Rapid and Specific Detection of the Escherichia coli Sequence Type 648 Complex within Phylogroup F
Rapid and Specific Detection of the Escherichia coli Sequence Type 648 Complex within Phylogroup F
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DOI：
10.1128/jcm.01949-16
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发表时间：
2017-04-01
影响因子：
9.4
通讯作者：
Gordon, David M.
中科院分区：
文献类型：
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作者：
Johnson, James R.;Johnston, Brian D.;Gordon, David M.
大肠杆菌序列648型复合体（STc648）是系统群f中的一个新兴谱系，以前包括在系统群d中，与多药耐药有关。在这里，我们设计并验证了一种新的基于多重pcr的STc648检测方法，该方法利用了(i)定义ST648的两个多位点序列分型等位基因icd等位基因96和gyrB等位基因87的四个独特的单核苷酸多态性；（ii） STc648中典型的缺失uidA，一种大肠杆菌特有的编码β -葡萄糖醛酸酶的基因。在包括109种STc648以外的来自a、B1、B2、C、D、E和F系群的212株菌株验证集中，该检测显示出100%的灵敏度（95%置信区间[CI]， 82%至100%）和特异性（95% CI， 98%至100%）。在两个相距遥远的实验室中，使用煮沸的裂解物或dnazol纯化的DNA作为模板DNA，它的功能相似。因此，这种新型的基于多重PCR的检测方法将使任何配备诊断PCR的实验室能够快速、准确、经济地筛选大肠杆菌分离株是否属于STc648。
The Escherichia coli sequence type 648 complex (STc648) is an emerging lineage within phylogroup F-formerly included within phylogroup D-that is associated with multidrug resistance. Here, we designed and validated a novel multiplex PCR-based assay for STc648 that took advantage of (i) four distinctive single-nucleotide polymorphisms in icd allele 96 and gyrB allele 87, two of the multilocus sequence typing alleles that define ST648; and (ii) the typical absence within STc648 of uidA, an E. coli-specific gene encoding beta-glucuronidase. Within a diverse 212-strain validation set that included 109 STs other than STc648, from phylogroups A, B1, B2, C, D, E, and F, the assay exhibited 100% sensitivity (95% confidence interval [CI], 82% to 100%) and specificity (95% CI, 98% to 100%). It functioned similarly well in two distant laboratories that used boiled lysates or DNAzol-purified DNA as the template DNA. Thus, this novel multiplex PCR-based assay should enable any laboratory equipped for diagnostic PCR to rapidly, accurately, and economically screen E. coli isolates for membership in STc648.