中国部分地区猪源和牛源金黄色葡萄球菌耐药性及凝固酶分型研究

doi:10.3864/j.issn.0578-1752.2012.17.018

摘要/Abstract

摘要： 【目的】了解中国动物源金黄色葡萄球菌的耐药现状，以及凝固酶血清型和基因型分布情况，为控制动物源金黄色葡萄球菌的传播和流行提供理论依据。【方法】用微量肉汤稀释法测定细菌耐药性；用凝固酶分型血清确定血清型别；扩增凝固酶基因（coa）并进行序列测定，用mega软件进行序列分析。【结果】共鉴定出124株金黄色葡萄球菌。耐药性检测结果表明，在13种测试抗菌药中，金黄色葡萄球菌对氨苄西林和青霉素的耐药率最高，接近100%，其次是红霉素75%，其它10种药物的耐药率在50%以下，头孢西丁耐药率27%，未发现万古霉素耐药菌株；凝固酶血清分型率达91%，共分出6个型，VII型，VI型和复合型占优势，未分离到Ⅲ型和Ⅴ型；扩增凝固酶基因，共分为6个基因型；构建了凝固酶基因的系统进化树。【结论】中国动物源金黄色葡萄球菌的耐药状况比较严重，猪源菌株的耐药率要高于牛源菌株。凝固酶血清型别多样，分布相对较集中，具有独特的流行特征；凝固酶表型与其基因的进化特征并没有出现直接对应关系，但为进一步开展兽医临床金黄色葡萄球菌分子流行病学调查研究奠定了理论基础。

关键词: 金黄色葡萄球菌, 耐药性, 凝固酶分型, 猪, 牛

Abstract: 【Objective】The objective of the experiment is to study the status of antibiotics resistance and coagulase typing distribution of S.aureus in China, and provide a scientific basis for effective control of transmission and prevalence of S.aureus from animals.【Method】Antibiotics resistance was tested by broth dilution method, coagulase serotypes were identified by coagulase grouping serum, coa genes were amplified by PCR method and then sequenced for further analysis by mega software. 【Result】 A total of 124 S.aureus isolates were identified, and the result of resistance detection revealed that the resistant rate of ampicillin and penicillin was near 100%, erythromycin reached 75%, and other 10 antibiotics was below 50%. The positive rate of MRSA was 27%. No vancomycin resistant isolates was found. Six kinds of serotypes were observed and the rate of coagulase serotyping reached 91%: serotypes VII,VI and combined type were prevalent, whereas serotypes Ⅲ and Ⅴwere not found. The phylogenetic tree of coagulase gene was constructed. 【Conclusion】 The status of antibiotics resistance in China was relatively severe, swine isolates were severer than cattle ones. Coagulase serotypes were abundant and the distribution was comparative intensive, and possessed unique prevalence feature. The evolutional characteristic of coa gene had no direct relationship with coagulase serotypes, and this study has provided a theoretical basis for further veterinary research about molecular epidemiology and evolutionary tendency.

Key words: S. aureus, antibiotics resistance, coagulase typing, pig, cow

刘洋, 梁耀峰, 焦新安, 宋立, 张纯萍, 宁宜宝, 徐士新. 中国部分地区猪源和牛源金黄色葡萄球菌耐药性及凝固酶分型研究[J]. 中国农业科学, 2012, 45(17): 3608-3616.

LIU Yang, LIANG Yao-Feng, JIAO Xin-An, SONG Li, ZHANG Chun-Ping, NING Yi-Bao, XU Shi-Xin. Antibiotic Resistance and Coagulase Typing of Staphylococcus aureus Isolates from Pigs and Cows in Part of China[J]. Scientia Agricultura Sinica, 2012, 45(17): 3608-3616.

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参考文献

[1]陆承平. 兽医微生物学. 北京: 中国农业出版社, 2006: 200-204.

Lu C P. Veterinary Microbiology. Beijing: China Agriculture Press, 2006: 200-204. (in Chinses)

[2]Le Hoir Y, Baron F, Gautier M. Staphylococcus aureus and food poisoning. Genetic Molecular Research, 2003, 2: 63-76.

[3]Martin M C, Fueyo J M, Gonzalez M A. Genetic procedures for identification of enterotoxigenic strains of staphylococcus aureus from three food poisoning outbreaks. International Journal of Food Microbiology, 2004, 94:279-286.

[4]王英杰, 王长法, 陈营, 安利国, 杨桂文, 仲跻峰, 杨少华, 杨宏军. PCR扩增血浆凝固酶基因检测致病性金黄色葡萄球菌. 生物技术通报, 2008, 5:185-188.

Wang Y J, Wang C F, Chen Y, An L G, Yang J W, Zhong J F, Yang S H, Yang H J. Identification of coagulase gene of pathogenic Staphylococcus aureus bovine mastitis by PCR. Biotechnology Bulletin, 2008, 5:185-188.(in Chinese)

[5]代文霞, 沈叙庄, 杨永弘. 金黄色葡萄球菌的流行病学分型方法研究进展. 国外医学儿科学分册, 2004, 31(5):240-242.

Dai W X, Shen X Z, Yang Y H. Research progression of epidemiological typing method of staphylococcus aureus. Foreign Medical Sciences Sectin of Pediatrics, 2004, 31(5):240-242. (in Chinese)

[6]倪春霞, 蒲万霞, 胡永浩, 王玲, 邓海平. 金黄色葡萄球菌基因分型方法研究进展. 中国动物检疫, 2009, 26(10):64-68.

Ni C X, Pu W X, Hu Y H, Wang L, Deng H P. Research progression of genotyping method of staphylococcus aureus. China Animal Quarantine, 2009, 26(10):64-68. (in Chinese)

[7]朱战波, 任宪刚, 崔玉东, 朴范泽. 牛源金黄色葡萄球菌凝固酶基因的分型. 中国兽医学报, 2007,27(5):728-730.

Zhu Z B, Ren X G,, Cui Y D, Piao F Z. Coagulase gene typing of Staphylococcus aureus isolated from bovine. Chinese Journal of Veterinary Science, 2007, 27(5): 728-730.(in Chinese)

[8]许一平, 成炜, 邵彦春, 陈福生. 沙门菌、大肠杆菌和金黄色葡萄球菌的多重PCR检测. 微生物学通报, 2006, 3(6):89-94.

Xu Y P, Cheng W, Shao Y C, Chen F S. Detection of Salmonella spp1, Escherichia coli and Staphylococcus aureus by Multiplex PCR. Journal of Microbiology, 2006, 3(6):89-94. (in Chinese)

[9]Clinical and Laboratory Standards Institute. Performance standards for antimicrobial susceptibility testing; eighteenth informational supplement, M100-S19[S]. Wayne, PA, 2009.

[10]倪春霞, 蒲万霞, 胡永浩, 邓海平. 奶牛乳房炎金黄色葡萄球菌凝固酶基因型研究. 中国农业科学, 2011, 44(2): 417-422.

Ning C X, Pu W X, Hu Y H, Deng H P. Research on coagulase genotyping of Staphylococcus aureus isolated from bovine mastitis. Scientia Agricultura Sinica, 2011, 44(2):417-422. (in Chinese)

[11]任宪刚, 朱战波, 崔莉, 宋佰芬, 谭磊, 崔玉东, 朴范泽. 不同凝固酶基因型牛源金黄色葡萄球菌主要致病因子的检测及分析. 中国预防兽医学报, 2008, 30(8):608-611.

Ren X G, Zhu Z B, Cui L, Song B F, Tan L, Cui Y D, Piao F Z. Virulence factors of Staphylococcus aureus of different coagulase genotype. Chinese Journal of Preventive Veterinary Medicine, 2008, 30(8):608-611.(in Chinese)

[12]Kumar S, Tamura K, Nei M. MEGA3: Integrated software for molecular evolutionary genetics analysis and sequence alignment. Briefings in Bioinformatics, 2004, 5:150-163.

[13]Thompson J D, Higgins D G, Gibson T J. CLUSTAL W: Improving the sensitivity of progressive multiple sequence alignment through sequence weighting, positions-specific gap penalties and weight matrix choice. Nucleic Acids Research, 1994, 22:4673-4680.

[14]王辉, 孙宏莉, 陈民钧, 胡必杰, 俞云松, 孙自庸, 褚云卓, 徐英春, 谢秀丽. 2005年我国五家教学医院革兰阳性球菌耐药监测研究. 中华检验医学杂志, 2006, 29: 873-877.

Wang H, Sun H L, Chen M J, Hu B J, Yu Y S, Sun Z Y, Chu Y Z, Xu Y C, Xie X L. Antimicrobial resistance surveillance of gram-positive cocci isolated from 5 teaching hospitals in China in 2005. Chinese Journal of Labrotory Medicine, 2006, 29:873-877. (in Chinese)

[15]万钧, 李羲, 贾坤茹, 陈美华, 胡龙华. 医院与社区获得性耐甲氧西林金黄色葡萄球菌耐药性的研究. 中华医院感染学志, 2005, 15(6):668-670.

Wan J, Li Y, Jia K R, Chen M H, Hu L H. Drug resistance of meticillin-resistant Staphylococcus aureus. Chinese Journal of Nosocomoil, 2005, 15(6):668-670. (in Chinese)

[16]王桂琴, 杨萌萌, 邢燕, 尹力媛, 刘伟, 冯若洋. 宁夏地区奶牛乳房炎金黄色葡萄球菌耐药性分析. 动物医学进展， 2011, 32(10): 59-62.

Wang G Q, Yang M M, Xing Y, Liu W, Feng R Y. Drug resistance analysis of Staphylococcus aureus from dairy cow mastitis in Ningxia area. Progess in Veterinary Medicine, 2011, 32(10):59-62.(in Chinese)

[17]邱梅, 郝智慧, 张万江, 沈巍, 吴彪, 李玉华, 张桂贤. 动物源金黄色葡萄球菌的分离鉴定及其耐药性分析. 中国农学通报, 2011, 27(7): 356-359.

Qiu M, Hao Z H, Zhang W J, Shen W, Wu B, Li Y H, Zhang G X. Isolation and Identification of Staphylococcus aureus from Animals and Drug Resistance Analysis. Chinese Agricultural Science Bulletin, 2011, 27(7): 356-359. (in Chinese)

[18]孙怡群, 徐修礼, 杨秋红, 樊新, 彭道荣. 金黄色葡萄球菌凝固酶分型及耐药性研究分析. 临床检验杂志, 2004, 22(4): 293-294.

Sun Y Q, Xu X L, Yang Q H, Fan X, Peng D R. Research on antibiotic resistance and coagulase typing of Staphylococcus aureus. Chinese Journal of Clinical Laboratory Science, 2004, 22(4): 93-294. (in Chinese)

[19]Shinya W, Teruyo I, Takashi S, Shanshuang L, Ikuo U, Kozue K, Ken K, Robert L, Keiichi H. Genetic diversity of Staphylocoagulase Genes (coa): Insight into the evolution of variable chromosomal virulence factors in Staphylococcus aureus. PLoS ONE, 2009, 4(5): e5714.

[20]Berglund C, Ito T, Ma XX, Ikeda M, Watanabe S, Soderquist B, Hiramatsu K. Genetic diversity of methicillin-resistant Staphylococcus aureus carrying type IV SCCmec in orebro county and the western region of Sweden. Journal of Antimicrobial Chemotherapy, 2009, 63: 32–41.

[21]谭磊. 基于PCR的牛源金黄色葡萄球菌基因分型研究[D]. 大庆：黑龙江八一农垦大学, 2008.

Tan L. The Study on PCR-based genotyping of bovine source staphylococcus aureus[D]. Daqing: Heilongjiang August First Land Agricultural University, 2008.(in Chinese)

[22]Güler L, Ok U, Gündüz K, GülcüY, Hadimli H H. Antimicrobial susceptibility and coagulase gene typing of Staphylococcus aureus isolated from bovine Clinical Mastitis Cases in Turkey. Journal of Dairy Science, 2005, 88(9):3149-3154.

[23]Montesinos I, Salido E, Delgado T, Cuervo M, Sierra A. Epidemiologic genotyping of methicillin-resistant Staphylococcus aureus by pulsed-field gel electrophoresis at a university hospital and comparison with antibiotyping and protein A and coagulase gene polymorphisms. Journal of Clinical Microbiology, 2002, 40(6): 2119-2125.

[24]Kim Y K, Kim, J S. Kim H S, Song W, Cho H C, Lee K M. Molecular typing of Staphylococcus aureus isolated from blood on the basis of coagulase gene polymorphism and toxin genes. The Korean Journal of Laboratory Medicine, 2008, 28(4):286-292.

[25]Tiwari, H. K, Sapkota, D, Gaur, A, Mathuria J P, Singh A, Sen M R. Molecular typing of clinical Staphylococcus aureus isolates from northern India using coagulase gene PCR-RFLP. Southeast Asian Journal of Tropical Medicine and Public Health ,2008, 39(3):467-473.

[26]Saei H D, Ahmadi M, Mardani K, Batavani R A. Molecular typing of Staphylococcus aureus isolated from bovine mastitis based on polymorphism of the coagulase gene in the north west of Iran. Veterinary Microbiology, 2009, 137(1,2):202-206.

[27]Michiru K, Yuichi H, Teisuya O, Hirotaka K, Kazuhiro T, Hajime K, Fumiko K. Ribotyping and a study of transmission of Staphylococcus aureus collected from food preparation facilitiess. Journal of Food Protection, 2004, 67(6):1116–1122.

[28]Watanabe S, Ito T, Takeuchi F, Endo M, Okuno E, Hiramatsu K. Structural comparison of ten serotypes of staphylocoagulases in Staphylococcus aureus. Journal of Bacteriology, 2005, 187(11) 3698-3707.