【摘要】：目的了解肺炎克雷伯菌(Klebsiella pneumoniae,KP)在神經(jīng)重癥加強(qiáng)治療病房(Neurological intensive care unit,NICU)環(huán)境中的存在情況,檢測NICU環(huán)境中及感染患者標(biāo)本中肺炎克雷伯菌質(zhì)粒上攜帶耐藥基因的具體情況,并分析肺炎克雷伯菌質(zhì)粒上攜帶的耐藥基因與菌株耐藥表型的關(guān)系,指導(dǎo)臨床抗菌藥物的使用和NICU環(huán)境的消毒滅菌工作,從而降低NICU中患者肺炎克雷伯菌的感染發(fā)生率。方法對2015年1月-2015年12月在醫(yī)院NICU內(nèi)入住的患者進(jìn)行感染監(jiān)測,并對患者居住的常規(guī)環(huán)境及感染環(huán)境進(jìn)行病原學(xué)監(jiān)測,收集住院感染患者標(biāo)本中及在環(huán)境中定植的肺炎克雷伯菌,檢測其臨床耐藥表型并保留菌株。將收集到的肺炎克雷伯菌接種于LB肉湯培養(yǎng)基中進(jìn)行培養(yǎng),然后使用質(zhì)粒小提試劑盒提取肺炎克雷伯菌內(nèi)的質(zhì)粒DNA,再以質(zhì)粒DNA為模版通過聚合酶鏈反應(yīng)方法對肺炎克雷伯菌菌株質(zhì)粒上攜帶的β-內(nèi)酰胺類、氨基糖苷類和喹諾酮類耐藥基因進(jìn)行檢測。結(jié)果2015年1月-2015年12月期間,在NICU病房環(huán)境中,革蘭氏陽性菌占主導(dǎo)地位,其次為少量的革蘭氏陰性菌和真菌,分布在環(huán)境中的病原菌與NICU內(nèi)感染患者送檢標(biāo)本中檢出的病原菌大致一致。其中,于常規(guī)環(huán)境中檢測到1株肺炎克雷伯菌,并于感染患者居住的感染環(huán)境中檢測出5株肺炎克雷伯菌。研究期間NICU入住患者共259人,其中發(fā)生感染179例,在其送檢的病原學(xué)標(biāo)本中共檢出肺炎克雷伯菌31株。將NICU感染患者送檢標(biāo)本中分離出的31株肺炎克雷伯菌及病房環(huán)境中分離出的6株肺炎克雷伯菌接種于LB肉湯培養(yǎng)基中培養(yǎng)并進(jìn)行細(xì)菌質(zhì)粒DNA的提取,發(fā)現(xiàn)37株肺炎克雷伯菌中均存在質(zhì)粒。同時(shí)對每株肺炎克雷伯菌質(zhì)粒上是否攜帶β-內(nèi)酰胺類、氨基糖苷類和喹諾酮類21種耐藥基因進(jìn)行檢測。感染患者標(biāo)本中分離出的31株肺炎克雷伯菌內(nèi)質(zhì)粒中均檢出SHV和rmt B耐藥基因(100%),而且質(zhì)粒攜帶的耐藥基因均在兩種以上,其中有8株肺炎克雷伯菌的質(zhì)粒同時(shí)攜帶β-內(nèi)酰胺類和氨基糖苷類耐藥基因,其余23株均同時(shí)攜帶β-內(nèi)酰胺類、氨基糖苷類和喹諾酮類耐藥基因。在病房環(huán)境標(biāo)本中分離出的6株肺炎克雷伯菌的質(zhì)粒中也均檢出SHV和rmt B耐藥基因(100%),而且質(zhì)粒攜帶的耐藥基因也均在兩種以上,其中有5株肺炎克雷伯菌的質(zhì)粒同時(shí)攜帶β-內(nèi)酰胺類和氨基糖苷類耐藥基因,僅有1株肺炎克雷伯菌的質(zhì)粒上同時(shí)攜帶β-內(nèi)酰胺類、氨基糖苷類和喹諾酮類耐藥基因。結(jié)論NICU內(nèi)患者感染和環(huán)境中檢出的肺炎克雷伯菌內(nèi)均存在質(zhì)粒,同一質(zhì)粒上可同時(shí)攜帶有SHV、TEM、IMP、rmt B、qnr B、qnr S和acc(6)-Ib-cr等多個(gè)β-內(nèi)酰胺類、氨基糖苷類和喹諾酮類耐藥基因,且每株肺炎克雷伯菌質(zhì)粒上均攜帶有SHV和rmt B類基因,但是肺炎克雷伯菌內(nèi)質(zhì)粒所攜帶的耐藥基因并不與其臨床耐藥表型完全成正比關(guān)系。
[Abstract]:Objective to investigate the presence of Klebsiella pneumoniae (Klebsiella pneumoniae,KP) in (Neurological intensive care unit,NICU, a neurointensive intensive care unit. To detect the drug resistance genes of Klebsiella pneumoniae in NICU environment and infected patients, and to analyze the relationship between drug resistance genes and phenotypes of Klebsiella pneumoniae. In order to reduce the incidence of Klebsiella pneumoniae infection in patients with NICU, we should guide the use of clinical antibiotics and sterilize the environment of NICU. Methods the patients admitted to hospital NICU from January 2015 to December 2015 were monitored for infection, and the routine environment and infection environment were monitored by etiology. Klebsiella pneumoniae (Klebsiella pneumoniae) was collected from inpatients with infection and in the environment to detect the phenotype of clinical drug resistance and to retain the strain. The collected Klebsiella pneumoniae was cultured in LB broth medium, and then the plasmid DNA, was extracted by plasmid extraction kit. The 尾 -lactam, aminoglycoside and quinolone resistance genes carried on the plasmid of Klebsiella pneumoniae were detected by polymerase chain reaction using plasmid DNA as template. Results from January 2015 to December 2015, Gram-positive bacteria dominated the NICU ward environment, followed by a small number of Gram-negative bacteria and fungi. The pathogens distributed in the environment were approximately the same as those detected in the specimens of patients with NICU infection. One strain of Klebsiella pneumoniae was detected in the normal environment and 5 strains of Klebsiella pneumoniae were detected in the infected environment. A total of 259 patients were admitted to NICU during the study, of which 179 cases were infected. A total of 31 strains of Klebsiella pneumoniae were detected in the pathogenic specimens. 31 strains of Klebsiella pneumoniae isolated from NICU infected patients and 6 strains of Klebsiella pneumoniae isolated from ward environment were inoculated in LB broth culture medium to extract bacterial plasmid DNA. Plasmids were found in all 37 strains of Klebsiella pneumoniae. At the same time, 21 resistant genes of 尾-lactams, aminoglycosides and quinolones were detected on the plasmids of Klebsiella pneumoniae. SHV and rmt B resistance genes were detected in 31 strains of Klebsiella pneumoniae isolated from infected patients (100%). Among them, 8 strains of Klebsiella pneumoniae carried 尾 -lactam and aminoglycoside resistance genes simultaneously, and the other 23 strains all carried 尾 -lactam, aminoglycoside and quinolone resistance genes simultaneously. SHV and rmt B resistance genes (100%) were also detected in 6 strains of Klebsiella pneumoniae isolated from ward environmental samples, and the plasmids carried more than two kinds of drug resistance genes. Among them, 5 strains of Klebsiella pneumoniae contained 尾 -lactam and aminoglycoside resistance genes simultaneously, and only one strain of Klebsiella pneumoniae contained 尾 -lactams, aminoglycosides and quinolones on the plasmids. Conclusion there are plasmids in Klebsiella pneumoniae detected in patients with NICU and in the environment. The same plasmid can carry several 尾 -lactams, such as SHV,TEM,IMP,rmt Bnr, acc (6)-Ib-cr and so on. Aminoglycosides and quinolones were resistant genes, and each strain of Klebsiella pneumoniae plasmid carried SHV and rmt B genes, but the drug resistance genes carried in the plasmids of Klebsiella pneumoniae were not directly proportional to the clinical phenotype of drug resistance.
【學(xué)位授予單位】：華北理工大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R446.5

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