2018—2022年某院高毒力碳青霉烯类耐药肺炎克雷伯菌流行病学特点及耐药性分析

Epidemiological characteristics and drug resistance of hypervirulent carbapenem-resistant Klebsiella pneumoniae in a hospital,2018-2022

目的分析2018—2022年某院高毒力碳青霉烯类耐药肺炎克雷伯菌(carbapenem-resisitant hypervirulent Klebsiella pneumoniae,CR-hvKP)流行病学特点及耐药性。方法收集2018年1月—2022年12月山东省立第三医院分离的100株碳青霉烯类耐药肺炎克雷伯菌(carbapenem-resisitant Klebsiella pneumoniae,CR-KP),筛选出CR-hvKP。采用K-B纸片法进行药敏试验;聚合酶链式反应(polymerase chain reaction,PCR)检测CR-hvKP的多位点序列分型(multilocus sequence typing,MLST)、CR-hvKP分离株毒力基因;肠杆菌基因间重复一致序列-聚合酶链反应(enterobacterial repetitive intergenic consensus sequence-polymerase chain reaction,ERIC-PCR)DNA指纹图谱检测CR-hvKP分离菌株的同源性。结果从100株CR-KP中分离出15株CR-hvKP(15.00%),患者年龄45~89岁;男7例,女8例;来自重症科室7株(46.67%);15株CR-hvKP分离株对黏菌素、替加环素、甲氧苄啶-磺胺甲恶唑较为敏感外,对其余抗生素的耐药率均>70%。MLST分型结果显示,13株CR-hvKP分离株为ST11,ST23、ST86各1株;荚膜血清分型结果显示,8株CR-hvKP分离株为K1型,5株为K2型,K64、K47各1株;耐药基因检测结果显示,13株携带KPC-2(86.67%),5株携带NDM-1(33.33%),1株携带IMP-1(6.67%),其中有4株同时携带KPC-2、NDM-1基因。15株CR-hvKP分离株均携带entB、fimH、rmpA2、rmpA、iucA基因(100.00%),14株携带iroB基因,13株携带iroN、mrkD基因,3株携带aerobactin基因。3株携带毒力基因9种,9株携带毒力基因8种,3株携带毒力基因7种。经PCR扩增和琼脂糖凝胶电泳后,出现3~9个DNA片段。DNA指纹图谱结果显示,15株CR-hvKP分离株具有高度多态性。结论2018—2022年某院CR-hvKP的检出率为15.00%,来源多为重症科室,对多种抗生素耐药,对黏菌素、替加环素较为敏感,且携带多种耐药基因及毒力基因,具有高度多态性。

Objective To analyze the epidemiological features and drug resistance of carbapenem-resistant hypervirulent Klebsiella pneumoniae(CR-hvKP)in a hospital from 2018 to 2022.Methods A total of 100 strains of carbapenem-resistant Klebsiella pneumoniae(CR-KP)isolated from the Third Hospital of Shandong Province from January 2018 to December 2022 were collected and screened for CR-hvKP.The K-B paper method was used for drug sensitivity test.Polymerase chain reaction(PCR)was employed to detect the multilocus sequence typing(MLST)and virulence gene of CR-hvKP isolates.Enterobacterial repetitive intergenic consensus sequence-polymerase chain reaction(ERIC-PCR)DNA fingerprinting was applied to detecting the homology of CR-hvKP isolates.Results Fifteen(15.00%)strains of CR-hvKP were isolated from 100 strains of CR-KP.The patients were aged 45-89 years,including 7 males and 8 females.7(46.67%)strains were isolated from the intensive care unit.Except for being more sensitive to colistin,tigecycline and trimethoprim-sulfamethoxazole,the resistance rates of the 15 CR-hvKP isolates to other antibiotics listed in Table 2 were all>70%.MLST typing results displayed that 13 CR-hvKP isolates were ST11,1 CR-hvKP isolate was ST23 and 1 CR-hvKP isolate ST86.Capsular serotyping results revealed that 8 CR-hvKP isolates were K1 type,5 CR-hvKP isolates K2 type,1 CR-hvKP isolate was K64,and 1 CR-hvKP isolate K47.Drug resistance gene detection results showed that 13 CR-hvKP isolates carried KPC-2(86.67%),5 CR-hvKP isolates NDM-1(33.33%),and 1 CR-hvKP isolate IMP-1(6.67%),among which 4 carried both KPC-2 and NDM-1 genes.All 15 CR-hvKP isolates carried entB,fimH,rmpA2,rmpAand iucAgenes(100.00%),14 CR-hvKP isolates iroB gene,13 CR-hvKP isolates iroN and mrkD genes,and 3 CR-hvKP isolates aerobactin gene.3 isolates carried 9 virulence genes,9 isolates 8 virulence genes,and 3 isolates 7 virulence genes.After PCR amplification and agarose gel electrophoresis,3-9 DNA fragments appeared.DNA fingerprinting results demonstrated that the 15 CR-hvKP isolates had high polymorphism.Conclusion The detection rate of CR-hvKP in the hospital from 2018 to 2022 was 15.00%,mostly from the intensive care unit.The 15 CR-hvKP isolates detected are resistant to multiple antibiotics and more sensitive to colistin and tigecycline,carry multiple drug resistance genes and virulence genes,and have high polymorphism.