兔支气管败血波氏杆菌的分离鉴定及耐药基因检测

Isolation,Identification and Detection of Drug Resistant Genes of Rabbit Bordetella bronchiseptica

本试验旨在了解近年来浙江省兔支气管败血波氏杆菌的感染情况及耐药状况,指导合理用药,检测细菌的耐药基因,探究其耐药机理。根据细菌形态、培养特性、生化试验,结合PCR法对分离菌株进行鉴定;K-B纸片扩散法检测细菌对18种常用抗生素的耐药率;设计耐药基因特异性引物,PCR法扩增耐药基因,并进行测序分析。结果显示,分离鉴定出2012—2014年22株兔支气管败血波氏杆菌;药敏结果显示,分离菌对青霉素G、头孢拉定、链霉素、林可霉素、克林霉素、甲氧嘧啶耐药严重,对多黏菌素B、左氟沙星、强力霉素、四环素等药物敏感,耐β-内酰胺类药物的细菌中检测到耐药基因blaTEM。结果表明,兔支气管败血波氏杆菌是引起兔呼吸道疾病的主要病原菌,分离菌多重耐药,耐药率较高,检测到的耐药基因与耐药表型相符。

In order to know the infection and drug resistance of Bordetella bronchiseptica from rabbit in Zhejiang province in recent years and guide the rational use of drugs,resistance genes were detected to study the possible mechanism of drug resistance.The bacteria were identified via the test of mophology,culture distinguishing and biochemical test combined with PCR method;K-B＇s paper method was used to detect the drug resistance rate for 18 kinds of common antibiotics;Specific primers were also designed to detect through PCR method.Twenty-two Bordetella bronchiseptica strains were isolated and identified during 2012-2014.Drug susceptibility results showed that bacteria were highly resistant to penicillin G,cefradine,streptomycin,lincomycin,clindamycin and sulfamnomethoxine,and sensitive to polymyxin,levofloxacin,doxycycline and tetracycline,etc;blaTEM gene from the bacteria was detected positive which were resistant to beta-lactams.Rabbit respiratory disease was mainly caused by Bordetella bronchiseptica,the isolated bacteria were multi-drug resistance,drug resistance rate was high,the resistant genes were consistant with drug-resistant phenotype.