本文選題：肺炎克雷伯菌 + 生物膜　； 參考：《中國(guó)抗生素雜志》2017年08期

【摘要】：目的分析耐碳青霉烯類(lèi)肺炎克雷伯菌(CRKPs)的生物膜形成能力與OqxAB外排泵的相關(guān)性。方法收集2008—2013年間溫州醫(yī)科大學(xué)附屬第一醫(yī)院臨床分離的碳青霉烯類(lèi)耐藥肺炎克雷伯菌37株。采用微量肉湯稀釋法檢測(cè)其對(duì)亞胺培南、厄他培南以及美羅培南的最低抑菌濃度(minimal inhibitory concentration,MIC),結(jié)晶紫染色法測(cè)定其生物膜形成能力,PCR方法篩查外排泵基因oqxA和oqxB的攜帶情況并分析亞抑菌濃度的外排泵抑制劑羰基氰氯苯腙(CCCP)對(duì)生物膜形成的影響;實(shí)時(shí)熒光定量PCR(qRT-PCR)檢測(cè)CCCP存在與不存在條件下生物膜狀態(tài)與浮游狀態(tài)oqxB外排泵基因的表達(dá)量。用統(tǒng)計(jì)學(xué)軟件SPSS 17.0分析實(shí)驗(yàn)數(shù)據(jù)。結(jié)果 37株肺炎克雷伯菌均對(duì)至少1種碳青霉烯類(lèi)藥物的敏感性降低,37株菌都具有不同程度的生物膜形成能力(A590范圍為0.044~1.113);在所有菌株中,只有7株菌存在oqxA或oqxB外排泵基因的缺失,與外排泵基因完整的試驗(yàn)菌株相比,這7株菌形成生物膜的能力較低;外排泵抑制試驗(yàn)發(fā)現(xiàn)亞抑菌濃度的CCCP對(duì)實(shí)驗(yàn)菌株的生物膜形成能力都有不同程度的促進(jìn)作用;qRT-PCR結(jié)果證明,生物膜狀態(tài)下菌株的外排泵基因的表達(dá)量要高于浮游狀態(tài)下的表達(dá)量。結(jié)論 OqxAB外排泵過(guò)表達(dá)可能會(huì)促進(jìn)肺炎克雷伯菌生物膜的形成。
[Abstract]:Objective to analyze the relationship between biofilm forming ability of Klebsiella pneumoniae (CRKPs) and OqxAB efflux pump. Methods 37 strains of carbapenem resistant Klebsiella pneumoniae isolated from the first affiliated Hospital of Wenzhou Medical University from 2008 to 2013 were collected. Determination of imipenem by broth dilution method, The minimum inhibitory concentrations of ertapenem and meropenem (minimal inhibitory concentration), the biofilm formation ability of ertapenem and meropenem were determined by crystal violet staining. PCR was used to screen for the carrying of efflux pump gene oqxA and oqxB, and to analyze the carbonyl of efflux pump inhibitor of subinhibitory concentration. The effect of CCCP on the formation of biofilm; Real-time fluorescence quantitative PCR (qRT-PCR) was used to detect the expression of oqxB efflux pump gene in the presence and absence of CCCP. Statistical software SPSS 17.0 was used to analyze the experimental data. Results the sensitivity of 37 strains of Klebsiella pneumoniae to at least one carbapenes was decreased. 37 strains of Klebsiella pneumoniae showed different biofilm forming ability (A590 range: 0.044% 1.113). Only 7 strains had the absence of oqxA or oqxB efflux pump genes. Compared with the experimental strains with intact efflux pump genes, the biofilm formation ability of these 7 strains was lower. The results of efflux pump inhibition test showed that CCCP with subinhibitory concentration could promote the biofilm formation ability in different degree. The results of qRT-PCR showed that the expression of efflux pump gene in biofilm was higher than that in planktonic state. Conclusion the overexpression of OqxAB efflux pump may promote the biofilm formation of Klebsiella pneumoniae.
【作者單位】： 溫州醫(yī)科大學(xué)附屬第一醫(yī)院醫(yī)學(xué)檢驗(yàn)中心;浙江醫(yī)院檢驗(yàn)科;溫州醫(yī)科大學(xué)檢驗(yàn)醫(yī)學(xué)院生命科學(xué)學(xué)院;
【基金】：浙江省衛(wèi)生高層次創(chuàng)新人才培養(yǎng)工程項(xiàng)目(No.浙衛(wèi)發(fā)[2012]241號(hào))
【分類(lèi)號(hào)】：R446.5

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