Mlynarcik Patrik, Chudobova Hana, Zdarska Veronika, Kolar Milan
Department of Microbiology, Faculty of Medicine and Dentistry, Palacky University Olomouc, Hnevotinska 3, 77515 Olomouc, Czech Republic.
Laboratory of Growth Regulators, Faculty of Science, Institute of Experimental Botany of the Czech Academy of Sciences, Palacky University, Šlechtitelů 27, 78371 Olomouc, Czech Republic.
Antibiotics (Basel). 2021 Jul 4;10(7):812. doi: 10.3390/antibiotics10070812.
The growing bacterial resistance to available β-lactam antibiotics is a very serious public health problem, especially due to the production of a wide range of β-lactamases. At present, clinically important bacteria are increasingly acquiring new elements of resistance to carbapenems and polymyxins, including extended-spectrum β-lactamases (ESBLs), carbapenemases and phosphoethanolamine transferases of the MCR type. These bacterial enzymes limit therapeutic options in human and veterinary medicine. It must be emphasized that there is a real risk of losing the ability to treat serious and life-threatening infections. The present study aimed to design specific oligonucleotides for rapid PCR detection of ESBL-encoding genes and in silico analysis of selected ESBL enzymes. A total of 58 primers were designed to detect 49 types of different ESBL genes. After comparing the amino acid sequences of ESBLs (CTX-M, SHV and TEM), phylogenetic trees were created based on the presence of conserved amino acids and homologous motifs. This study indicates that the proposed primers should be able to specifically detect more than 99.8% of all described ESBL enzymes. The results suggest that the in silico tested primers could be used for PCR to detect the presence of ESBL genes in various bacteria, as well as to monitor their spread.
细菌对现有β-内酰胺类抗生素的耐药性不断增强是一个非常严重的公共卫生问题,尤其是由于多种β-内酰胺酶的产生。目前,临床上重要的细菌越来越多地获得对碳青霉烯类和多粘菌素的新耐药机制,包括超广谱β-内酰胺酶(ESBLs)、碳青霉烯酶和MCR型磷酸乙醇胺转移酶。这些细菌酶限制了人类和兽医学中的治疗选择。必须强调的是,确实存在失去治疗严重和危及生命感染能力的风险。本研究旨在设计用于快速PCR检测ESBL编码基因的特异性寡核苷酸,并对选定的ESBL酶进行计算机分析。总共设计了58对引物来检测49种不同的ESBL基因。在比较ESBLs(CTX-M、SHV和TEM)的氨基酸序列后,基于保守氨基酸和同源基序的存在构建了系统发育树。本研究表明,所提出的引物应该能够特异性地检测所有已描述的ESBL酶中的99.8%以上。结果表明,经计算机测试的引物可用于PCR检测各种细菌中ESBL基因的存在,并监测其传播。