Department of Microbiology, School of Basic Medical Sciences, Hubei University of Medicine, Hubei, Shiyan 442000, China; Department of Clinical Laboratory, Affiliated dongfeng Hospital, Hubei University of Medicine, Hubei, Shiyan 442000, China; Hubei Key Laboratory of Wudang Local Chinese Medicine Research, Hubei, Shiyan 442000, China.
Department of Microbiology, School of Basic Medical Sciences, Hubei University of Medicine, Hubei, Shiyan 442000, China.
J Glob Antimicrob Resist. 2020 Sep;22:835-841. doi: 10.1016/j.jgar.2020.06.022. Epub 2020 Jul 8.
To identify the general features of acquisition of drug-resistance genes in two multi-drug resistant Enterobacteriaceae strains isolated from a single patient in China.
The whole-plasmid was sequenced by Illumina Hiseq 4000 and Pacbio RSII procedures. The plasmid conjugation transfer experiment were performed by the mating-out assay. Drug-resistance genes were amplified by PCR assay.
We identified two New Delhi metallo-β-lactamase type 1(NDM-1)-producing isolates, named Raoultella ornithinolytica B1645-1 and Enterobacter cloacae B1645-2, which shared the same sulfonamide-resistant dihydropteroate synthase sul2 gene and aminoglycoside O-phosphotransferase aph(3'')-Ib gene. A novel antimicrobial resistance plasmid pCYNDM01 was first discovered from the multi-drug resistant R. ornithinolytica B1645-1. Interestingly, plasmid pCYNDM01 carried a Gifsy-2 prophage gene. The bla gene was located on a novel complex class 1 integron with a structure of sul1-qacEΔ1-ΔISAba125-bla-bla-trpC-ISCR1-catb8-aacA4-IS1-IS6100-dfrA14-intI1. The carrying the bla gene plasmid pCYNDM01 was transferred to the E. cloacae B1645-2 recipient strain. This 149.44 kb plasmid pCYNDM01 belonged to the IncFII type.
A novel antimicrobial resistance plasmid pCYNDM01 was first recovered from a multi-drug resistance R. ornithinolytica B1645-1 isolated from China. The novel complex sul1-type class 1 integron might play an essential role in the mobilization of the bla gene among different enterobacterial species. The occurrence of plasmid pCYNDM01 transfer from R. ornithinolytica to E. cloacae in vitro by conjugation showed that plasmid pCYNDM01 was a self-conjugative plasmid and might cause dissemination of drug-resistance genes within different enterobacterial species from a single patient in vivo by conjugation. The novel variant F-like T4SS of plasmid pCYNDM01 might be as a tool of R. ornithinolytica B1645-1 for resistance genes transfer. The emergence of the two NDM-1-producing Enterobacteriaceae strains should be attracted China attentions and required to prevent its future prevalence.
鉴定从中国的单一患者中分离出的两株多药耐药肠杆菌科菌株中耐药基因的获得的一般特征。
采用 Illumina Hiseq 4000 和 Pacbio RSII 程序对全质粒进行测序。通过交配实验进行质粒接合转移实验。通过 PCR 实验扩增耐药基因。
我们鉴定了两株新德里金属-β-内酰胺酶 1 型（NDM-1）产生的分离株，命名为鸟氨酸罗伊氏菌 B1645-1 和阴沟肠杆菌 B1645-2，它们共享相同的磺胺类耐药二氢喋呤合成酶 sul2 基因和氨基糖苷磷酸转移酶 aph（3'）-Ib 基因。首次从多药耐药的 R. ornithinolytica B1645-1 中发现了一种新型的抗菌药物耐药质粒 pCYNDM01。有趣的是，质粒 pCYNDM01 携带了 Gifsy-2 噬菌体基因。bla 基因位于一个新型的复杂 1 类整合子上，其结构为 sul1-qacEΔ1-ΔISAba125-bla-bla-trpC-ISCR1-catb8-aacA4-IS1-IS6100-dfrA14-intI1。携带 bla 基因的质粒 pCYNDM01 被转移到阴沟肠杆菌 B1645-2 受体菌株中。这个 149.44kb 的质粒 pCYNDM01 属于 IncFII 型。
首次从中国多药耐药的 R. ornithinolytica B1645-1 中回收了一种新型的抗菌药物耐药质粒 pCYNDM01。新型复杂的 sul1 型 1 类整合子可能在不同肠杆菌种之间 bla 基因的转移中起重要作用。通过接合，质粒 pCYNDM01 从鸟氨酸罗伊氏菌转移到阴沟肠杆菌的体外转移表明，质粒 pCYNDM01 是一种自我接合性质粒，可能通过接合在体内从单一患者的不同肠杆菌种中传播耐药基因。质粒 pCYNDM01 的新型 F 样 T4SS 可能是 R. ornithinolytica B1645-1 用于耐药基因转移的工具。两株产 NDM-1 的肠杆菌科菌株的出现应引起中国的关注，并需要防止其未来流行。
