背景:本研究的目的是研究质粒介导的喹诺酮耐药(PMQR)基因的存在和生物膜的形成在几种对喹诺酮耐药的临床志贺氏菌分离株。
方法:本横断面研究(2020年11月至2021年12月)收集了150名腹泻患者(10岁以下)的粪便样本。在Hektoen肠琼脂和木糖赖氨酸脱氧胆酸盐琼脂上培养样品后,标准微生物学测试,VITEK2系统,和聚合酶链反应(PCR)用于鉴定志贺氏菌分离株。肉汤微量稀释法用于确定抗生素敏感性。PMQR基因包括qnrA,qnrB,qnrC,qnrD,qnrE,qnrS,qnrVC,qepA,OQXAB,aac(6\')-Ib-cr,并通过PCR和微量滴定板法研究了耐喹诺酮类药物分离株的CRPP和生物膜形成,分别。使用肠细菌重复基因间共有聚合酶链反应(ERIC-PCR)技术确定喹诺酮耐药分离株的克隆相关性。
结果:共有95株志贺氏菌分离株,包括S.sonnei(53,55.8%),S、flexneri(39,41.1%),和鲍迪氏链球菌(3,3.2%)被鉴定。分离株对氨苄青霉素的耐药率最高(92.6%,n=88/95)。总的来说,95个分离株中的42个(44.2%)同时对两种或更多种喹诺酮类药物具有抗性,包括26个(61.9%)S.sonnei和16个(38.1%)flexneri。所有分离株均具有多重耐药(对3种以上抗生素耐药)。PMQR基因的发生如下:qnrS(52.4%),qnrA和ac(6')-Ib-cr(33.3%),和qnrB(19.0%)。物种患病率如下:61.5%和37.5%(qnrS),19.2%和56.3%(qnrA),38.5%和25.0(ac(6')-Ib-cr),桑内和福内分别为19.2%和18.8%(qnrB),分别。未检测到其他PMQR基因。总的来说,52.8%(28/53)的喹诺酮敏感株和64.3%(27/42)的喹诺酮耐药株是生物膜生产者。喹诺酮耐药和喹诺酮敏感分离株之间的生物膜形成没有显着差异(P值=0.299)。根据ERIC-PCR,耐喹诺酮类药物的分离株表现出很高的遗传多样性。
结论:似乎qnrS,qnrA,aac(6')-Ib-cr在本地区志贺氏菌分离株的喹诺酮耐药中起重要作用。此外,耐喹诺酮的福氏杆菌和松内分离株具有很高的遗传多样性。因此,抗生素治疗需要根据监测结果进行常规修订.
BACKGROUND: The purpose of this study was to look into the presence of plasmid-mediated quinolone resistance (PMQR) genes and biofilm formation in several species of clinical Shigella isolates that were resistant to quinolones.
METHODS: The stool samples of 150 patients (younger than 10 years) with diarrhea were collected in this cross-sectional study (November 2020 to December 2021). After cultivation of samples on Hektoen Enteric agar and xylose lysine deoxycholate agar, standard microbiology tests, VITEK 2 system, and polymerase chain reaction (PCR) were utilized to identify Shigella isolates. The broth microdilution method was used to determine antibiotic susceptibility. PMQR genes including qnrA, qnrB, qnrC, qnrD, qnrE, qnrS, qnrVC, qepA, oqxAB, aac(6\')-Ib-cr, and crpP and biofilm formation were investigated in quinolone-resistant isolates by PCR and microtiter plate method, respectively. An enterobacterial repetitive intergenic consensus polymerase chain reaction (ERIC-PCR) technique was used to determine the clonal relatedness of quinolone-resistant isolates.
RESULTS: A total of 95 Shigella isolates including S. sonnei (53, 55.8%), S. flexneri (39, 41.1%), and S. boydii (3, 3.2%) were identified. The highest resistance rates of the isolates were against ampicillin (92.6%, n = 88/95). Overall, 42 of 95 (44.2%) isolates were simultaneously resistant against two or more quinolones including 26 (61.9%) S. sonnei and 16 (38.1%) S. flexneri. All isolates were multidrug-resistant (resistance to more than 3 antibiotics). The occurrence of PMQR genes was as follows: qnrS (52.4%), qnrA and aac(6\')-Ib-cr (33.3%), and qnrB (19.0%). The prevalence in species was as follows: 61.5% and 37.5% (qnrS), 19.2% and 56.3% (qnrA), 38.5% and 25.0 (aac(6\')-Ib-cr), and 19.2% and 18.8% (qnrB) for S. sonnei and S. flexneri, respectively. The other PMQR genes were not detected. In total, 52.8% (28/53) of quinolone-susceptible and 64.3% (27/42) of quinolone-resistant isolates were biofilm producers. Biofilm formation was not significantly different between quinolone-resistant and quinolone-susceptible isolates (P-value = 0.299). Quinolone-resistant isolates showed a high genetic diversity according to the ERIC-PCR.
CONCLUSIONS: It seems that qnrS, qnrA, and aac(6\')-Ib-cr play a significant role in the quinolone resistance among Shigella isolates in our region. Also the quinolone-resistant S. flexneri and S. sonnei isolates had a high genetic diversity. Hence, antibiotic therapy needs to be routinely revised based on the surveillance findings.