, >99.9% identity) to the element Tn6636 reported in a previous study from Taiwan. The plasmids (pSR130 and pSR231) harboring -positive Tn1546-like elements were also identical to the mosaic plasmid pNTUH_5066148. However, conjugation of -carrying plasmids of SR130 and SR231 were failed after triple repeats.
Multiple copies of -positive Tn1546-like mobile elements were found in CC5-ST965 MRSA from mainland China, showing the wide dissemination of these -originated -positive Tn1546-like elements. Molecular epidemiological study of Tn1546-like elements is essential to avoid the spreading of resistant determinants.
确定中国大陆耐甲氧西林金黄色葡萄球菌(MRSA)中阳性Tn1546样移动元件的遗传结构。
2013年至2015年从邵逸夫医院(SRRSH)分离出271株耐红霉素MRSA菌株。对阳性菌株进行全基因组测序,并分析erm基因的遗传环境。进行Southern杂交分析和转化试验以确认erm基因的位置。
共有64株(64/271,23.6%)为阳性菌株,其中62株(62/64,96.9%)属于CC59克隆。另外两株,SR130和SR231,属于CC5-ST965,均携带14567 bp的阳性Tn1546样元件,并表现出多重耐药谱。脉冲场凝胶电泳(PFGE)结合Southern印迹显示,erm基因位于SR130和SR231的质粒上,而两个ermB拷贝位于SR231的染色体上。进一步测序表明,SR231在质粒中携带一个Tn1546-like元件,在染色体上整合了两个相同的拷贝,与SR130质粒中的元件具有99.99%的同一性。Tn1546-like元件与台湾先前一项研究中报道的Tn6636元件高度相似(100%覆盖,>99.9%同一性)。携带阳性Tn1546样元件的质粒(pSR130和pSR231)也与嵌合质粒pNTUH_5066148相同。然而,SR130和SR231携带erm的质粒在三次重复后接合失败。
在中国大陆CC5-ST965 MRSA中发现了多个拷贝的阳性Tn1546样移动元件,表明这些源自erm的阳性Tn1546样元件广泛传播。对Tn1546样元件进行分子流行病学研究对于避免耐药决定簇的传播至关重要。