| 云南地区牛源无乳链球菌分离鉴定及毒力基因和耐药性的检测 |
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| 引用本文： | 祝宇,路璐,颜兴琼,龚蕾,孙卫星,曲伟杰.云南地区牛源无乳链球菌分离鉴定及毒力基因和耐药性的检测[J].中国畜牧兽医,2018,45(11):3261-3270. |
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| 作者姓名： | 祝宇 路璐 颜兴琼 龚蕾 孙卫星 曲伟杰 |
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| 作者单位： | 云南农业大学动物医学院, 昆明 650201 |
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| 基金项目： | 国家自然科学基金（3166130435）；云南省高校兽医公共卫生重点实验室 |
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| 摘 要： | 试验旨在研究无乳链球菌的生物学特性，为防治无乳链球菌引起的奶牛乳房炎提供理论依据。根据细菌分子生物学分离鉴定无乳链球菌，参考GenBank登录的牛源无乳链球菌16S rRNA、菌属特异性cfb （CAMP因子）、毒力基因和耐药基因序列，运用Oligo 6.0和Primer Premier 5.0软件设计14对引物，建立PCR快速检测方法，并进行20种常见抗生药物的耐药试验。结果显示，试验成功鉴定出17株牛源无乳链球菌，毒力基因与NCBI上已报道的无乳链球菌相应序列具有高度同源性，均≥99%；共检测到6种耐药基因；分离菌株对青霉素、红霉素、林可霉素、克林霉素、万古霉素、氨苄西林、新生霉素、磺胺异噁唑的耐药率均较高，耐药率依次为100.0%、94.1%、94.1%、94.1%、94.1%、82.3%、82.3%和47.1%，对青霉素严重耐药；而对氨基糖苷类、四环素类、头孢菌素类、喹诺酮类耐药率均较低，耐药率分别为15.7%、14.7%、7.7%和3.9%。本研究结果表明，建立的PCR快速检测方法灵敏可靠，云南地区无乳链球菌已对部分β-内酰胺类、大环内酯类、磺胺类等抗生素出现多重耐药性。
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| 关 键 词： | 奶牛乳房炎 无乳链球菌 毒力基因 耐药性 |
| 收稿时间： | 2018-04-02 |
Isolation,Identification and Detection of Virulence Genes and Drug Resistance of Streptococcus agalactiae from Cattle in Yunnan |
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| ZHU Yu,LU Lu,YAN Xingqiong,GONG Lei,SUN Weixing,QU Weijie.Isolation,Identification and Detection of Virulence Genes and Drug Resistance of Streptococcus agalactiae from Cattle in Yunnan[J].China Animal Husbandry & Veterinary Medicine,2018,45(11):3261-3270. |
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| Authors: | ZHU Yu LU Lu YAN Xingqiong GONG Lei SUN Weixing QU Weijie |
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| Institution: | College of Veterinary Medicine, Yunnan Agricultural University, Kunming 650201, China |
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| Abstract: | This experiment was aimed to study the biological characteristics of Streptococcus agalactiae (S.agalactiae)and provide a theoretical basis for the prevention and treatment of cow mastitis caused by S.agalactiae.S.agalactiae was separated and identified based on bacterial molecular biology;according to GenBank-registered S.agalactiae 16S rRNA,genus-specific cfb (CAMP factor),virulence gene and drug resistance gene sequences,14 pairs of primers were designed using Oligo 6.0 and Primer Premier 5.0 softwares,then,a rapid PCR detection method was established;The resistance tests of 20 common antibiotic drugs was carried out.The results showed that 17 strains of S.agalactiae were successfully identified,and the virulence genes were highly homologous to the corresponding sequences of S.agalactiae reported on NCBI,which were ≥ 99%;A total of 6 drug resistance genes were detected;The resistance rates of the isolates to penicillin,erythromycin,lincomycin,clindamycin,vancomycin,ampicillin,novobiocin and sulfisoxazole were higher,and the drug resistance rates were 100%,94.1%,94.1%,94.1%,94.1%,82.3%,82.3% and 47.1%,respectively,severely resistant to penicillin;And the resistance rates to aminoglycosides,tetracyclines,cephalosporins and quinolones were low,which were 15.7%,14.7%,7.7% and 3.9%,respectively.This study results indicated that the established PCR rapid detection method was sensitive and reliable,and there was a multi-drug resistance of some β-lactams,macrolides,sulfonamides and other antibiotics in Yunnan. |
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| Keywords: | dairy cow mastitis Streptococcus agalactiae virulence gene drug resistance |
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