多重PCR法检测与分析鱼塘生态系统大肠杆菌的耐药基因与整合子
Detection and analysis of antibiotic resistance genes and integron of Escherichia coli isolated from fish pond ecological constellation by mutiplex PCR
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摘要: 了解在鱼塘生态系统中,大肠杆菌耐药基因、整合酶基因的携带情况,并对其进行分析,可以为控制细菌耐药基因传播提供科学的依据。本文对来自鱼塘生态系统的165株大肠杆菌进行了检测,发现大肠杆菌中普遍带有耐药基因,携带有耐药基因的比率为91.51%。携带有2种耐药基因以上菌株的比率为86.06%,大肠杆菌耐药基因的携带以多重耐药基因为主。进而对带有4种以上耐药基因的菌株检测,Ⅰ整合子酶基因阳性检出率为87.84%,携带有7种耐药基因的菌株全部为Ⅰ整合酶基因阳性;Ⅱ型整合酶基因阳性菌株检出率10.81%;1株为阴性;检测中没发现有携带Ⅲ型整合酶基因菌株。可见Ⅰ整合子介导的耐药机制占主导位置,并且通过整合子的整合作用向周围环境扩散。Abstract: Integrase and integron in E. coli could provide reliable scientific foundation to the control of the spread of antibiotic resistance of bacteria. In this study, assessed the situation of antibiotic resistance genes (ARG) , 165 strains of E. coli isolated from ecological constellation were screened for 19 antibiotic resistance genes using multiplex PCR assays and it was found that 91. 51% of E. coli carried ARGs. The results showed that it was popular to carry ARGs in E. coli strains. Among them, E. coli strains carried multiplex ARGs played a dominant role, and there were about 86. 06% of E. coli strains carried more than 2 ARGs. Then multiplex PCR assay was used to detect the strains carried more than 4 ARGs for integrase genes. The PCR result revealed that the frequency of class I integrase gene was 87. 84%, the proportion of class II integrase gene was 10. 81%. None of class III integrase gene was found in the test. Moreover, the results indicated that integron-cassette system predominated in the antibiotic resistant machanism mediated by integron.
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