Multidrug‐resistant Pseudomonas aeruginosa is one of the main opportunistic pathogens causing severe infection. One of the mechanisms involved in the resistance to imipenem in clinical isolates is the loss of the OprD porin. Changes like substitutions, deletions, insertions, or mutations in the oprD gene can modify the conformation of OprD porin or inhibit its presence and generate resistance to carbapenems. The aim of this work was to obtain anti‐OprD polyclonal antibodies and to determine by both immunofluorescence microscopy (IFI) and Western blot assays, the presence of the OprD porin in resistant‐carbapenem P. aeruginosa strains with different changes in the oprD gene. Changes in the gene oprD were identified in clinical isolates of P. aeruginosa. When proteins were translated, several polymorphisms were found; however, these did not affect the presence of OprD porin (PCM25, PCM36, and PCM78). Also it was detected an insertion sequence ISPa1328 (PCM52) and a premature stop codon (PCM91), which inhibited the presence of the OprD porin. This study shows how changes in the oprD gene of P. aeruginosa clinical isolates affect the presence of the OprD porin detected by Western blot and indirect immunofluorescence assays using specific polyclonal anti‐OprD antibodies generated in this work.

中文翻译：

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OprD孔蛋白在铜绿假单胞菌临床分离株中的位置

多重耐药铜绿假单胞菌是引起严重感染的主要机会病原体之一。临床分离株对亚胺培南耐药的机制之一是OprD孔蛋白的损失。诸如oprD基因中的取代，缺失，插入或突变之类的变化可以修饰OprD孔蛋白的构象或抑制其存在并产生对碳青霉烯的抗性。这项工作的目的是获得抗OprD多克隆抗体，并通过免疫荧光显微镜（IFI）和Western印迹测定法确定在oprD基因中具有不同变化的抗性卡巴培南铜绿假单胞菌菌株中OprD孔蛋白的存在。基因oprD的变化在铜绿假单胞菌的临床分离物中鉴定到。翻译蛋白质时，发现了几种多态性。但是，这些不会影响OprD孔蛋白（PCM25，PCM36和PCM78）的存在。还检测到插入序列IS Pa1328（PCM52）和过早的终止密码子（PCM91），这抑制了OprD孔蛋白的存在。这项研究表明铜绿假单胞菌临床分离株的oprD基因的变化如何通过蛋白质印迹和间接免疫荧光分析检测到的OprD孔蛋白的存在，该蛋白使用这项工作中产生的特异性多克隆抗OprD抗体进行检测。