淮安市56株临床分离肺炎克雷伯菌耐药性和分子分型特征
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摘要
目的了解淮安市临床分离肺炎克雷伯菌的耐药特征以及分子分型特点。方法 2018年1—6月,收集国家致病菌识别网络淮安市各哨点医院从临床标本中分离的肺炎克雷伯菌株,以拉丝试验测定分离菌株的黏液表型,以微量肉汤稀释法测定菌株对24种常用抗生素的耐药性,PCR检测荚膜血清型、毒力基因和碳青霉烯类耐药基因,利用脉冲场凝胶电泳进行分子分型。结果共收集临床分离肺炎克雷伯菌56株,高毒力肺炎克雷伯菌(hvKP)占55.36%,经典肺炎克雷伯菌(cKP)占44.64%。菌株多来自脓肿液(26.79%)、痰液(41.07%)。总耐药率为66.07%,多重耐药菌占58.93%,ESBLs菌株占17.86%,耐碳青霉烯类抗生素(美罗培南、亚胺培南)菌株占35.71%,3株为多粘菌素耐药菌株。除氯霉素、吉米沙星外,高毒力肺炎克雷伯菌(hvKP)MIC50均>经典肺炎克雷伯菌(cKP)。耐药基因kpc的检出率为28.57%,cKP菌株的携带率(48.00%)大于hvKP菌株(12.90%),差异有统计学意义(P=0.004),OXA-48和NDM-1未检出。K1、K2、K5、K20、K54、K57荚膜血清型检出率分别为16.07%、10.71%、3.57%、8.93%、9.68%和3.23%。毒力基因magA、rmpA在hvKP(29.03%,80.65%)中检出率均高于cKP菌株(4.00%,24.00%),差异均有统计学意义(P值均<0.05)。菌株PFGE带型相似度为5.91%～100.00%。结论淮安市临床分离肺炎克雷伯菌多为高毒力菌株,且耐药水平较高,PFGE带型多样性较高,应加强监测及临床抗生素使用指导。
Objective To investigate the drug resistance and molecular characteristics of clinical isolates of Klebsiella pneumoniae in Huai'an.Methods Klebsiella pneumoniae strains were collected from pathogen identification network from Jan to Jun in 2018.The phenotype of produce mucus was determined by string test.Drug resistance to 24 antibiotics was detected by MIC method.Capsular serotypes,virulent genes and carbapenem resistant genes were detected by PCR.Molecular subtyping traits of isolated strains were characterized by PFGE.Results A total of the 56 Klebsiella pneumonia strains isolated from clinics were collected,the prevalence of high virulence Klebsiella pneumonia(hvKP)was 55.36%,the classic klebsiella pneumoniae(cKP)accounted for 44.64%.The isolated strains were mainly from abscess(26.79%)and sputum(41.70%)samples.The overall drug resistance of isolated strains were 66.07%,the multi-drug resistant strains accounted for 58.93%,ESBLs isolates accounted for 17.86%,carbapenems(imipenem and meropenem)resistant strains accounted for 35.71%;3 polymyxin resistant strains were identified.Except of chloramphenicol and jimmy floxacin chloramphenicol,the MIC50 values to tested antibiotics of hvKP were higher than those of cKP.The detection rate of kpc gene was 28.57%;the prevalence of kpc gene in cKP strains(48.00%)was higher than that of hvKP strains(12.90%),the difference was statistically significant(P=0.004).The OXA-48 and NDM-1 genes were not detected.The incidences of capsular serotye K1,K2,K5,K20,K54 and K57 were 16.07%,10.71%,3.57%,8.93%,9.68% and 3.23%,respectively.The detection rates of magA and rmpA of hvKP(29.03%,80.65%)were both higher than those of cKP(4.00%,24.00%),the differences were statistically significant(all P<0.05).The similarity of PFGE bands were 5.91%-100.00%.Conclusion The Klebsiella pneumoniae isolates from clinical in this region were mostly high virulent strains with high drug resistance and high diversity of PFGE band patterns.It was necessary to strengthen monitoring,clinical guidance and control of antibiotic usage.
Objective To investigate the drug resistance and molecular characteristics of clinical isolates of Klebsiella pneumoniae in Huai'an.Methods Klebsiella pneumoniae strains were collected from pathogen identification network from Jan to Jun in 2018.The phenotype of produce mucus was determined by string test.Drug resistance to 24 antibiotics was detected by MIC method.Capsular serotypes,virulent genes and carbapenem resistant genes were detected by PCR.Molecular subtyping traits of isolated strains were characterized by PFGE.Results A total of the 56 Klebsiella pneumonia strains isolated from clinics were collected,the prevalence of high virulence Klebsiella pneumonia(hvKP)was 55.36%,the classic klebsiella pneumoniae(cKP)accounted for 44.64%.The isolated strains were mainly from abscess(26.79%)and sputum(41.70%)samples.The overall drug resistance of isolated strains were 66.07%,the multi-drug resistant strains accounted for 58.93%,ESBLs isolates accounted for 17.86%,carbapenems(imipenem and meropenem)resistant strains accounted for 35.71%;3 polymyxin resistant strains were identified.Except of chloramphenicol and jimmy floxacin chloramphenicol,the MIC50 values to tested antibiotics of hvKP were higher than those of cKP.The detection rate of kpc gene was 28.57%;the prevalence of kpc gene in cKP strains(48.00%)was higher than that of hvKP strains(12.90%),the difference was statistically significant(P=0.004).The OXA-48 and NDM-1 genes were not detected.The incidences of capsular serotye K1,K2,K5,K20,K54 and K57 were 16.07%,10.71%,3.57%,8.93%,9.68% and 3.23%,respectively.The detection rates of magA and rmpA of hvKP(29.03%,80.65%)were both higher than those of cKP(4.00%,24.00%),the differences were statistically significant(all P<0.05).The similarity of PFGE bands were 5.91%-100.00%.Conclusion The Klebsiella pneumoniae isolates from clinical in this region were mostly high virulent strains with high drug resistance and high diversity of PFGE band patterns.It was necessary to strengthen monitoring,clinical guidance and control of antibiotic usage.
引文
[1]杨帆,邓保国,魏纪东,等.医源与动物源肺炎克雷伯菌耐药性和分子特性研究[J].中国人兽共患病学报,2017,33(10):888-902.
[2]林星宇,王印,杨泽晓,等.猪源肺炎克雷伯菌的分离鉴定[J].中国预防兽医学报,2015,37(5):375-378.
[3] Decre D,Verdet C,Emirian A,et al.Emerging severe and fatal infections due to Klebsiella pneumoniae in two university hospitals in France[J].J Clin Microbiol,2011,49(8):3012-3014.
[4]田李均,王晓丽,肖淑珍,等.医院内高黏液性肺炎克雷伯菌的流行分布、毒力基因及临床特征分析[J].上海交通大学学报(医学版),2017,37(1):43-48.
[5] Pomakova DK,Hsiao CB,Beanan JM,et al.Clinical and phenotypic differences between classic and hypervirulent Klebsiella pneumonia:an emerging and under-recognized pathogenic variant[J].Eur J Clin Microbiol Infect Dis,2012,31(6):981-989.
[6]方柳,杨珍,石源平,等.浅述动物源性肺炎克雷伯菌病流行情况及相关致病因子[J].江西畜牧兽医杂志,2017,36(6):4-7.
[7]白耀霞,杨思迪,徐杰.肺炎克雷伯菌耐药性和毒力相关因子研究进展[J].江西医药,2017,52(5):469-470.
[8]顾丹霞.不同人群肠道肺炎克雷伯菌毒力及耐药情况研究[D].杭州:浙江大学,2015.
[9] Cockerill FR,Patel JB,Alder J,et al.Performance standards for antimicrobial susceptibility testing:twenty-third informational sup-plement M100-S24[Z].
[10]葛婧,钟雪珊,肖刚,等.广州地区急性腹泻患儿粪便中多重耐药及高毒力肺炎克雷伯菌分布检测[J].中华疾病控制杂志,2017,21(6):562-566.
[11]沈定霞,李东冬,郭玲,等.高黏液表型肺炎克雷伯菌的荚膜分型与毒力基因的检测[J].中华检验医学杂志,2014,37(5):379-382.
[12] Zhang R,Lin D,Chan EW,et al.Emergence of carbapenemresistant serotype K1 hypervirulent Klebsiella pneumoniae(hvKP)strains in China[J].Antimicrob Agents Chemother,2015,60(1):709-711.
[13] Yao B,Xiao X,Wang F,et al.Clinical and molecular characteristics of multi-clone carbapenem-resistant hypervirulent(hypermucoviscous)Klebsiella pneumoniae isolates in a tertiary hospital in Beijing,China[J].Int J Infect Dis,2015,37:107-112.
[14] Li W, Sun G, Yu Y,et al.Increasing occurrence of antimicrobial-resistant hypervirulent(hypermucoviscous)Klebsiella pneumoniae isolates in China[J].Clin Infect Dis,2014,58(2):225-232.
[15] Shon AS,Bajwa RP,Russo TA.Hypervirulent(hypermucoviscous)Klebsiella pneumoniae:a new and dangerous breed[J].Virulence,2013,4(2):107-118.
[16] Zhang Y,Zhao C,Wang Q,et al.High prevalence of hypervirulent Klebsiella pneumoniae infection in China:geographic distribution,clinical characteristicsand antimicrobial resistance[J].Antimicrob Agents Chemother,2016,60(10):6115-6120.
[17]吕国平,李丽婕,芦飞,等.食源性金黄色葡萄球菌脉冲场凝胶电泳分型分析[J].微生物学杂志,2013,33(4):55-58.
[2]林星宇,王印,杨泽晓,等.猪源肺炎克雷伯菌的分离鉴定[J].中国预防兽医学报,2015,37(5):375-378.
[3] Decre D,Verdet C,Emirian A,et al.Emerging severe and fatal infections due to Klebsiella pneumoniae in two university hospitals in France[J].J Clin Microbiol,2011,49(8):3012-3014.
[4]田李均,王晓丽,肖淑珍,等.医院内高黏液性肺炎克雷伯菌的流行分布、毒力基因及临床特征分析[J].上海交通大学学报(医学版),2017,37(1):43-48.
[5] Pomakova DK,Hsiao CB,Beanan JM,et al.Clinical and phenotypic differences between classic and hypervirulent Klebsiella pneumonia:an emerging and under-recognized pathogenic variant[J].Eur J Clin Microbiol Infect Dis,2012,31(6):981-989.
[6]方柳,杨珍,石源平,等.浅述动物源性肺炎克雷伯菌病流行情况及相关致病因子[J].江西畜牧兽医杂志,2017,36(6):4-7.
[7]白耀霞,杨思迪,徐杰.肺炎克雷伯菌耐药性和毒力相关因子研究进展[J].江西医药,2017,52(5):469-470.
[8]顾丹霞.不同人群肠道肺炎克雷伯菌毒力及耐药情况研究[D].杭州:浙江大学,2015.
[9] Cockerill FR,Patel JB,Alder J,et al.Performance standards for antimicrobial susceptibility testing:twenty-third informational sup-plement M100-S24[Z].
[10]葛婧,钟雪珊,肖刚,等.广州地区急性腹泻患儿粪便中多重耐药及高毒力肺炎克雷伯菌分布检测[J].中华疾病控制杂志,2017,21(6):562-566.
[11]沈定霞,李东冬,郭玲,等.高黏液表型肺炎克雷伯菌的荚膜分型与毒力基因的检测[J].中华检验医学杂志,2014,37(5):379-382.
[12] Zhang R,Lin D,Chan EW,et al.Emergence of carbapenemresistant serotype K1 hypervirulent Klebsiella pneumoniae(hvKP)strains in China[J].Antimicrob Agents Chemother,2015,60(1):709-711.
[13] Yao B,Xiao X,Wang F,et al.Clinical and molecular characteristics of multi-clone carbapenem-resistant hypervirulent(hypermucoviscous)Klebsiella pneumoniae isolates in a tertiary hospital in Beijing,China[J].Int J Infect Dis,2015,37:107-112.
[14] Li W, Sun G, Yu Y,et al.Increasing occurrence of antimicrobial-resistant hypervirulent(hypermucoviscous)Klebsiella pneumoniae isolates in China[J].Clin Infect Dis,2014,58(2):225-232.
[15] Shon AS,Bajwa RP,Russo TA.Hypervirulent(hypermucoviscous)Klebsiella pneumoniae:a new and dangerous breed[J].Virulence,2013,4(2):107-118.
[16] Zhang Y,Zhao C,Wang Q,et al.High prevalence of hypervirulent Klebsiella pneumoniae infection in China:geographic distribution,clinical characteristicsand antimicrobial resistance[J].Antimicrob Agents Chemother,2016,60(10):6115-6120.
[17]吕国平,李丽婕,芦飞,等.食源性金黄色葡萄球菌脉冲场凝胶电泳分型分析[J].微生物学杂志,2013,33(4):55-58.