[1]朱春晖),王文杏),陈 强),等.鼠伤寒沙门菌质粒毒力基因对耐药表型的影响[J].郑州大学学报(医学版),2018,(02):192-198.[doi:10.13705/j.issn.1671-6825.2017.08.080]
 ZHU Chunhui),WANG Wenxing),CHEN Qiang),et al.Influence of spv on antibiotic resistance of Salmonella typhimurium[J].JOURNAL OF ZHENGZHOU UNIVERSITY(MEDICAL SCIENCES),2018,(02):192-198.[doi:10.13705/j.issn.1671-6825.2017.08.080]
点击复制

鼠伤寒沙门菌质粒毒力基因对耐药表型的影响()
分享到:

《郑州大学学报(医学版)》[ISSN:1671-6825/CN:41-1340/R]

卷:
期数:
2018年02期
页码:
192-198
栏目:
应用研究
出版日期:
2018-03-20

文章信息/Info

Title:
Influence of spv on antibiotic resistance of Salmonella typhimurium
作者:
朱春晖1)王文杏2)陈 强2)陈庆法2)吴春雪2)刘晓艳2)赖华毅2)余晓君3)
1)江西省儿童医院感染科 南昌 330006
2)江西省儿童医院内科 南昌 330006
3)江西省儿童医院检验科 南昌 330006
Author(s):
ZHU Chunhui1)WANG Wenxing2)CHEN Qiang2)CHEN Qingfa2)WU Chunxue2)LIU Xiaoyan2)LAI Huayi2)YU Xiaojun3)
1)Department of Infectious Disease,the Children's Hospital of Jiangxi Province, Nanchang 330006
2)Department of Internal Medicine,the Children's Hospital of Jiangxi Province, Nanchang 330006
3)Department of Clinical Laboratory,the Children's Hospi
关键词:
鼠伤寒沙门菌 沙门菌质粒毒力基因 抗生素耐药 腹泻
Keywords:
Salmonella typhimurium spv antibiotic resistance diarrhea
分类号:
R378.2
DOI:
10.13705/j.issn.1671-6825.2017.08.080
摘要:
目的:探讨沙门菌质粒毒力基因(spv)表达改变对鼠伤寒沙门菌耐药表型的影响。方法:在eGFP基因序列上下游增加启动子及终止子并在两端位点上、下游设计酶切位点,以pACYC184-eGFP为载体,构建spv基因回补质粒,电转化至前期构建的鼠伤寒沙门菌spvB及spvC基因共同缺陷株获得基因回补株,检测菌株间耐药性的差异及acrAB表达的差异。结果:成功构建鼠伤寒沙门菌spvB、spvC基因荧光回补株,连续培养12代质粒未发生丢失。与野生型菌株及回补株相比,spv基因缺陷变异株对哌拉西林他唑巴坦、头孢噻肟、头孢西丁、四环素、氯霉素及环丙沙星的MIC值下降。spv基因缺陷变异株AcrA周质蛋白水平低于野生菌株及回补株(P<0.05)。结论:在spvB及spvC基因共同缺陷鼠伤寒沙门菌基础上的基因回补株构建成功,spv下调能改变鼠伤寒沙门菌的耐药表型,这可能是通过影响外排泵AcrA来完成的。
Abstract:
Aim: To observe the influence of spv on the antibiotic resistance of Salmonella typhimurium isolated from the faeces of the diarrhea children.Methods: The promoter was added to the upstream and the terminator was added to the downstream of the sequence of eGFP published in GeneBank as well as restriction sites was designed,and finally pACYC184-eGFP was synthesized using multiple PCR.The spv-complementing plasmid was constructed using pACYC184-eGFP vector,then electroporated into the spvB and spvC gene-deleted mutant to acquire the corresponding strain, and then drug resistance and the expression of resistance related genes of different strains were examined.Results: PCR and gene sequencing confirmed successful construction of Salmonella typhimurium spvB, spvC gene complemented mutant strain with fluorescence. Low copy plasmid does not lose after 12 generation continuous culture. Compared with the wide type strain and the compensating strain,the MICs of spvB and spvC gene-deleted mutant strain descended original MIC values for piperacillin/tazobactam, cefotaxime, cefoxitin, tetracycline, chloramphenicol and ciprofloxacin.The expression of periplasmic protein AcrA in spvB and spvC gene-deleted mutant strain was lower than that of wide type strain and the compensating strain(P<0.05).Conclusion: The spv complementation strain has been constructed successfully on the basis of spvB and spvC-deleted mutant strain,the downregulation of spv can change the antibiotic resistance phenotype of Salmonella typhimurium,which may due to the downregulation of AcrA.

参考文献/References:

[1] BRENT AJ,OUNDO JO,MWANGI I,et al.Salmonella bacteremia in Kenyan children[J].Pediatr Infect Dis,2006,25(3):230
[2] 陈强,余晓君,李俏俏,等.引起儿童腹泻的沙门菌属临床分离株的耐药特点及分子流行病学研究[J].中华检验医学杂志,2011,34(3):249
[3] 吴春雪,陈强,李红,等.鼠伤寒沙门菌spvB基因缺陷变异株的制备及其抗酸能力检测[J].华中科技大学学报(医学版),2014,43(4):399
[4] 陈强,吴春雪,余晓君,等.鼠伤寒沙门菌spvC基因缺陷变异株的制备[J].江苏大学学报(医学版),2014,24(3):235
[5] 刘晓艳,陈强,李红,等.spvB/spvC基因对沙门菌毒力及宿主免疫功能的影响[J].南方医科大学学报,2015,35(11):1649
[6] FISHER IST.International trends in Salmonella serotypes 1998-2003: a surveillance report from the enter-net internationalsurveillance network[J].Euro Surveill 2004,9(11):9
[7] HARVEY RR,FRIEDMAN CR,CRIM SM,et al.Epidemiology of Salmonella enterica serotype dublin infections among Humans, United States, 1968-2013[J].Emerg Infect Dis,2017,23(9):1493
[8] FIELD CM,SUMMERS DK.Multicopy plasmid stability:revisiting the dimer catastrophe[J].J Theor Biol,2011,291:119
[9] LOBATO-MÁRQUEZ D,MOLINA-GARCÍA L,MORENO-CÓRDOBA I,et al.Stabilization of the virulence plasmid pSLT of Salmonella typhimurium by three maintenance systems and its evaluation by using a new stability test[J].Front Mol Biosci,2016,17(3):66
[10]GONG CC,KLUMPP S.MODELING sRNA-regulated plasmid maintenance[J].PLoS One,2017,12(1):e0169703
[11]ZAHARIK ML,VALLANCE BA,PUENTE JL,et al.Host-pathogen interactions:host resistance factor Nramp1 up-regulates the expression of Salmonella pathogenicity island-2 virulence genes[J].Proc Natl Acad Sci USA,2002,99(24):15705
[12]BUCKNER MM,BLAIR JM,LA RAGIONE RM,et al.Beyond antimicrobial resistance:evidence for a distinct role of the AcrD efflux pump in Salmonella biology[J].MBio,2016,7(6):16
[13]CHEN Y,HU DX,ZHANG QJ,et al.Efflux pump overexpression contributes to tigecycline heteroresistance in Salmonella enterica serovar Typhimurium[J].Front Cell Infect Microbiol,2017,7:37
[14]WU SY,WANG LD,LI JL,et al.Salmonella spv locus suppresses host innate immune responses to bacterial infection[J].Fish Shellfish Immunol,2016,58:387
[15]黄瑞,吴淑燕,张学光.伤寒杆菌耐药质粒pRsags毒力基因的研究[J].中华微生物学和免疫学杂志,2003,23(5):393
[16]WESTON N,SHARMA P,RICCI V,et al.Regulation of the AcrAB-TolC efflux pump in Enterobacteriaceae[J].Res Microbiol,2017.doi:10.1016/j.resmic.2017.10.005

备注/Memo

备注/Memo:
【基金项目】江西省自然科学基金资助项目(20132BAB205036); 江西省科技支撑计划课题资助项目(20111BBG70015-2)
【作者简介】陈强,通信作者,女,1961年1月生,教授,研究方向:儿童感染性疾病的诊治,E-mail:jx_cq@163.com
更新日期/Last Update: 2018-03-20