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为了调查广东省惠州、肇庆、珠海、湛江4个吉富罗非鱼主养区链球菌病的流行情况和耐药性,并进一步分析β-内酰胺酶基因与青霉素类药物的耐药性关系。本实验通过传统的方法对菌株进行分离纯化,扩增特异性基因cfb以及16s r DNA对各菌株进行鉴定;采用k-b法测定分离菌株的药物敏感性;通过PCR检测β-内酰胺酶类基因在分离菌株中的分布情况,并用Statistic6.0统计分析各β-内酰胺酶基因与青霉素类药物的耐药关系。实验结果表明,吉富罗非鱼无乳链球菌的阳性率从高到低的顺序为惠州(46.46%)>湛江(43.24%)>肇庆(17.30%)>珠海(4.17%);药敏结果显示各地区无乳链球菌分离株对青霉素(耐药率为94.29%)和磺胺二甲基嘧啶(耐药率为86.40%)普遍耐药,对恩诺沙星最为敏感(耐药率为3.99%);β-内酰胺酶基因分布与细菌耐药性统计结果显示,无乳链球菌基因组中的9个β-内酰胺酶基因在各分离菌株中的分布呈高度多态性,其中SAG0658基因与氨苄青霉素抗性显著相关,提示SAG0658基因在无乳链球菌耐氨苄青霉素过程中发挥主要作用;此外,9个β-内酰胺酶基因与青霉素抗性无相关性,说明其在菌株对青霉素耐药过程中并未发挥明显作用,提示分离菌株对青霉素的耐药性可能依赖其他途径。
In order to investigate the prevalence and drug resistance of streptococcal disease in four main areas of GIFT tilapia in Guangdong, Huizhou, Zhaoqing, Zhuhai and Zhanjiang, and to further analyze the relationship between β-lactamase gene and penicillin resistance. In this experiment, the strains were isolated and purified by traditional methods, and the specific genes cfb and 16s r were amplified to identify the strains. The drug sensitivity of the isolates was determined by kb method. The β-lactamase gene The distribution of isolates was analyzed. Statistic6.0 was used to analyze the resistance of each β-lactamase gene to penicillin. The experimental results showed that the order of the positive rates of S.juronicus from Streptococcus agalactiae was Huizhou (46.46%), Zhanjiang (43.24%), Zhaoqing (17.30%), Zhuhai (4.17%), Streptococcus agalactiae isolates from all regions were generally resistant to penicillin (resistance rate 94.29%) and sulfamethazine (resistance rate 86.40%), and were most sensitive to enrofloxacin (resistance rate was 3.99% ); β-lactamase gene distribution and bacterial resistance statistics show that the genome of Streptococcus agalactiae 9 β-lactamase gene distribution in each isolates was highly polymorphic, including SAG0658 gene and Ampicillin resistance was significantly correlated, suggesting that SAG0658 gene play a major role in the process of Streptococcus agalactiae resistance to ampicillin; In addition, nine β-lactamase gene and penicillin resistance no correlation, indicating that the strain resistant to penicillin The process did not play a significant role, suggesting that isolates resistant to penicillin may depend on other ways.