Detection of resistance gene NDM and ESBLs of Enterobacteriaceae and study on its sensitivity enhanced by Pulsatilla decoction

摘要:

采用美国临床实验室标准化协会(Clinical and Laboratory Standards Institute,CLSI)推荐的方法,检测了临床分离的44株肠杆菌科细菌产超广谱β-内酰胺酶(Extended-Spectrum Beta-Lactamases,ESBLs)的情况.利用PCR对临床分离菌进行新德里金属β-内酰胺酶(New Delhi metallo-β-lactamase,NDM)以及ESBLs耐药基因的检测.用微量肉汤稀释法测定了分离菌对阿莫西林等14种抗菌药的敏感性,并统计其耐药率.结果表明,临床分离的44株肠杆菌科细菌中,有31株产超广谱β-内酰胺酶,检出率为70.5%.耐药基因检测结果显示,耐药基因NDM-1携带率为13.6%.ESBLs耐药基因TEM,SHV,CTX-M,OXA的携带率分别为100%,43.2%,45.5%,6.8%.同时发现,有6株菌株同时检出了含NDM-1和ESBLs的基因,有9株菌株同时检出含有2种ESBLs基因,3株同时检出含有3种ESBLs基因.敏感性测定结果显示,分离菌大多呈现多重耐药,分离菌除对替加环素(18.2%)和阿米卡星(36.4%)的耐药率较低外,对其他药物呈现出较高的耐药率,其中对阿莫西林的耐药率高达97.7%,对头孢曲松、环丙沙星、土霉素、四环素、红霉素的耐药率均在75.0%以上.此外发现,同时携带多个耐药基因的菌株耐药率较高.中药复方白头翁汤对分离菌表现出了一定的抑菌活性,其MIC值为204.8 g/L.14种抗菌药分别与白头翁汤联用后,耐药率均有不同程度下降,其中头孢类药物、氟喹诺酮类药物与白头翁汤联用后,耐药率下降较为明显,如头孢曲松由84.1%下降至56.8%,环丙沙星由75.0%下降至45.5%.综上,NDM和ESBLs耐药基因在肠杆菌科细菌中已有一定的流行,同时携带NDM和ESBLs多个耐药基因的菌株耐药较为严重.白头翁汤与抗菌药联用能在一定程度增强耐药菌株的敏感性,为临床相关感染的防治提供理论依据.

Extended-Spectrum Beta-Lactamases(ESBLs)-producing from 44 strains of Enterobacteriaceae were detected by using the method recommended by the Clinical and Laboratory Standards Institute(CLSI).New Delhi metallo-β-lactamase(NDM)and ESBLs resistance genes were detected in clinical isolates by PCR.The susceptibility of the isolates to 14 kinds of antibacterial drugs such as amoxicillin was determined by the micro-broth dilution method,and the resistance rate was calculated.The results showed that among the 44 strains of Enterobacteriaceae,31 strains produced ESBLs,and the detection rate was 70.5%.The results of resistance gene detection showed that the carrier rate of resistance gene NDM-1 was 13.6%.The carrier rates of ESBLs resistance genes TEM,SHV,CTX-M and OXA were 100%,43.2%,45.5%and 6.8%,respectively.At the same time,it was found that 6 strains were detected with NDM-1 and ESBLs genes,9 strains were detected with 2 ESBLs genes,and 3 strains were detected with 3 ESBLs genes.The results of susceptibility testing showed that most of the isolates showed multi-drug resistance,and the isolates showed high resistance rates to other drugs except for tigecycline(18.2%)and amikacin(36.4%).Among them,the resistance rate to amoxicillin was as high as 97.7%,and the resistance rate to ceftriaxone,ciprofloxacin,oxytetracycline,tetracycline and erythromycin were all above 75.0%.In addition,it was found that the strains carrying multiple resistance genes at the same time had a higher resistance rate.The traditional Chinese medicine compound Pulsatilla Decoction showed a certain antibacterial activity against the isolated bacteria,and its MIC value was 204.8 g/L.After 14 kinds of antibacterial drugs were used in combination with Pulsatilla Decoction,the drug resistance rates decreased to varying degrees.Among them,cephalosporins and fluoroquinolones were used in combination with Pulsatilla Decoction,and the drug resistance rates decreased significantly.For example,ceftriaxone dropped form 84.1%to 56.8%,and ciprofloxacin dropped from 75.0%to 45.5%.In conclusion,NDM and ESBLs resistance genes have been prevalent in Enterobacteriaceae,and strains carrying multiple NDM and ESBLs resistance genes are more resistant.The combination of Pulsatilla Decoction and antibacterial drugs can enhance the sensitivity of resistant strains to a certain extent,providing a theoretical basis for the prevention and treatment of clinically related infections.

作者:

许二平 刘保光 董颖 白明 谢苗 汪保英 吴华 李永伟

Xu Erping;Liu Baoguang;Dong Ying;Bai Ming;Xie Miao;Wang Baoying;Wu Hua;Li Yongwei(Academy of Chinese Medical SciencesHenan University of Chinese Medicine,Zhengzhou 450046,China;College of Traditional Chinese MedicineHenan University of Chinese Medicine,Zhengzhou 450046,China;The Second Affiliated Hospital,Henan University of Chinese Medicine,Zhengzhou 450046,China;College of Veterinary Medicine,Henan Agricultural University,Zhengzhou 450002,China)

机构地区:

河南中医药大学中医药科学院 河南中医药大学中医学院 河南中医药大学第二附属医院 河南农业大学动物医学院

出处《betway官方app 学报：自然科学版》 CAS 北大核心 2023年第1期106-113,共8页

Journal of Henan Normal University(Natural Science Edition)

基金:

国家自然科学基金(81973739) 河南省中医药科学研究专项课题(2019ZY1019 20-21ZY2144) 河南省自然科学基金(222300420481).

关键词:

肠杆菌科 超广谱Β-内酰胺酶 白头翁汤 药物敏感性

Enterobacteriaceae ESBLs pulsatilla decoction antimicrobial susceptibility

分类号:

R285.5 [医药卫生—中药学]

肠杆菌科细菌耐药基因NDM和ESBLs的检测及白头翁汤增强其敏感性研究.pdf