四川省一些实验动物养殖场SPF大鼠和小鼠肠内容物中金黄色葡萄球菌的检测
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摘要
调查四川省一些实验动物养殖场的实验动物中金黄色葡萄球菌(Staphylococcus aureus,SA)携带情况,找出传播途径和进化规律,分析其污染原因,为保障实验动物质量提供技术支撑。按照GB/T 14926.14-2001《实验动物金黄色葡萄球菌检测方法》对2011年-2017年四川省部分实验动物中SA进行检测,对分离的SA菌株进行PCR耐药基因mecA检测,并应用金黄色葡萄球菌A蛋白(Staphylococcus protein A,SPA)和脉冲场电泳(pulsed field gel electrophoresis,PFGE)进行分子分型。结果显示,2011年-2017年共从7家实验动物养殖场采集并完成370只实验动物中SA的检测,共检出SA 31株,总分离率为12.16%,且均从SPF大鼠中分离所得。31株SA的mecA基因均为阴性。SPA可分成5个型别,其中T2360为优势型别,PFGE型别有10个带型。四川省养殖实验动物中仅SPF大鼠携带SA,不同年份检出SA基因型别基本不同,不同养殖场同一年检出的SA具有相同基因型,说明不同养殖场中的SA可能来自同一污染源,应加强SPF大鼠监控,以确保实验动物的质量。
To investigate the carrying situation of Staphylococcusaureus(SA)in some laboratory animals in Sichuan province,to find out the way of transmission and the law of evolution,to analyze the cause of contamination,and to provide technical support for ensuring the quality of laboratory animals,according to the National Standard(GB/T 14926.14-2001),the strains of SA in some laboratory animals in Sichuan province in 2011-2017 were isolated and identified.All isolated SA strains were detected for drug-resistance gene mecA by PCR,and the molecular types were carried out by Staphylococcus aureus protein A(SPA)and pulse field electrophoresis(PFGE).31 strains of SA were isolated from 370 samples of laboratory animals,which were collected from seven experimental animal farms in 2011-2017.The total isolation rate was 12.16%.The mecA gene of 31 strains was negative.The SPA typing can be divided into 5 types,of which T2360 is the dominant type,and the PFGE typing can be divided into 10 types.Only SPF rats in the experimental animals of Sichuan province carried SA.The SA genotypes were basically different in different years.The SA in the same year in different farms had the same genotype,indicating that SA in different farms may come from the same source of pollution,the monitoring of SPF rats should be strengthened to ensure the quality of experimental animals.
To investigate the carrying situation of Staphylococcusaureus(SA)in some laboratory animals in Sichuan province,to find out the way of transmission and the law of evolution,to analyze the cause of contamination,and to provide technical support for ensuring the quality of laboratory animals,according to the National Standard(GB/T 14926.14-2001),the strains of SA in some laboratory animals in Sichuan province in 2011-2017 were isolated and identified.All isolated SA strains were detected for drug-resistance gene mecA by PCR,and the molecular types were carried out by Staphylococcus aureus protein A(SPA)and pulse field electrophoresis(PFGE).31 strains of SA were isolated from 370 samples of laboratory animals,which were collected from seven experimental animal farms in 2011-2017.The total isolation rate was 12.16%.The mecA gene of 31 strains was negative.The SPA typing can be divided into 5 types,of which T2360 is the dominant type,and the PFGE typing can be divided into 10 types.Only SPF rats in the experimental animals of Sichuan province carried SA.The SA genotypes were basically different in different years.The SA in the same year in different farms had the same genotype,indicating that SA in different farms may come from the same source of pollution,the monitoring of SPF rats should be strengthened to ensure the quality of experimental animals.
引文
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[2] 萧闵,周艳艳.浅谈实验动物从业人员的继续教育[J].环境与职业医学,2014,31(1):60-62.
[3] 王鹏飞.实验动物金黄色葡萄球菌、绿脓杆菌和肺支原体多重PCR检测方法的建立与初步应用[D].河北石家庄:河北医科大学,2016.
[4] 邹自英,韩黎,熊杰,等.金黄色葡萄球菌临床分离株SPA分型和耐药特征研究[J].中国感染与化疗杂志,2014,14(2):142-145.
[5] 史冬艳,郝永清,张爱荣,等.奶牛金黄色葡萄球菌性乳房炎研究进展[J].动物医学进展,2010,31(7):82-86.
[6] GB/T 14926.14-2001实验动物金黄色葡萄球菌检测方法[S].
[7] Monaco M,Sanchini A,Grundmann H,et al.Aureus typing Group (2010).Vancomycin-heteroresistant phenotype in invasive methicillin-resistant Staphylococcus aureus isolates belonging to spa type 041 [J].Eur J Clin Microbiol Infect Dis,2010,29:771-777.
[8] Mathema B,Mediavilla J,Kreiswirth BN.Sequence analysis of the variable number tandem repeat in Staphylococcus aureus protein A gene:spa typing[J].Meth Mol Biol,2008,431:285-305.
[9] Mulvey M R,Chui L,Ismail J,et al.Canadian Committee for the Standardization of Molecular Methods.Development of a Canadian standardized protocol for subtyping methicillin-resistant Staphylococcus aureus using pulsed-field gel electrophoresis[J].J Clin Microbiol,2001,39(10):3481-3485.
[10] 杨小蓉,黄伟峰,谢晓丽,等.一例由阪崎肠杆菌感染引起腹泻患儿的溯源分析[J].疾病监测,2014,29(10):794-796.
[11] Hayashimoto N,Morita H.Current microbiological status of laboratory mice and rats in experimental facilities in Japan[J].Exp Anim,2013,62(1):41-48.
[12] 葛文平,张旭,高翔,等.我国商业化SPF级小鼠病原体污染分析[J].中国比较医学杂志,2012,22(3):65-68.
[13] 卢杰.牛源金葡菌抗体检测间接ELISA方法的建立与应用[D].黑龙江大庆:黑龙江八一农垦大学,2006.
[14] 谭磊,朱战波,崔玉东,等.牛源金黄色葡萄球菌spa基因多态性分型研究[J].中国人兽共患病学报,2008,24(2):146-149
[15] 孟祥兆,李桂玲,陈勇,等.临床分离金黄色葡萄球菌的流行病学特征分析[J].中国卫生检验杂志,2014(7):1040-1043.
[16] Liu Y,Wang H,Du N,et al.Molecular evidence for spread of two major methicillin-resistant Staphylococcus aureus clones with a unique geographic distribution in Chinese hospitals[J].Antimicrob Agents Chemother,2009,53(2):512-518.
[17] 叶蕊,石丽媛,王鹏,等.脉冲场凝胶电泳技术简介及其在细菌分子分型中的应用[J].中国媒介生物学及控制杂志,2013,24(2):182-185.