摘要
目的检测2种耐药表型(多耐和泛耐)的4种非发酵病原菌,,鲍曼不动杆菌(Acinetobacter baumannii,Ab)、铜绿假单胞菌(Pseudomonas aeruginosa,Pa)、嗜麦芽窄食单胞菌(Stenotrophomonas maltophilia,Sm)和脑膜炎金黄杆菌(Chryseobacterium meningosepticum,Cm)质粒介导的5种喹诺酮类耐药相关基因,为临床合理使用喹诺酮类抗菌药物提供依据。方法应用Phoenix NMIC/ID-109鉴定/药敏板和API 20NE鉴定条/PSE5.0药敏条对19株临床分离菌(6株Ab(多耐药2株、泛耐药4株)、6株Pa(多耐药、泛耐药各3株)、4株Sm(多耐药、泛耐药各2株)和3株脑膜炎金黄杆菌(2株多耐药、1株泛耐药))进行鉴定和药敏试验,并用PCR法检测其喹诺酮类耐药质粒介导的5种喹诺酮类相关基因,即喹诺酮类药物耐药相关基因A(quinolone resistance A,qnrA)、氨基糖苷类乙酰转移酶基因acc(6')-Ib的环丙沙星耐药变异体基因(aminoglycoside acetyltransferase ciprofloxacin resistance variant,acc(6')-Ib-cr)及3种整合子基因(intI1、2、3),扩增产物经1%琼脂糖电泳分析并测序,利用生物信息学的方法进行比对分析。结果本组19株菌中携带acc(6')-Ib-cr 8株,6株为泛耐药(Ab 3株,Pa 2株,Sm 1株),泛耐菌携带率60.0%(6/10);2株多耐菌(Ab、Pa各1株),多耐菌携带率22.0%(2/9)。泛耐菌携带率与多耐菌携带率比较差异无统计学意义(P<0.05)。11株检出Ⅰ类整合子,其中6株泛耐药(Ab3株,Pa2株,Sm1株),携带率60.0%(6/10);5株多耐药菌(Pa3株,Ab、Cm各1株),携带率55.6%(5/9),泛耐菌携带率与多耐菌携带率比较差异无统计学意义(P>0.05)。6株同时检出acc(6')-Ib-cr和intI1,qnrA和intI2、intI3均阴性。结论本组临床分离菌喹诺酮类药物耐药与acc(6')-Ib-cr和intI1有关,与qnrA,intI 2and intI 3无关。acc(6')-Ib-cr携带率泛耐菌高于多耐菌,而intI1携带率无显著差异;2种基因在Ab和Pa的检出率较高且相近。
Objective To detect 5genes associated with plasmid-mediated quinolone resistance（PMQR）in two drug-resistant phenotypes（multi-resistant（MR）and pan-resistant（PR））of four non-fermenting Gram-negative bacteria,Acinetobacter baumannii,Pseudomonas aeruginosa,Stenotrophomonas maltophilia,and Chryseobacterium meningosepticum（Cm）,in order to provide a rational basis for clinical use of quinolones. Methods The susceptibility of 4non-fermenting Gram-negative bacteria to antimicrobial agents（expressed as the MIC）was determined using the Phoenix NMIC/ID-109 panel and bioMerieux API 20 NE ID panel/PSE5.0panel.PCR was used to detect 5genes associated with PMQR（quinolone resistance A（qnrA）,aminoglycoside acetyltransferase ciprofloxacin resistance variant（acc（6＇）-Ib-cr）,and 3integrons（intI1,2,3））in 19 strains of non-fermenting Gram-negative bacteria（6strains of A.baumannii（4PR strains and 2 MR strains）,6strains of P.aeruginosa（3PR strains and 3 MR strains）,4strains of S.maltophilia（2PR strains,2MR strains）,and 3strains C.meningosepticum（1PR strains,2MR strains））.Results were verified with gel electrophoresis and DNA sequencing using bioinformatic analysis tools. Results Of the 19 strains,8strains（6PR strains and 2 MR strains）were positive for acc（6＇）-Ib-cr.PR strains were positive for acc（6＇）-Ib-cr at a higher rate（60%,6/10strains）than MR strains were（22.0%,2/9strains：1 A.baumannii strain,1 P.aeruginosastrain）（P0.05）.Eleven strains（6PR strains：3 A.baumannii strains,2 P.aeruginosastrains,and 1 S.maltophiliastrain,5MR strains：1 A.baumannii strain,3 P.aeruginosastrains,and 1 C.meningosepticumstrain）were positive for intI1.PR strains were positive for intI1（60.0%,6/10strains）at about the same rate as MR strains（55.6%,5/9strains）（P0.05）.Six strains（4PR strains and 2 MR strains）were positive for acc（6＇）-Ib-cr and intI1.Two PR strains（1 P.aeruginosastrain and 1 S.maltophiliastrain）were positive for acc（6＇）-Ib-cr alone,and 5strains（1 P.aeruginosa PR strain,2 P.aeruginosa MR strains,1 S.maltophilia PR strain,and 1 C.meningosepticum MR strain）were positive for intI1 alone.No strains were positive for qnrA,intI2,or intI3. Conclusion The mechanisms of quinolone resistance of the 19 strains were related to acc（6＇）-Ib-cr and intI1 but not to qnrA,intI2,or intI3.PR isolates tested positive for acc（6＇）-Ib-cr at a higher rate than MR isolates.There were no differences between PR isolates and MR isolates in terms of their positivity for intI1.A.baumannii and P.aeruginosatested positive for acc（6＇）-Ib-cr and intI1 at a high rate.
出处
《中国病原生物学杂志》
CSCD
北大核心
2016年第5期464-467,共4页
Journal of Pathogen Biology
基金
福建省自然科学基金项目(No.2015J01514)
泉州市社会发展领域科技计划项目(No.2014Z24)
泉州医学高等专科学校'国家骨干院校建设'重点资助科研项目(No.2014XJ1313)
