动物源金黄色葡萄球菌耐药性、oqxAB和fexA基因流行性及spa分型研究
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摘要
为了解河南省不同动物源金黄色葡萄球菌的耐药情况、oqxAB和fexA基因的流行情况,分析spa基因多态性分型特点。用微量肉汤稀释法测定了分离菌对10种抗菌药物的敏感性,利用PCR方法检测金黄色葡萄球菌中的oqxAB和fexA耐药基因,扩增spa基因的X区域进行分型研究。结果表明,130株金黄色葡萄球菌分离株对大多数药物产生耐药,青霉素、红霉素、林可霉素、四环素、环丙沙星、恩诺沙星、氟苯尼考和喹乙醇对分离菌的耐药率分别为94.62%,95.38%,92.31%,70.77%,55.38%,57.69%,32.31%,19.23%,但对替加环素、万古霉素较敏感。PCR检测发现,35株携带fexA基因,检出率为26.92%。17株携带oqxAB基因,检出率为13.08%。此外,有10株同时携带oqxAB和fexA基因,耐药性检测发现这10株菌耐药性较为严重。130株分离菌中107株菌可通过spa分型,分为21种spa型,最常见的spa型是t15075 (15.38%),其次是t189 (13.85%),t034 (9.23%),t091 (7.69%),t127 (6.92%),t164 (6.15%)。此外,只有t034 (9.23%)和t3512 (3.85%)能够出现在3种不同动物源(分别是牛、猪、鸡及牛、猪、鸭)的菌株中。综上,oqxAB和fexA基因在河南省的动物养殖场中已有一定的流行,同时携带oqxAB和fexA基因的菌株耐药性较为严重,spa型别具有多样性和差异性的特点,故临床应加强耐药性及分型监测。
The prevalence of oqxAB and fexA genes and the polymorphisms of spa gene were presented to understand the drug resistance of Staphylococcus aureus from different animal sources in Henan province.The sensitivity of the isolated bacteria to 10 antimicrobial agents was determined by the microbroth dilution method,the oqxAB and fexA resistance genes in Staphylococcus aureus were detected by PCR,the X region of spa was studied.The results showed that 130 strains of Staphylococcus aureus were resistant to most drugs,penicillin,erythromycin,clindamycin,tetracycline,ciprofloxacin,enrofloxacin,florfenicol and olaquindox,resistant rate were 94.62%,95.38%,92.31%,95.38%,55.38%,57.69%,32.31% and 19.23%,respectively.However,it is more sensitive to tigecycline and vancomycin.The PCR detection showed that 35 strains carried fexA gene,and the detection rate was 26.92%.The 17 strains carried oqxAB genes,and the detection rate was13.08%.In addition,10 strains of oqxAB and fexA genes were also found,and more resistant to most drugs.Among the 130 strains,107 strains can be divided into 21 types of spa,and the spa type t15075(15.38%)is the most common type,followed by t189(13.85%),t034(9.23%),t091(7.69%),t127(6.92%)and t164(6.15%)respectively.In addition,only t034(9.23%)and t3512(3.85%)can be found in strains of three different animals(They are cows,pigs,chickens and cows,pigs,ducks,respectively).In conclusion,oqxABand fexAgenes have been popular in animal farm in Henan province,and carried oqxABand fexAgenes strain resistance is relatively serious,the characteristice of the spatypes was diversity and difference.Therefore,drug resistance and classification monitoring should be strengthened in clinical practice.
The prevalence of oqxAB and fexA genes and the polymorphisms of spa gene were presented to understand the drug resistance of Staphylococcus aureus from different animal sources in Henan province.The sensitivity of the isolated bacteria to 10 antimicrobial agents was determined by the microbroth dilution method,the oqxAB and fexA resistance genes in Staphylococcus aureus were detected by PCR,the X region of spa was studied.The results showed that 130 strains of Staphylococcus aureus were resistant to most drugs,penicillin,erythromycin,clindamycin,tetracycline,ciprofloxacin,enrofloxacin,florfenicol and olaquindox,resistant rate were 94.62%,95.38%,92.31%,95.38%,55.38%,57.69%,32.31% and 19.23%,respectively.However,it is more sensitive to tigecycline and vancomycin.The PCR detection showed that 35 strains carried fexA gene,and the detection rate was 26.92%.The 17 strains carried oqxAB genes,and the detection rate was13.08%.In addition,10 strains of oqxAB and fexA genes were also found,and more resistant to most drugs.Among the 130 strains,107 strains can be divided into 21 types of spa,and the spa type t15075(15.38%)is the most common type,followed by t189(13.85%),t034(9.23%),t091(7.69%),t127(6.92%)and t164(6.15%)respectively.In addition,only t034(9.23%)and t3512(3.85%)can be found in strains of three different animals(They are cows,pigs,chickens and cows,pigs,ducks,respectively).In conclusion,oqxABand fexAgenes have been popular in animal farm in Henan province,and carried oqxABand fexAgenes strain resistance is relatively serious,the characteristice of the spatypes was diversity and difference.Therefore,drug resistance and classification monitoring should be strengthened in clinical practice.
引文
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[16] 张倩,屈勇刚,于会举,等.石河子地区奶牛乳房炎源金黄色葡萄球菌的分离鉴定及耐药性分析[J].黑龙江畜牧兽医,2017(2):102-105.
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[18] 王丽华,吕殿红,张荟,等.质粒介导的喹诺酮耐药基因在动物源金黄色葡萄球菌的流行分布[J].中国兽医学报,2014,34(4):606-612.
[19] 曾淑仪,邓辉,孙坚,等.广东不同地区葡萄球菌cfr基因的流行性调查[J].中国兽医学报,2016,36(8):1376-1382.
[20] FENG Y Y,TIAN W,LIN D C,et al.Prevalence and characterization of methicillin-resistant Staphylococcus pseudintermedius in pets from South China[J].Vet Microbiol,2012,160(3/4):517-524.
[21] 王晓,王新,郝丹,等.奶牛乳源大肠杆菌耐药性、毒素基因鉴定及PFGE分型研究[J].畜牧兽医学报,2015,45(3):463-470.
[2] 林大川,刘健华,王晶,等.猪体内金黄色葡萄球菌及凝固酶阴性葡萄球菌耐药性调查及耐药基因分布研究[J].华南农业大学学报,2012,33(4):550-555.
[3] WU H,LIU B G,LIU J H,et al.Phenotypic and molecular characterization of CTX-M-14 extended-spectrum β-lactamase and plasmid-mediated ACT-like AmpC β-lactamase produced by Klebsiella pneumoniae isolates from chickens in Henan Province,China[J].Genet Mol Res,2012,11(3):3357-3364.
[4] 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.
[5] S?RENSEN A H,HANSEN L H,JOHANNESEN E,et al.Conjugative plasmid conferring resistance to olaquindox[J].Antimicrob Agents Chemother,2003,47(2):798-799.
[6] HANSEN L H,JOHANNESEN E,BURM?LLE M,et al.Plasmid-encoded multidrug efflux pump conferring resistance to olaquindox in Escherichia coli[J].Antimicrob Agents Chemother,2004,48(9):3332-3337.
[7] KEHRENBERG C,SCHWARZ S.fexA,a novel Staphylococcus lentus gene encoding resistance to florfenicol and chloramphenicol[J].Antimicrob Agents Chemother,2004,48(2):615-618.
[8] THE EUROPEAN COMMITTEE ON ANTIMICROBIAL SUSCEPTIBILITY TESTING (EUCAST).Breakpoint tables for interpretation of MICs and zone diameters[EB/OL].2017,Version 7.0.http://www.eucast.org.
[9] HANSEN L H,SORENSEN S J,JORGENSEN H S,et al.The prevalence of the oqxAB multidrug efflux pump amongst olaquindox-resistantEscherichia coli in pigs[J].Microb Drug Resist,2005,11(4):378-382.
[10] CLSI.Performance standards for antimicrobial disk and dilution suseptibility tests for bacteria isolated from animals;approved standard-fouth edition[S].Wayne,PA:Clinical and Laboratory Standards Institute,2013.
[11] PERRETEN V,KADLEC K,SCHWARZ S,et al.Clonal spread of methicillin-resistant Staphylococcus pseudintermedius in Europe and North America:an international multicentre study[J].J Antimicrob Chemother,2010,65(6):1145-1154.
[12] 张静,于三科,王新,等.原料乳和临床乳房炎金黄色葡萄球菌毒力基因检测及药敏分析[J].中国兽医学报,2012,32(5):759-764.
[13] 董翔宸,马友记,张勇,等.奶牛乳腺炎主要病原菌的药物敏感性检测[J].中国兽医学报,2015,35(9):1505-1510.
[14] 杨树鑫,杜军,朱娇玲.奶牛前三把奶中优势病原菌的分离与药敏分析[J].中国兽医杂志,2014,50(11):85-88.
[15] 张强,邢晓楠,王新,等.ST9型动物源耐甲氧西林金黄色葡萄球菌的分子分型、毒素基因检测及耐药性分析[J].畜牧兽医学报,2014,45(5):853-858.
[16] 张倩,屈勇刚,于会举,等.石河子地区奶牛乳房炎源金黄色葡萄球菌的分离鉴定及耐药性分析[J].黑龙江畜牧兽医,2017(2):102-105.
[17] 夏粤华.替加环素对安徽地区4种多重耐药菌体外抗菌活性及其药效学评价[D].安徽合肥:安徽医科大学,2014.
[18] 王丽华,吕殿红,张荟,等.质粒介导的喹诺酮耐药基因在动物源金黄色葡萄球菌的流行分布[J].中国兽医学报,2014,34(4):606-612.
[19] 曾淑仪,邓辉,孙坚,等.广东不同地区葡萄球菌cfr基因的流行性调查[J].中国兽医学报,2016,36(8):1376-1382.
[20] FENG Y Y,TIAN W,LIN D C,et al.Prevalence and characterization of methicillin-resistant Staphylococcus pseudintermedius in pets from South China[J].Vet Microbiol,2012,160(3/4):517-524.
[21] 王晓,王新,郝丹,等.奶牛乳源大肠杆菌耐药性、毒素基因鉴定及PFGE分型研究[J].畜牧兽医学报,2015,45(3):463-470.