Ragimbeau Catherine, Colin Stephanie, Devaux Anthony, Decruyenaere Frédéric, Cauchie Henry-Michel, Losch Serge, Penny Christian, Mossong Joël
National Health Laboratory, Surveillance and Epidemiology of Infectious Diseases, 1 rue Louis Rech, Dudelange L-3555, Luxembourg.
BMC Microbiol. 2014 Aug 28;14:205. doi: 10.1186/s12866-014-0205-7.
Surveillance and field investigations of Campylobacter infections require molecular tools with genetic markers appropriate for tracing purposes, i.e. based on the principle that some Campylobacter lineages acquire a host signature under adaptive selection pressure. We developed a sequence-based method targeting the quinolone resistance determining region within the subunit A of DNA gyrase (gyrA). Host specificity was evaluated by characterizing two collections of Campylobacter jejuni (N = 430) and Campylobacter coli (N = 302) originating from surface waters, domestic mammals and poultry.
Based on nucleotide identity, a total of 80 gyrA alleles were observed. Thirty nine alleles assigned to C. coli encoding two peptides fell into three clades: two associated with surface waters and one associated with domestic mammals and poultry. The variability in GC content generated by synonymous mutations suggested that surface waters isolates originated from two distinct ecological niches. A total of 42 alleles were recorded from C. jejuni strains and encoded 8 peptides including one lying in a distinct lineage associated with wildlife. Seven of the 23 alleles encoding peptide #1 displayed the synonymous mutation G408A not identified in poultry isolates. By contrast, the substitution Ser22Gly observed in 4 different peptide groups was significantly associated with domestic birds (P = 0.001). The change in amino acid sequences Thr86Ile conferring resistance to quinolones was significantly associated with poultry (P < 0.001) in both C. jejuni and C. coli with 38.7% and 67.9% of quinolone-resistant strains, respectively.
The gyrA typing method presented here is an informative tool as sequences appear to be predictive of particular ecological niches. Combined with multi-locus sequence typing, it could increase the resolution of source attribution, and combined with porA/flaA typing it could be suitable for detecting temporal clusters of human cases. All gyrA alleles identified were deposited in the freely accessible online database http://pubmlst.org/campylobacter.
弯曲杆菌感染的监测和现场调查需要具备适合溯源目的的遗传标记的分子工具,即基于某些弯曲杆菌谱系在适应性选择压力下获得宿主特征这一原理。我们开发了一种基于序列的方法,靶向DNA促旋酶亚基A(gyrA)内的喹诺酮耐药决定区。通过对来自地表水、家养哺乳动物和家禽的空肠弯曲杆菌(N = 430)和结肠弯曲杆菌(N = 302)的两个样本集进行特征分析,评估了宿主特异性。
基于核苷酸同一性,共观察到80个gyrA等位基因。分配给编码两种肽的结肠弯曲杆菌的39个等位基因分为三个进化枝:两个与地表水相关,一个与家养哺乳动物和家禽相关。同义突变产生的GC含量变异性表明,地表水分离株源自两个不同的生态位。从空肠弯曲杆菌菌株中记录到总共42个等位基因,编码8种肽,其中一种位于与野生动物相关的独特谱系中。编码肽#1的23个等位基因中有7个显示出家禽分离株中未发现的同义突变G408A。相比之下,在4个不同肽组中观察到的Ser22Gly替代与家禽显著相关(P = 0.001)。在空肠弯曲杆菌和结肠弯曲杆菌中,赋予喹诺酮抗性的氨基酸序列Thr86Ile的变化分别与38.7%和67.9%的喹诺酮耐药菌株的家禽显著相关(P < 0.001)。
本文介绍的gyrA分型方法是一种信息丰富的工具,因为序列似乎可以预测特定的生态位。与多位点序列分型相结合,它可以提高溯源的分辨率,与porA/flaA分型相结合,它可能适用于检测人类病例的时间聚集。所有鉴定出的gyrA等位基因都存放在可免费访问的在线数据库http://pubmlst.org/campylobacter中。