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中华实验和临床感染病杂志(电子版) ›› 2017, Vol. 11 ›› Issue (06) : 556 -560. doi: 10.3877/cma.j.issn.1674-1358.2017.06.007

临床论著

志贺菌临床分离株AcrAB-TolC外排泵mRNA转录水平与氟喹诺酮耐药的相关性
刘奉云1, 张豪杰1, 杨贤1, 吴晓妹1, 祁伟1,()   
  1. 1. 300211 天津,天津医科大学第二医院感染性疾病研究所
  • 收稿日期:2014-02-27 出版日期:2017-12-15
  • 通信作者: 祁伟
  • 基金资助:
    天津市卫生局科技基金项目(No. 201440296)

Relationship between the mRNA transcription levels of AcrAB-TolC efflux pump in Shigella from clinical source and fluoroquinolone resistance

Feng Yun Liu1, Haojie Zhang1, Xian Yang1, Xiaomei Wu1, Wei Qi1,()   

  1. 1. Institute of Infectious Diseases, The Second Hospital of Tianjin Medical Univercity, Tianjin 300211, China
  • Received:2014-02-27 Published:2017-12-15
  • Corresponding author: Wei Qi
引用本文:

刘奉云, 张豪杰, 杨贤, 吴晓妹, 祁伟. 志贺菌临床分离株AcrAB-TolC外排泵mRNA转录水平与氟喹诺酮耐药的相关性[J]. 中华实验和临床感染病杂志(电子版), 2017, 11(06): 556-560.

Feng Yun Liu, Haojie Zhang, Xian Yang, Xiaomei Wu, Wei Qi. Relationship between the mRNA transcription levels of AcrAB-TolC efflux pump in Shigella from clinical source and fluoroquinolone resistance[J]. Chinese Journal of Experimental and Clinical Infectious Diseases(Electronic Edition), 2017, 11(06): 556-560.

目的

探讨志贺菌临床分离菌株AcrAB-TolC外排泵的3个组成基因(acrA、acrB、tolC)mRNA转录水平与其氟喹诺酮耐药水平。

方法

收集2009至2015年于天津医科大学总医院、天津医科大学第二医院和天津市儿童医院3家三甲医院就诊腹泻患者的非重复粪便样本,分离培养获得192株志贺菌临床分离菌株,采用K-B纸片法对所有菌株进行体外药敏试验;根据药敏试验结果,将志贺菌株分为氟喹诺酮耐药组和氟喹诺酮全敏感组;应用实时荧光定量PCR(RT-PCR)技术,检测上述两组志贺菌临床分离株AcrAB-TolC外排泵基因在mRNA转录水平的相对表达量,使用SPSS 17.0软件对结果进行统计学分析。

结果

氟喹诺酮耐药组志贺菌株共计9株(对3种及3种以上氟喹诺酮耐药的菌株),依次编码为F2、N8、F44、157、187、368、1113、3171和3327,氟喹诺酮全敏感组志贺菌株共计9株,依次编码为F13、1、22、174、186、377、1506、3283和3326;耐药组外排泵acrA基因的mRNA转录水平高于全敏感组菌株,差异具有统计学意义(t = 2.97、P = 0.02);耐药组外排泵acrB和tolC基因的mRNA转录水平均值虽然高于全敏感组相应基因mRNA转录水平,但差异均无统计学意义(t = 2.84、P = 0.30,t = 1.17、P = 0.29)。

结论

受试菌株外排泵acrA基因mRNA转录水平增高与氟喹诺酮耐药存在一定的关联。

Objective

To investigate the relationship between the transcriptional levels of messenger RNA (mRNA) from component genes (acrA, acrB, tolC) of the efflux pump AcrAB-TolC of clinical isolates of Shigella flexneri and the level of fluoroquinolone resistance.

Methods

Total of 192 clinical Shigella isolates were collected from non-repetitive faeces of diarrhea patients during 2009 to 2015 from 3 grand three hospitals in Tianjin, which were The General Hospital of Tianjin Medical Univercity, The Second Hospital of Tianjin Medical Univercity and The Children’s Hospital in Tianjin. Antibiotic susceptibility of the Shigella clinical isolates were detected by K-B disk diffusion. The strains were divided into the fluoroquinolone resistant group and full fluoroquinolone sensitive group, according to susceptibility to fluoroquinolone. The mRNA transcription of AcrAB-TolC efflux pump of the two groups of strains determined by real-time quantitative PCR (RT-PCR). And the SPSS 17.0 software was used for statistical analysis.

Results

There were 9 isolates in fluoroquinolone resistance group, which were resistant to three and above kinds of fluoroquinolone. And the number in the fluoroquinolone resistance group were F2, N8, F44, 157, 187, 368, 1113, 3171 and 3327. While there were 9 isolates in fluoroquinolone full-sensitive group. And the number were F13, 1, 22, 174, 186, 377, 1506, 3283 and 3326 . The mRNA transcription level of acrA in the resistant group was higher than that of the full-sensitive group, with significant difference (t = 2.97, P = 0.02). Although the mRNA transcription levels of acrB and tolC in the resistant group were higher than those of the full-sensitive group, but with no significant difference (t = 2.84, P = 0.30; t = 1.17, P = 0.29).

Conclusions

There was a certain correlation between the high mRNA transcription level of acrA and the resistance of fluoroquinolones in Shigella.

表1 实时荧光定量PCR反应引物一般特征
表2 耐药组和全敏感组菌株的筛选结果
表3 氟喹诺酮耐药组实验菌株3个目的基因的相对表达量
表4 AcrAB-TolC外排泵mRNA相对表达水平( ± s
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