Objectives: This study evaluated the diagnostic performance of the eazyplex® SuperBug CRE (eSBCRE) system, based on a loop-mediated isothermal amplification (LAMP), for the detection of the most common extended-spectrum beta-lactamases (ESBL) and carbapenemase genes in 140 clinical isolates of Escherichia coli. Materials and Methods: ESBL (blaCTX-M-1group and blaCTX-M-9group) and carbapenemase (blaKPC, blaVIM, blaNDM, blaOXA-48, and blaOXA-181) genes were detected using the eSBCRE test and compared with the results obtained by PCR, real-time PCR, and phenotypic methods. Results: Concordant results of 100% between PCR/real-time PCR and eSBCRE assays were observed. Two of 140 E. coli isolates were positive for both ESBL and carbapenemase genes according to eSBCRE, PCR, and real-time PCR assays, whereas they were negative in double-disk synergy test. Of 16 E. coli isolates suspected of producing carbapenemase, 9 were positive for 48-oxacillinases (OXA-48) by 30 μg temocillin test, whereas the blaOXA-48 was found only in 1 E. coli isolate by all molecular methods. Maximum threshold time values (minutes:seconds) in the eSBCRE test were 6:00, 11:15, 11:00, and 9:00 for the blaCTX-M-1group, blaCTX-M-9group, blaVIM, and blaOXA-48 genes, respectively. Conclusions: The eSBCRE test based on LAMP method is a reliable, easy-to-use, and timesaving molecular system, which can be successfully used in the routine diagnostic for the rapid detection of the most common ESBL and carbapenemase genes among clinical E. coli isolates.

译文


目标:
这项研究基于回路介导的等温扩增(LAMP),评估了eazyplex®SuperBug CRE(eSBCRE)系统的诊断性能,以检测140种最常见的广谱β-内酰胺酶(ESBL)和碳青霉烯酶基因大肠杆菌的临床分离株。
材料和方法:
使用eSBCRE测试检测了ESBL(blaCTX-M-1组和blaCTX-M-9组)和碳青霉烯酶(blaKPC,blaVIM,blaNDM,blaOXA-48和blaOXA-181)基因,并将其与PCR结果进行了比较,时间PCR和表型方法。
结果:
观察到PCR /实时PCR和eSBCRE分析之间100%的一致结果。根据eSBCRE,PCR和实时PCR分析,在140株大肠杆菌中,有2株ESBL和碳青霉烯酶基因均为阳性,而在双盘协同试验中则为阴性。在30μg替莫西林试验中,在怀疑产生碳青霉烯酶的16株大肠杆菌中,有9株对48-氧杂环菌酶(OXA-48)呈阳性,而通过所有分子方法仅在1株大肠杆菌中发现了blaOXA-48。对于blaCTX-M-1组,blaCTX-M-9组,blaVIM和blaOXA-48,eSBCRE测试中的最大阈值时间值(分钟:秒)为6:00、11:15、11:00和9:00。基因分别。
结论:
基于LAMP方法的eSBCRE测试是可靠,易于使用且省时的分子系统,可成功用于常规诊断中,以快速检测临床大肠杆菌分离物中最常见的ESBL和碳青霉烯酶基因。

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