MOLECULAR CHARACTERIZATION BY PCR-RESTRICTION FRAGMENT LENGTH POLYMORPHISM OF TEM BETA-LACTAMASES
MOLECULAR CHARACTERIZATION BY PCR-RESTRICTION FRAGMENT LENGTH POLYMORPHISM OF TEM BETA-LACTAMASES
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DOI：
10.1016/0378-1097(95)00405-8
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发表时间：
1995-12-15
影响因子：
2.1
通讯作者：
PHILIPPON, A
中科院分区：
文献类型：
--
作者：
ARLET, G;BRAMI, G;PHILIPPON, A
为快速鉴定TEM型超广谱β-内酰胺酶，建立了一种快速简便的聚合酶链式反应-限制性片段长度多态性方法。用10种参考的TEM型超广谱β-内酰胺酶对该方法进行了验证。然后对之前仅通过生化分析报告的TEM-20和TEM-21中涉及的突变进行了表征。TEM-20与TEM-19的不同之处在于第925位有一个沉默的突变(A代表G)，而TEM-21与TEM-3和TEM-14的不同之处在于一个未报道的660位的单一突变(G代表A)，涉及到氨基酸取代，153位的组氨酸。此外，还描述了一种新型的对头孢他啶低耐药的超广谱β-内酰胺酶(TEM29)。TEM-29来源于TEM-1，氨基酸取代后为His-164。最后，结合聚合酶链式反应-限制性片段长度多态性和质粒分析，我们可以调查三家医院临床分离的多重耐药肺炎克雷伯菌所致的医院暴发。
To rapidly characterise TEM-derived extended-spectrum beta-lactamases a fast and easy method using polymerase chain reaction-restriction fragment length polymorphism was developed. This method was validated with ten reference TEM-type extended-spectrum beta-lactamases. The mutations involved in TEM-20 and TEM-21, which were previously reported only with biochemical analysis, were then characterised. TEM-20 differed from TEM-19 by a silent mutation at position 925 (A for G), and TEM-21 differed from TEM-3 and TEM-14 by a single mutation (G for A) in an unreported position 660, involving an amino acid substitution, arginine for histidine, at position 153. Moreover, a new extended-spectrum beta-lactamase conferring low resistance to ceftazidime (TEM-29), was described. TEM-29 derived from TEM-1, with an amino acid substitution, his-164. Finally, the combination of polymerase chain reaction-restriction fragment length polymorphism and plasmid analysis allowed us to investigate nosocomial outbreaks due to clinical isolates of multi-resistant Klebsiella pneumoniae in three hospitals.