毛皮动物源肺炎克雷伯菌部分毒力基因、耐药基因检测及药敏试验

Detection of virulence gene and resistance gene and drug sensitivity test of Klebsiella Pneumoniae in fur -bearing animal

为了了解河北秦皇岛毛皮动物(狐、貉、貂)源肺炎克雷伯菌毒力基因、耐药基因分布及药物敏感性等情况,采用常规分离纯化、生化鉴定及分子生物学的方法对病原菌进行鉴定,动物致病性试验检测其致病性,PCR方法检测其主要K血清型、毒力基因和耐药基因,K-B法对常用西药进行药物敏感性检测,琼脂平板打孔法对中药药物敏感性进行检测,试管二倍稀释法检测中药最小抑菌浓度(MIC)和最小杀菌浓度(MBC)。结果显示:此次共分离到48株肺炎克雷伯菌;48株肺炎克雷伯菌均能够使小鼠发病死亡;48株肺炎克雷伯菌均检测到fimH、wabG毒力基因,检出率为100%;mrkD毒力基因检出率为87.5%;blaTEM、aadA1、gyrB、gyrA、blaSHV-1、sul1、sul2、sul3、blaROB-1、tetA和tetD耐药基因的检出率分别为100.0%、100.0%、89.6%、4.2%、39.6%、18.8%、27.1%、16.7%、12.5%、60.4%和37.5%,未检出ermF耐药基因。西药药敏试验结果显示,48株肺炎克雷伯菌临床分离菌株均对头孢拉定、复方新诺明、林可霉素、替米考星、磺胺间甲氧嘧啶耐药。中药药敏试验结果显示,48株肺炎克雷伯菌临床分离菌株对乌梅、五味子极度敏感,抑菌圈直径在20~25 mm之间,MIC和MBC为15.63~62.50 g/L。本试验调查了河北秦皇岛地区毛皮动物源肺炎克雷伯菌的流行情况,对毛皮动物源出血性肺炎的防治具有重要的指导意义,更具公共卫生学意义。

In order to understand the distribution of virulence genes and drug resistance genes and drug sensitivity of Klebsiella pneumoniae in fur-bearing animal(fox,raccoon dog,mink)from Qinhuangdao City,Hebei Province,biological methods of routine isolation,purification,biochemical identification,and molecular analysis were used to identify pathogenic bacteria,its pathogenicity was detected by animal pathogenicity test,the major K serotypes,virulence genes and drug resistance genes were detected by PCR,the sensitivity of commonly used western drugs were detected by K-B methods;the sensitivity of Chinese herbal medicines were detected by agar plate drilling method,and the minimum inhibitory concentrations(MICs)and the minimum bactericidal concentrations(MBCs)of Chinese herbal medicines were detected by tube double dilution method.The test results showed that 48 strains of Klebsiella pneumoniae were isolated in this time and could cause 100%death of mice after intraperitoneal injection;the detection rates of virulence genes of fimH,wabG were detected in 48 strains of Klebsiella pneumonia were 100%;the detection rate of mrkD was 87.5%;that of blaTEM,aadA1,gyrB,gyrA,blaSHV-1,sul1,sul2,sul3,blaROB-1,tetA and tetD were 100.0%,100.0%,89.6%,4.2%,39.6%,18.8%,27.1%,16.7%,12.5%,60.4%and 37.5%,respectively;However,the resistance gene of ermF was not detected;Western medicine susceptibility test results showed that 48 strains were resistant to cefradine,cotrimoxazole,lincomycin,tilmicosin,sulfamonomethoxine.Chinese herbal medicines susceptibility test results showed that Prunus mume,Schisandra chinensis had better in vitro antibacterial effect on48 clinical strains of Klebsiella pneumoniae against,the inhibition zone diameter were from 20 to25 mm,and MIC and MBC were from 15.63-62.5 g/L.This study investigated the prevalence of Klebsiellapneumoniaein fur-bearing animal in Qinhuangdao,Hebei Province,and had important guiding significance for the prevention and treatment of hemorrhagic pneumonia in fur-bearing animal and public health significance.