铜绿假单胞菌的耐药情况及基因同源性分析

发布时间：2018-03-06 11:31

本文选题：铜绿假单胞菌　切入点：耐药率　出处：《广西医科大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的:1、观察广西医科大学第一附属医院2013年12月~2014年4月检验科细菌实验室从住院患者各类标本中分离培养出的铜绿假单胞菌菌株的来源分布及耐药情况。2、了解该院铜绿假单胞菌菌株之间的亲缘关系。3、对比铜绿假单胞菌的随机扩增多态性DNA(Random amplified polymorphic DNA,RAPD)指纹图谱,判断是否存在局部爆发流行。4、了解铜绿假单胞菌在长期持续感染过程中的RAPD指纹图谱变化情况。方法:1、收集广西医科大学第一附属医院2013年12月~2014年4月检验科细菌实验室从本院住院患者痰液、肺泡灌洗液、血液、尿液、创面分泌物等各类标本中分离培养出的铜绿假单胞菌菌株及其相关信息。用VITEK-2全自动细菌鉴定/药敏分型仪或ATB expression半自动细菌鉴定仪对分离培养出的铜绿假单胞菌菌株进行培养鉴定和药敏分析。细菌培养鉴定和药敏实验遵照《全国临床检验操作规程》的要求进行,同时参照美国临床和实验室标准协会(Clinical and Laboratory Standards Institute,CLSI)2012年制定的标准。分析非重复铜绿假单胞菌的流行病学特征及其药敏情况。2、选择感染铜绿假单胞菌时间大于30天的患者,从他们的痰液、肺泡灌洗液、血液、尿液等标本中分离的铜绿假单胞菌菌株作为本实验的实验菌株。用煮沸法提取细菌基因组DNA,采用RAPD基因分型技术获得RAPD指纹图谱,并进行同源性聚类分析,通过相似性系数了解该院铜绿假单胞菌菌株之间的亲缘关系;对比铜绿假单胞菌的RAPD指纹图谱,判断铜绿假单胞菌局部爆发流行的存在,以及长期持续感染铜绿假单胞菌的过程中,其指纹图谱是否会发生改变。结果:1、共收集铜绿假单胞菌293株。其中剔除同一患者同一部位重复分离菌株后的非重复铜绿假单胞菌为175株。从标本来源来看,非重复铜绿假单胞菌主要分离于患者的呼吸道标本痰及肺泡灌洗液,占52%(91/175),其余依次为创面分泌物占18.9%(33/175)、尿液占6.9%(12/175)、血液占6.3%(11/175)、腹腔引流液占5.7%(10/175),其余标本来自留置导管、咽拭子、组织标本等。从科室分布来看,非重复铜绿假单胞菌主要分离于来自重症监护病房的患者,占25.7%(45/175),其次为烧伤整形外科17.7%(31/175)、中医科占13.1%(23/175)、呼吸内科占8.6%(15/175)和儿科5.1%(9/175),其余标本来自其他临床科室。分析其药敏情况,发现非重复铜绿假单胞菌对氨曲南、环丙沙星、左氧氟沙星、妥布霉素及庆大霉素的耐药率较高,均30%。2、在收集的293株铜绿假单胞菌中,有117株来自感染铜绿假单胞菌持续时间大于30天的患者;这117株铜绿假单胞菌共扩增出19种不同的基因型条带,分别为a~s型,分型率100%。重症监护病房及烧伤整形外科各发生一起在不同患者之间检出相同rapd型别铜绿假单胞菌菌株的情况,其余患者rapd型别各不相同。有4例长期持续感染铜绿假单胞菌的患者,从其痰液、肺泡灌洗液、创面分泌物、血液、尿液等标本中分离培养出的铜绿假单胞菌rapd指纹图谱发生改变,其余19例患者痰液、肺泡灌洗液等送检标本中分离出的铜绿假单胞菌rapd指纹图谱未发生改变。结论:1、铜绿假单胞菌感染主要发生在重症监护病房、烧伤整形外科和中医科;它们对多种抗生素呈不同程度耐药,其中对氨曲南、环丙沙星、左氧氟沙星、妥布霉素及庆大霉素的耐药率均30%,而对多粘菌素b的耐药率为0,敏感率100%。2.铜绿假单胞菌存在基因多态性,部分型别之间具有高度的同源性。3.重症监护病房和烧伤整形外科各发生一起铜绿假单胞菌局部爆发流行。4.铜绿假单胞菌RAPD指纹图谱在其长期持续感染的过程中部分可发生改变。
[Abstract]:Objective: 1 isolates of Pseudomonas aeruginosa strains from various specimens from hospitalized patients in the First Affiliated Hospital of Guangxi Medical University in December 2013 April ~2014 year observation laboratory laboratory source of bacterial distribution and drug resistance of.2, understanding between the hospital Pseudomonas aeruginosa strain relationship.3, random amplified polymorphic DNA comparison of Pseudomonas aeruginosa (the Random amplified polymorphic DNA, RAPD) fingerprint, to determine whether there is local outbreaks of.4, understand the Pseudomonas aeruginosa RAPD fingerprint in long-term changes during infection. Methods: 1, the First Affiliated Hospital of Guangxi Medical University during December 2013 ~2014 April test of bacteria collected from laboratory sputum of patients in this hospital, bronchoalveolar lavage lotion, blood, urine, and related information from Pseudomonas aeruginosa strains from various specimens of wound secretion by VIT. Pseudomonas aeruginosa strain EK-2 automatic bacteria identification / drug sensitive type instrument or ATB expression semi automatic bacteria identification system for isolated culture identification and drug sensitive analysis. Bacteria identification and drug sensitive test in accordance with the "national clinical laboratory operation rules" requirements, with reference to the clinical and laboratory standards (Clinical and Laboratory Standards Institute Association, CLSI) in 2012 to develop a standard. Analysis of non repetitive Pseudomonas aeruginosa epidemiological characteristics and drug sensitivity of.2 infected Pseudomonas aeruginosa for more than 30 days in patients from their sputum, bronchoalveolar lavage fluid, blood and urine of Pseudomonas aeruginosa strains isolated. In the sample as the experimental strains. The bacterial genome DNA was extracted by boiling method, type of technology to obtain RAPD fingerprint by RAPD gene and homology through cluster analysis. A similarity coefficient of understanding the relationship between the strains of Pseudomonas aeruginosa; RAPD fingerprint comparison of Pseudomonas aeruginosa, determine the Pseudomonas aeruginosa outbreak of local existence, and the long-term sustainability of the process of infection of Pseudomonas aeruginosa, the fingerprint is not changed. The results were: 1. The collection of 293 strains of Pseudomonas aeruginosa. The patients with the same site removed the same repeat after non repeated isolates of Pseudomonas aeruginosa and 175 strains. From the specimen source, non repetitive Pseudomonas aeruginosa mainly isolated from respiratory tract specimens of sputum and lavage liquid alveolar patients, accounting for 52% (91/175), followed by the rest wound secretions accounted for 18.9% (33/175), urine accounted for 6.9% (12/175), blood accounted for 6.3% (11/175), liquid drainage accounted for 5.7% (10/175), the specimens from indwelling catheter, throat swab specimens, etc. from the distribution of departments, non repetitive Pseudomonas aeruginosa. Single cell bacteria were mainly isolated from ICU patients, accounted for 25.7% (45/175), followed by burn and plastic surgery 17.7% (31/175), 13.1% (23/175), Department of traditional Chinese medicine, Department of respiratory medicine accounted for 8.6% (15/175) and 5.1% (9/175), the rest of the pediatric specimens from other clinical unit. The analysis of drug sensitivity, non repetitive Pseudomonas aeruginosa to aztreonam, ciprofloxacin, levofloxacin, gentamicin and tobramycin resistance rates were 30%.2, collected in 293 strains of Pseudomonas aeruginosa, 117 strains of Pseudomonas aeruginosa from infection lasted for more than 30 days of patients; the 117 strains of Pseudomonas aeruginosa were amplified 19 different genotype bands were a~s type, 100%. type rate of plastic surgery in ICU and burn all occurred in the same type of RAPD detection of Pseudomonas aeruginosa strains in different patients among the remaining patients, the type of RAPD The same. 4 cases of persistent infection of Pseudomonas aeruginosa in patients from the sputum, bronchoalveolar lavage fluid, wound secretions, blood, cultivate the Pseudomonas aeruginosa RAPD fingerprint changed separation of urine specimens, the remaining 19 cases of patients with sputum, bronchoalveolar lavage fluid and isolated specimens sent in P. Pseudomonas RAPD fingerprint is not changed. Conclusion: 1, mainly occurred in the ICU of Pseudomonas aeruginosa infection, burns and plastic surgery and Department of traditional Chinese medicine; they showed different degrees of resistance to multiple antibiotics, including of aztreonam, ciprofloxacin, levofloxacin, gentamicin and tobramycin resistant rate was 30%, and the the polymyxin B resistance rate was 0, the sensitive rates of Pseudomonas aeruginosa 100%.2. gene polymorphism, partial type is highly homologous to.3. in ICU and burn plastic surgery occurred in the P. The local outbreak of Pseudomonas aeruginosa (.4.) RAPD fingerprint of Pseudomonas aeruginosa can be partially altered in the process of long-term persistent infection.

【学位授予单位】：广西医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R446.5

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