Genotypic Diversity and Characterization of Quinolone Resistant Determinants from Enterobacteriaceae in Yaoundé, Cameroon

Background: Enterobacteriaceae causes many types of infections which are often treated with quinolones and fluoroquinolone (Q/FQ). The resistance mechanisms to Q/FQ are usually associated with mutations in the quinolone resistance determining region which alter the conformation of target amino acid residues within the protein and in the qnr genes. This study aimed at determining the antimicrobial resistant profile of a sample of Enterobacteriaceae from Cameroon and the genetic diversity in quinolone-resistant isolates in view of implementing a better management, treatment, control and prevention of the transmission of these resistant strains. Methods: Identification and antimicrobial susceptibility testing was done using VITEK 2. The detection of plamid-mediated quinolone resistance (PMQR) genes was carried out using the conventional PCR method. Sequencing was done using the Applied Biosystem 3500 genetic analyser. DNA fingerprint was obtained using Pulsed-Field Gel electrophoresis. Results: Among 440 Enterobacteriaceae, the most prevalent genera were: Escherichia 178/440 (39.5%);Klebsiella 148/440 (33.6%);Enterobacter 35/440 (8%);Pantoea 28/440 (6.4%);Proteus 14/440 (3.2%) Salmonella 13/440 (3%). Ampicillin resistance showed the highest prevalence with 371/440 (81%) and Imipenem the lowest resistance 9/440 (2.1%). The ciprofloxacin resistance rate was 161/440 (36.6%). The detected plasmid mediated quinolone resistance (PMQR) genes were: qnrA, 2/161 (1.2%);qnrB, 31/161 (19.3%);qnrS, 13/161 (8.1%): Aac (6')Ib-cr, 84/161 (52.2%) and qepA, 3/161 (1.9%). There were several mutations in the parC gene of Klebsiella leading to S80D and S80N substitutions. Two pairs of Klebsiella peumoniae strains were phenotypically and genotypically identical with 100% similarity in the dendrogramme. Conclusion: This study showed that quinolone resistance was high. The PMQR genes contributing to this resistance were diverse. This high PMQR indicates that there has been an unknown circulation of these genes in our community. To avoid the rapid dissemination of these PMQR genes continuous surveillance of antimicrobial resistance should be carried out not only in humans but also in animals to monitor the evolution of these genes.

兔源大肠杆菌对喹诺酮药物耐药性及质粒介导的耐药基因检测

为了解四川省兔源大肠杆菌(E.coli)对喹诺酮药物耐药性及质粒介导的喹诺酮耐药(PMQR)基因的携带情况,本研究从四川地区规模化家兔养殖场共分离鉴定出97株E.coli,采用Kirby-Bauer(K-B)纸片法对分离株进行喹诺酮药物耐药性测定,同时采用PCR方法对qnrA、qnrB、qnrC、qnrD、qnrS、qnrVC、aac(6')-Ib-cr、qep A、oqx A、oqx B等PMQR基因进行检测。结果显示:分离株对左氧氟沙星耐药率最高为49.48%,对依诺沙星、诺氟沙星、恩诺沙星、氧氟沙星和环丙沙星的耐药率依次为48.45%、46.39%、39.17%、35.05%和34.02%;PMQR检测发现:aac(6')-Ib-cr检出率最高为80.4%,qnrD、qnrS、oqxA和oqxB,检出率分别为59.8%、59.8%、63.9%和51.5%,未检出qnrA、qnrB、qnrC、qnrVC和qepA。本研究通过对97株兔源E.coli对喹诺酮药物的耐药性及相关PMQR基因检测,初步明确四川地区兔源大肠杆菌对喹诺酮药物的耐药情况及PMQR的流行情况,为该地区家兔大肠杆菌病的防治中有效使用喹诺酮类药物提供参考。

猪源携带质粒介导喹诺酮类耐药基因大肠杆菌气源性传播的研究

为了调查养猪场携带质粒介导喹诺酮类耐药基因大肠杆菌的气源性传播情况,本试验分别在5个猪场舍内及舍外上风向和下风向不同距离收集空气样品,并在舍内随机采集粪便样品,分离大肠杆菌。药敏试验检测其对14种抗生素的耐药性。以质粒介导喹诺酮类耐药基因(qnr、aac(6′)-Ib-cr、qepA)为指示基因,利用肠杆菌基因间重复一致序列聚合酶链式反应(ERIC-PCR)鉴定技术,分别对5个猪场不同样品中大肠杆菌的遗传相似性进行分析,评估其向舍外空气的传播情况。结果显示,5个猪场中的大肠杆菌对庆大霉素、卡那霉素、四环素、链霉素、萘啶酸、复方新诺明等12种常用抗生素耐药率较高,且呈现多重耐药。ERIC-PCR结果显示,46.34%(19/41)的舍内空气分离株与粪便分离株来源相同,其中73.68%(14/19)的分离株携带的耐药基因也相同;68.42%(26/38)的舍外空气分离株与舍内空气或粪便分离株来源相同,其中65.38%(17/26)的菌株携带相同的耐药基因。结果表明,起源于舍内粪便的携带质粒介导喹诺酮类耐药基因的大肠杆菌能形成气溶胶污染舍内空气,并借助舍内外气体交换,传播到舍外不同距离空气中(≥400m),对养殖场周围的环境卫生及社区居民的健康形成威胁。

犬源大肠杆菌质粒介导的喹诺酮类药物耐药基因的检测及药物敏感性分析

采用PCR方法检测了102株犬源大肠杆菌中质粒介导的喹诺酮类药物耐药(plasmid-mediated quinolone resistance,PMQR)基因的分布,并采用微量肉汤稀释法测定了6种抗菌药物对犬源大肠杆菌的最小抑菌浓度(MIC)。在被检测的8种PMQR基因中,仅有oqx A、oqx B、qnr S这3种耐药基因被检测出,检出率分别为9.80%、11.76%和14.70%,其中同时含有2种耐药基因的菌株检出率为8.82%,同时含有3种耐药基因的菌株检出率为4.90%;犬源大肠杆菌对喹诺酮类药物已经具有较高的耐药性,其对恩诺沙星、环丙沙星和氧氟沙星的耐药率分别达到了71.57%、64.71%、48.04%。