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猪链球菌14型的分离鉴定及生物学特性研究
引用本文:王治方,徐引弟,张青娴,朱文豪,白红杰,焦文强,李海利,许峰,王克领,张彬,娄治国.猪链球菌14型的分离鉴定及生物学特性研究[J].中国畜牧兽医,2020,47(6):1861-1869.
作者姓名:王治方  徐引弟  张青娴  朱文豪  白红杰  焦文强  李海利  许峰  王克领  张彬  娄治国
作者单位:1. 河南省农业科学院畜牧兽医研究所, 郑州 450002;2. 河南省畜禽繁育与营养调控重点实验室, 郑州 450002;3. 河南省农业科学院, 郑州 450002
基金项目:河南省农业科学院自主创新基金项目(2019ZC39)
摘    要:为了检测确定2019年5月河南某规模化猪场一栋保育仔猪发病猪群的病原,本研究从送检的发病猪关节液中分离获得1株细菌。通过细菌纯化培养、革兰氏染色、形态学观察及猪链球菌gdh基因PCR扩增,确定该分离菌株为猪链球菌。用猪链球菌分型引物对该菌株进行PCR扩增分型鉴定及软件比对分析,结果表明该分离菌株为猪链球菌14型,与猪链球菌JS14株(GenBank登录号:CP002465.1)同源性为100%。毒力基因检测结果表明,该菌株的同时携带有epf、mrp、sly、fbps、orf2毒力基因,属于高致病性菌株。小鼠致病性试验结果也证明该菌株是一株高致病性猪链球菌。药物敏感性试验结果显示,该菌株对β内酰胺类和喹诺酮类药物敏感,对氨基糖苷类、四环素类、大环内酯类和磺胺类高度耐药,表现出多重耐药现象。对该菌株进行5大类24种耐药基因检测,该菌株同时携带有blaTEMaadA1、strA、strB、aacC2、aphA1、tet(B)、gyrA、parC、sul2耐药基因。该研究为后续进一步开展猪链球菌14型流行特点和致病机制研究奠定了基础,为猪链球菌14型临床防控提供了理论依据,同时具有重要的公共卫生意义。

关 键 词:链球菌14型  分型鉴定  毒力基因  药敏试验  耐药基因    
收稿时间:2020-01-10

Isolation,Identification and Biological Characteristics of Streptococcus suis Serotype 14
WANG Zhifang,XU Yindi,ZHANG Qingxian,ZHU Wenhao,BAI Hongjie,JIAO Wenqiang,LI Haili,XU Feng,WANG Keling,ZHANG Bin,LOU Zhiguo.Isolation,Identification and Biological Characteristics of Streptococcus suis Serotype 14[J].China Animal Husbandry & Veterinary Medicine,2020,47(6):1861-1869.
Authors:WANG Zhifang  XU Yindi  ZHANG Qingxian  ZHU Wenhao  BAI Hongjie  JIAO Wenqiang  LI Haili  XU Feng  WANG Keling  ZHANG Bin  LOU Zhiguo
Institution:1. Institute of Animal Husbandry and Veterinary Research, Henan Academy of Agricultural Sciences, Zhengzhou 450002, China;2. Henan Key Laboratory of Farm Animal Breeding and Nutritional Regulation, Zhengzhou 450002, China;3. Henan Academy of Agricultural Sciences, Zhengzhou 450002, China
Abstract:In order to detect and determine the pathogen of the diseased piglets in a large-scale pig farm in Henan province in May 2019,a strain of bacteria was isolated from the joint fluid of diseased piglets.The isolated strain was identified as Streptococcus suis by purification culture,Gram staining,morphological observation and PCR amplification of gdh gene of Streptococcus suis.PCR amplification and software were used to identify the Streptococcus suis.The results showed that the strain was Streptococcus suis serotype 14 and 100% homologous with js14 (GenBank No.:CP002465.1).The results of virulence gene detection showed that epf,mrp,sly,fbps and orf2 virulence genes were carried by the strain at the same time.The results of mouse pathogenicity test also showed that the strain was a highly pathogenic Streptococcus suis.The results of drug sensitivity test showed that the strain was sensitive to β-lactams and quinolones,and highly resistant to aminoglycosides,tetracyclines,macrolides and sulfonamides,showing multiple drug resistance.The resistance genes of blaTEM,aadA1,strA,strB,aacC2,aphA1,tet(B),gyrA,parC and sul2 were detected.This study laid a foundation for further research on the epidemic characteristics and pathogenic mechanism of Streptococcus suis serotype 14,provided theoretical basis for clinical prevention and control of Streptococcus suis serotype 14,and had important public health significance.
Keywords:Streptococcus suis serotype 14  typing and identification  virulence gene  drug sensitivity test  drug resistance gene  pigs  
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