Research Institute for Systems Biology and Medicine, 117246 Moscow, Russia.
Research Institute for Systems Biology and Medicine, 117246 Moscow, Russia.
J Microbiol Methods. 2024 Dec;227:107054. doi: 10.1016/j.mimet.2024.107054. Epub 2024 Oct 11.
The transfer of biocide and antibiotic resistance genes by mobile genetic elements is the most common mechanism for rapidly acquiring and spreading resistance among bacteria. The qacEΔ1 gene confers the resistance to quaternary ammonium compounds (QACs). It has also been considered a genetic marker for the presence of class 1 integrons associated with multidrug-resistant (MDR) phenotypes in Gram-negative bacteria. In this study, a TaqMan real-time PCR assay was developed to detect the qacEΔ1 gene in Gram-negative bacteria. The assay has a detection limit of 80 copies of the qacEΔ1 gene per reaction. No false-positive or false-negative results have been observed. Simultaneous amplification and detection of the 16S rRNA gene is performed as an endogenous internal amplification control (IAC). The TaqMan real-time PCR assay developed is a rapid, sensitive, and specific method that could be used to monitor resistance to QACs, the spread of class 1 integrons, and the prediction of associated MDR phenotypes in Gram-negative bacteria.
移动遗传元件介导的杀菌药物和抗生素耐药基因的转移是细菌快速获得和传播耐药性的最常见机制。qacEΔ1 基因赋予了对季铵化合物 (QACs) 的耐药性。它也被认为是与革兰氏阴性菌中多药耐药 (MDR) 表型相关的 1 类整合子存在的遗传标记。在本研究中，开发了一种 TaqMan 实时 PCR 检测方法来检测革兰氏阴性菌中的 qacEΔ1 基因。该检测方法的检测限为每个反应 80 个 qacEΔ1 基因拷贝。没有观察到假阳性或假阴性结果。16S rRNA 基因的同时扩增和检测作为内源性内部扩增对照 (IAC)。开发的 TaqMan 实时 PCR 检测方法是一种快速、敏感和特异的方法，可用于监测革兰氏阴性菌对 QACs 的耐药性、1 类整合子的传播以及相关 MDR 表型的预测。
