猪源屎肠球菌tetM基因的检测
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摘要
用微量肉汤倍比稀释法测定了猪源屎肠球菌对四环素等抗生素的敏感性,用PCR和DNA测序方法检测了屎肠球菌中四环素类耐药基因(tet基因),并分析了试验菌对四环素类的耐药机制。结果表明:14株猪源屎肠球菌对四环素、多西环素、环丙沙星、红霉素全部耐药,4株对头孢噻呋耐药;在14株屎肠球菌中,分别有13株和9株含tet L和tet M基因,其中有9株同时携带tet L和tet M基因;猪源屎肠球菌主要存在主动外排和核糖体保护两种耐药机制,其对四环素类的耐药性主要与携带tet M和tet L有关。
The susceptibilities of pig-derived Enterococcus faecium to tetracycline and other antibiotics were detected by using broth microdilution method,and the prevalence of tetracyclines-tolerant genes(tet genes) in Enterococcus faecium was investigated by using PCR and DNA sequencing.The aim of this study is to analyze the tolerant mechanisms of tested Enterococcus faecium strains to tetracyclines.The results showed that all 14 strains of Enterococcus faecium isolated from pigs were tolerant to tetracycline,doxycycline,ciprofloxacin and erythromycin,and 4 strains were tolerant to ceftiofur.Among the above 14 strains,13 strains contained tet L gene,9 strains contained tet M gene,and 9 strains carried both tet L and tet M.The tolerance of pig-derived Enterococcus faecium to tetracyclines was mediated mainly by two mechanisms:drug initiative excretion and ribosomal protection,and it was closely related to carrying tet M and tet L genes.
The susceptibilities of pig-derived Enterococcus faecium to tetracycline and other antibiotics were detected by using broth microdilution method,and the prevalence of tetracyclines-tolerant genes(tet genes) in Enterococcus faecium was investigated by using PCR and DNA sequencing.The aim of this study is to analyze the tolerant mechanisms of tested Enterococcus faecium strains to tetracyclines.The results showed that all 14 strains of Enterococcus faecium isolated from pigs were tolerant to tetracycline,doxycycline,ciprofloxacin and erythromycin,and 4 strains were tolerant to ceftiofur.Among the above 14 strains,13 strains contained tet L gene,9 strains contained tet M gene,and 9 strains carried both tet L and tet M.The tolerance of pig-derived Enterococcus faecium to tetracyclines was mediated mainly by two mechanisms:drug initiative excretion and ribosomal protection,and it was closely related to carrying tet M and tet L genes.
引文
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[4]CLSI.Performance standards for antimicrobial disk and dilution susceptibility testing for bacteria isolated from animals:approved standard[M].Fourth edition.Wayne:Clinical and Laboratory Standards Institute,2013.
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[13]程文海,黄艳,黄澍杰,等.解脲脲原体中tet M基因的检测及其临床应用评价[J].中国中西医结合皮性病学杂志,2005,4(1):37-38.
[14]陆佳,李一经.东北部分地区猪链球菌对四环素类药物耐药机制的研究[J].黑龙江畜牧兽医,2010(1):11-13.
[15]Hu G Z,Pan Y S,Wu H,et al.Prevalence of tetracycline resistance genes and identification of tet(M)in clinical isolates of Escherichia coli from sick ducks in China[J].Journal of Medical Microbiology,2013,62(6):851-858.
[16]Rice L B.Tn916 family conjugative transposons and dissemination of antimicrobial resistance determinants[J].Antimicrob Agents,1998,42:1871-1877.
[17]Courvalin P,Carlier C.Tn1545:a conjugative shuttle transposon[J].Mol Gen Genet,1987,206:259-264.
[18]De Vries L E,Christensen H,Skov R L,et al.Diversity of the tetracycline resistance gene tet(M)and identification of Tn916-and Tn5801-like(Tn6014)transposons in Staphylococcus aureus from humans and animals[J].Journal of Antimicrobial Chemotherapy,2009,64:490-500.
[2]齐景文,予长泳,闫明媚,等.铅黄肠球菌引起雏鸡发病死亡的诊治报告[J].中国家禽,2006,28(19):88.
[3]Agers Y,Pedersen A G,Aarestrup F M.Identification of Tn5397-like and Tn916-like transposons and diversity of the tetracycline resistance gene tet(M)in enterococci from humans,pigs and poultry[J].J Antimicrob Chemother,2006,57:832-839.
[4]CLSI.Performance standards for antimicrobial disk and dilution susceptibility testing for bacteria isolated from animals:approved standard[M].Fourth edition.Wayne:Clinical and Laboratory Standards Institute,2013.
[5]Ng L K,Martin I,Alfa M,et al.Multiplex PCR for the detection of tetracycline resistant genes[J].Molecular and Cellular Probes,2001,15:209-215.
[6]Chopra I,Roberts M.Tetracycline antibiotics:mode of action,applications,molecular biology and epidemiology of bacterial resistance[J].Microbiol Mol Biol Rev,2001,65(2):232
[7]John M C.Roberts update on acquired tetracycline resistance genes[J].FEMS Microbiology Letters,2005,245:195-203.
[8]Thaker M,Spanogiannopoulos P,Wright G.The tetracycline resistance[J].Cell Mol Life Sci,2010,67:419-431.
[9]Warburton P J,Ciric L,Lerner A,et al.Tet AB(46),a predicted heterodimeric ABC transporter conferring tetracycline resistance in Streptococcus australis isolated from the oral cavity[J].J Antimicrob Chemother,2013,68(1):17-22.
[10]Jones C H,Tuckman M,Murphy E,et al.Identification and sequence of a tet(M)tetracycline resistance determinant homologue in clinical isolates of Escherichia coli[J].Journal of Bacteriology,2006,188(20):7151-7164.
[11]Tuckman M,Petersen P J,Howe A Y M,et al.Occurrence of tetracycline resistance genes among E.coli isolates from the phase 3 clinical trials for Tige cycling[J].Antimicrob.Agents Chemother,2007,51(9):3205-3211.
[12]Bryan A,Shapir N,Sadowsky M J.Frequency and distribution of tetracycline resistance genes in genetically diverse,nonselected,and nonclinical Escherichia coli strains isolated from diverse human and animal sources[J].Applied and Environmental Microbiology,2004,70(4):2503-2507.
[13]程文海,黄艳,黄澍杰,等.解脲脲原体中tet M基因的检测及其临床应用评价[J].中国中西医结合皮性病学杂志,2005,4(1):37-38.
[14]陆佳,李一经.东北部分地区猪链球菌对四环素类药物耐药机制的研究[J].黑龙江畜牧兽医,2010(1):11-13.
[15]Hu G Z,Pan Y S,Wu H,et al.Prevalence of tetracycline resistance genes and identification of tet(M)in clinical isolates of Escherichia coli from sick ducks in China[J].Journal of Medical Microbiology,2013,62(6):851-858.
[16]Rice L B.Tn916 family conjugative transposons and dissemination of antimicrobial resistance determinants[J].Antimicrob Agents,1998,42:1871-1877.
[17]Courvalin P,Carlier C.Tn1545:a conjugative shuttle transposon[J].Mol Gen Genet,1987,206:259-264.
[18]De Vries L E,Christensen H,Skov R L,et al.Diversity of the tetracycline resistance gene tet(M)and identification of Tn916-and Tn5801-like(Tn6014)transposons in Staphylococcus aureus from humans and animals[J].Journal of Antimicrobial Chemotherapy,2009,64:490-500.