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BlaEC家族C类β-内酰胺酶分析

Analysis of BlaEC family class C beta-lactamase.

作者信息

Schmidt Jiri, Zdarska Veronika, Kolar Milan, Mlynarcik Patrik

机构信息

Department of Biotechnology, Faculty of Science, Palacky University Olomouc, 17. listopadu 1192/12, 779 00 Olomouc, Czechia.

Department of Microbiology, Faculty of Medicine and Dentistry, Palacky University Olomouc, Hnevotinska 3, 779 00 Olomouc, Czechia.

出版信息

FEMS Microbiol Lett. 2023 Jan 17;370. doi: 10.1093/femsle/fnad097.

Abstract

Recent years have witnessed an increased prevalence of intrinsic and acquired beta-lactamase-producing bacteria, severely limiting human and veterinary medicine therapeutic options. The present study aimed to design specific oligonucleotides for rapid PCR detection of the cephalosporinase-encoding gene blaEC (BlaEC family class C beta-lactamase). A total of three primers were designed to detect 2281 variants of the blaEC gene and two sets of primer pairs were also tested against DNA from 11 strains. The study indicates that the proposed primers should be able to detect 100% of all described blaEC genes in different bacterial strains and monitor their spread. After comparing the amino acid sequences, a phylogenetic tree was created based on the presence of conserved amino acids and homologous motifs. More than 24 760 mutations in BlaEC enzymes have been identified. The mutations involving 371 amino acid positions and these hotspots can change the structure and activity of the monitored enzymes. We predicted several BlaEC enzymes with a broadened substrate activity against higher-generation cephalosporins.

摘要

近年来,产内源性和获得性β-内酰胺酶细菌的患病率有所上升,严重限制了人类和兽医学的治疗选择。本研究旨在设计特异性寡核苷酸,用于快速PCR检测编码头孢菌素酶的基因blaEC(BlaEC家族C类β-内酰胺酶)。总共设计了三种引物来检测blaEC基因的2281个变体,还针对11株菌株的DNA测试了两组引物对。该研究表明,所提出的引物应该能够检测不同细菌菌株中所有已描述的blaEC基因的100%,并监测它们的传播。在比较氨基酸序列后,基于保守氨基酸和同源基序的存在构建了系统发育树。已鉴定出BlaEC酶中超过24760个突变。涉及371个氨基酸位置的这些突变和热点可改变所监测酶的结构和活性。我们预测了几种对更高代头孢菌素有更广泛底物活性的BlaEC酶。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dfbe/10563145/2597d3a9b6dc/fnad097fig1.jpg

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