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首页> 外文期刊>感染症学雑誌 >Development and evaluation of novel loop-mediated isothermal amplification for rapid detection of bla(IMP-1) and bla(VIM-2) genes
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Development and evaluation of novel loop-mediated isothermal amplification for rapid detection of bla(IMP-1) and bla(VIM-2) genes

机译：快速检测bla（IMP-1）和bla（VIM-2）基因的新型环介导等温扩增的开发和评估

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摘要

Loop-mediated isothermal amplification (LAMP) amplifies a target gene with high specificity and rapidity under isothermal conditions. LAMP assays were developed for the rapid detection of metallo-beta-lactamase (MBL) genes such as bla(IMP-1)) and bla(VIM-2). We initially designed specific primers to detect MBL genes for LAMP assays and evaluated the specificity and sensitivity of these assays. LAMP assays amplified MBL genes under a constant temperature of 63 degrees C within 1 hour, and were compared to PCR in MBL-producing strains. The results of MBL genes typing by LAMP assays agree completely with PCR results. The lower detection limits of bla(IMP-1)- and bla(VIM-2)-LAMP assays using real-time turbidimeters were 30cfu/test and 3cfu/test. After amplification, products were directly observed by the naked eye with a fluorescent detection reagent. In conclusion, LAMP assays are convenient, rapid, and fully feasible for detecting MBL genes in ordinary clinical microbiology laboratories without special apparatus.
机译：环介导的等温扩增（LAMP）在等温条件下以高特异性和快速性扩增靶基因。开发了LAMP测定法以快速检测金属β-内酰胺酶（MBL）基因，例如bla（IMP-1）和bla（VIM-2）。我们最初设计了特异性引物来检测用于LAMP分析的MBL基因，并评估了这些分析的特异性和敏感性。 LAMP分析在1小时内于63摄氏度的恒定温度下扩增了MBL基因，并将其与产生MBL的菌株中的PCR进行了比较。通过LAMP分析进行MBL基因分型的结果与PCR结果完全一致。使用实时浊度计的bla（IMP-1）-和bla（VIM-2）-LAMP分析的最低检测限为30cfu / test和3cfu / test。扩增后，用荧光检测试剂直接用肉眼观察产物。总之，在普通的临床微生物学实验室中，无需特殊设备即可进行LAMP检测方便，快速且完全可行，可用于检测MBL基因。

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