Classification of OprD sequence and correlation with antimicrobial activity of carbapenem agents in Pseudomonas aeruginosa clinical isolates collected in Japan
Classification of OprD sequence and correlation with antimicrobial activity of carbapenem agents in Pseudomonas aeruginosa clinical isolates collected in Japan
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DOI：
10.1111/j.1348-0421.2009.00137.x
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发表时间：
2009-07-01
影响因子：
2.6
通讯作者：
Gotoh, Naomasa
中科院分区：
文献类型：
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作者：
Sanbongi, Yumiko;Shimizu, Atsuyuki;Gotoh, Naomasa
研究了1998年至2001年间在日本收集的总共99株金属β-内酰胺酶阴性铜绿假单胞菌临床分离株对碳青霉烯类药物的敏感性以及相应的oprD基因突变。根据改变的模式，每个菌株的 OprD 序列分为两大类。 80 个菌株（80.8%）是所谓的“全长型”，其 OprD 蛋白被完全编码。其余 19 个菌株（19.2%）是所谓的“缺陷型”，它们具有可能导致 OprD 孔蛋白构象变化的缺失或重大改变。与“全长型”菌株相比，“缺陷型”菌株的变化导致亚胺培南、美罗培南和比阿培南的几何平均 MIC 分别增加了 15 倍、17 倍和 23 倍。 “全长型”菌株进一步分为六种碳青霉烯类敏感类型，四种碳青霉烯类抗性亚型除外，在 D43、G183、R154、G314、G316 处有额外的氨基酸取代。然而，“缺陷型”菌株被分为以下四种类型： 10 株在编码区含有终止密码子； 10 株在编码区内含有终止密码子；六株含有IS；一种在 C 末端结构域附近有短缺失的菌株；以及两个在测序区域没有终止密码子的菌株。使用OprD抗体的Western blot分析表明，OprD蛋白对“全长型”菌株的结合能力正常，而对“缺陷型”菌株的结合能力无一例外地丧失。这些结果表明，OprD 结构和碳青霉烯类药物的抗菌活性在铜绿假单胞菌中被证明是高度相关的。
A total of 99 clinical isolates of metallo-ss-lactamase-negative Pseudomonas aeruginosa collected in Japan between 1998 and 2001 were studied for their susceptibilities to carbapenem agents and corresponding oprD gene mutations. The OprD sequence of each strain was grouped into two major classes, based on the pattern of alterations. Eighty strains (80.8%) were so-called 'full length type', whose OprD proteins were fully encoded. The remaining 19 strains (19.2%) were so-called 'defective type', which possessed deletions or major alterations that might cause conformational changes in the OprD porin protein. The changes in 'defective type' strains led to 15-, 17- and 23-fold increases in the geometric mean MIC for imipenem, meropenem and biapenem compared with 'full length type' strains, respectively. 'Full length type' strains were further classified into six carbapenem susceptible types with the exception of four carbapenem-resistant subtypes with additional amino acid substitutions at D43, G183, R154, G314, G316. However, 'defective type' strains were classified into four types as follows: 10 strains which contained a stop codon within the coding region; six strains which contained IS; one strain with a short deletion near the C-terminal domain; and two strains without a stop codon in the sequenced region. Western blot analysis using OprD antibody showed that binding abilities of OprD proteins against 'full length type' strains were normal, whereas those against 'defective type' strains were lost without exception. These results indicate that OprD structure and antimicrobial activities for carbapenem agents proved to be highly correlated in P. aeruginosa.