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：
10.1111/j.1574-6968.1995.tb07938.x
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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
为了快速表征 TEM 衍生的广谱 β-内酰胺酶，开发了一种使用聚合酶链反应-限制性片段长度多态性的快速简便方法。该方法已用 10 种参考 TEM 型广谱 β-内酰胺酶进行了验证。然后对涉及 TEM-20 和 TEM-21 的突变进行了表征，这些突变以前仅通过生化分析进行报告。 TEM-20 与 TEM-19 的不同之处在于第 925 位处的沉默突变（A 为 G），而 TEM-21 与 TEM-3 和 TEM-14 的不同之处在于未报告的第 660 位处的单一突变（G 为 A），涉及第 153 位的氨基酸替换，精氨酸替换为组氨酸。此外，一种新的广谱 β-内酰胺酶赋予头孢他啶低耐药性(TEM-29)，进行了描述。 TEM-29 源自 TEM-1，带有氨基酸取代 his-164。最后，聚合酶链反应-限制性片段长度多态性和质粒分析相结合，使我们能够调查三家医院因多重耐药肺炎克雷伯菌临床分离株而导致的医院暴发。
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.