Tsakris Athanassios, Ikonomidis Alexandros, Spanakis Nicholas, Poulou Aggeliki, Pournaras Spyros
Department of Microbiology, Medical School, University of Athens, Athens, Greece.
J Antimicrob Chemother. 2007 Apr;59(4):739-41. doi: 10.1093/jac/dkm020. Epub 2007 Mar 6.
A carbapenem-resistant Morganella morganii clinical isolate that was phenotypically metallo-beta-lactamase (MBL)-positive was recovered from a Greek patient. The aim of the study was to analyse the structure of the integron containing the MBL gene.
MICs were determined by the broth microdilution method. PCR assays and nucleotide sequencing were used for identification of bla gene types and mapping of the integron carrying the MBL gene. The location of the MBL allele was investigated by mating experiments and plasmid analysis as well as by Southern blotting of the plasmid extract and gene-specific hybridization with a bla(VIM-1) probe.
The strain contained In3Mor, a novel class 1 integron carrying a carbapenemase gene (bla(VIM-1)) associated with a trimethoprim (dfrA1), a streptothricin (sat1) and two aminoglycoside resistance genes (aacA7 and aadA1). Conjugation experiments failed to detect a transferable MBL determinant and plasmid DNA was not visualized. The chromosomal location of bla(VIM-1) was confirmed after hybridization of the chromosomal band with the bla(VIM-1) probe.
Production of a VIM-type MBL in a M. morganii clinical isolate is documented in this study for the first time. Also, the dfrA1-sat1-aadA1 array which is typically described in the variable region of class 2 integrons consistent with that on Tn7 transposons, is originally detected herein in a class 1 integron.
从一名希腊患者身上分离出一株耐碳青霉烯类摩根氏摩根菌临床分离株,其表型为金属β-内酰胺酶(MBL)阳性。本研究的目的是分析携带MBL基因的整合子结构。
采用肉汤微量稀释法测定最低抑菌浓度(MIC)。采用聚合酶链反应(PCR)检测和核苷酸测序鉴定bla基因类型,并对携带MBL基因的整合子进行定位。通过接合试验、质粒分析以及对质粒提取物进行Southern印迹分析并用bla(VIM-1)探针进行基因特异性杂交,研究MBL等位基因的位置。
该菌株含有In3Mor,这是一种新型1类整合子,携带与甲氧苄啶(dfrA1)、链丝菌素(sat1)以及两个氨基糖苷类耐药基因(aacA7和aadA1)相关的碳青霉烯酶基因(bla(VIM-1))。接合试验未能检测到可转移的MBL决定簇,也未观察到质粒DNA。用bla(VIM-1)探针与染色体条带杂交后,证实了bla(VIM-1)的染色体定位。
本研究首次报道了摩根氏摩根菌临床分离株产生VIM型MBL。此外,通常在2类整合子可变区中描述的与Tn7转座子上一致的dfrA1-sat1-aadA1阵列,在本研究中首次在1类整合子中检测到。