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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 four kinds of non-fermentative pathogens, Acinetobacter baumannii (Ab), Pseudomonas aeruginosa (Pa) Five quinolone resistance-related genes mediated by Stenotrophomonas maltophilia (Sm) and Chryseobacterium meningosepticum (Cm) plasmids provide basis for clinical rational use of quinolone antibacterials. Methods Nineteen clinical isolates (6 Ab, 2 multidrug-resistant, 4 pan-drug-resistant), 6 Strain Pa (3 strains with multi-drug resistance and pan-drug resistance), 4 strains with Sm (2 strains with multi-drug resistance and 2 strains with pan-resistance) and 3 strains with meningitis (2 strains with multiple drug resistance and 1 strain with pan-drug resistance) ) Were identified and susceptibility testing was performed. Five quinolone related genes mediated by quinolone resistant plasmids were detected by PCR, namely quinolone resistance A (qnrA), aminoglycoside acetyltransferase Aminoglycoside acetyltransferase ciprofloxacin resistance variant (acc (6 ’) - Ib-cr) and three integron genes (intI1,2,3) The amplified product was analyzed by 1% agarose gel electrophoresis and sequenced, and the bioinformatics methods were used for comparative analysis. Results Eight strains of acc (6 ’) - Ib-cr were carried in 19 strains of this strain, of which 6 strains were pan-resistant (Ab 3, Pa 2 and Sm 1) 10); 2 multi-resistant bacteria (Ab, Pa each one), multi-bacteria carrying rate of 22.0% (2/9). There was no significant difference in the carrying rates of pan-bacteria-resistant and multi-resistant bacteria (P <0.05). 11 strains of type Ⅰ integron were detected, of which 6 strains were pan-resistant (Ab3 strain, Pa2 strain and Sm1 strain) with a carriage rate of 60.0% (6/10). Five multi-drug resistant strains 1 strain). The carrying rate was 55.6% (5/9). There was no significant difference in the carrying rate of multidrug-resistant bacteria and the carrying rate of multi-resistant bacteria (P> 0.05). 6 strains were detected acc (6 ’) - Ib-cr and intI1, qnrA and intI2, intI3 were negative. Conclusion The quinolone resistance in clinical isolates of this group is related to acc (6 ’) - Ib-cr and intI1, but not to qnrA, intI 2and intI 3. The acc (6 ’) - Ib-cr carrier rate was higher than that of multi-bacteria, while the intI1 carrier rate had no significant difference. The detection rates of Ab and Pa were higher and similar in both genes.