临床分离耐甲氧西林溶血葡萄球菌（MRSH）的耐药性及分子流行病学研究
发布时间：2018-08-31 12:31
【摘要】： 第一部分临床分离溶血葡萄球菌中耐甲氧西林菌株(MRSH)及耐药性的检测 前言凝固酶阴性葡萄球菌(coagulase-negative Staphylococcus,CNS)是医院感染的重要病原菌之一,近年来的检出率不断提高。溶血葡萄球菌(Staphylococcus haemolyticus,SH)在凝固酶阴性葡萄球菌中较为常见,检出率一般仅次于表皮葡萄球菌,在某些地区检出率甚至位列第一。耐甲氧西林溶血葡萄球菌(methicillin-resistant Staphylococcus haemolyticus,MRSH)的出现及其快速进化、演变、传播、流行给临床治疗带来极大的困难。MRSH对临床常用的多种抗菌药物具有耐药特征,但对糖肽类抗生素(Glycopeptides)仍然显示敏感,这类抗菌药物的大量应用,已导致了对糖肽类抗菌药物耐药葡萄球菌的出现。为了更好地指导临床用药,我们对安徽省部分医院临床分离的溶血葡萄球菌中MRSH菌株及其耐药性进行了检测和分析。 目的了解安徽省部分医院临床分离的溶血葡萄球菌的耐药性和耐甲氧西林菌株的发生率,为临床合理应用抗菌药物提供依据。 方法收集安徽医科大学第一附属医院、安徽省立医院、皖南医学院附属弋矶山医院、淮北市人民医院及舒城县人民医院等14家医院2005年临床分离的溶血葡萄球菌共103株。按照美国临床实验室标准委员会(Clinical and Laboratory Standards Institute,CLSI)2008年标准,以头孢西丁纸片法及mecA-PCR法检测MRSH,采用琼脂稀释法检测其对常用抗菌药物的耐药性。 结果103株溶血葡萄球菌中,以头孢西丁纸片法及mecA-PCR法,MRSH的检出率分别为86.4%、80.6%,两者无统计学差异(P0.05)。MRSH对万古霉素及替考拉宁极其敏感,未发现耐药菌株;对青霉素、环丙沙星的耐药率80%;对头孢唑啉、头孢呋辛、头孢曲松、克林霉素的耐药率50%;对阿米卡星、四环素、利福平、氯霉素的耐药率分别为16.9%、48.2%、12.0%、31.3%。除阿米卡星、四环素、利福平和氯霉素外,MRSH对其他抗菌药物的耐药率显著高于甲氧西林敏感菌株(methicillin-susceptible Staphylococcus haemolyticus ,MSSH),差异有统计学意义(P0.05)。 结论MRSH对绝大多数抗菌药物呈现高度耐药;对糖肽类抗菌药物仍十分敏感,未发现对糖肽类耐药的溶血葡萄球菌。头孢西丁纸片法和mecA-PCR法检测MRSH无统计学差异(P0.05)。MRSH的检出率高达80%以上,除阿米卡星、四环素、利福平和氯霉素外,MRSH对其他抗菌药物的耐药率均显著高于MSSH(P0.05),临床应加强检测及监测。 第二部分临床分离溶血葡萄球菌中耐甲氧西林菌株(MRSH)的SCCmec分型及同源性分析 前言近年来,临床分离的溶血葡萄球菌中耐甲氧西林菌株(MRSH)的检出率越来越高,许多地区报道已超过80%,耐甲氧西林菌株已经成为了全世界医院感染的重要病原菌。MRSH对临床常用的多种抗菌药物耐药,目前其有效治疗药物主要为糖肽类抗生素:万古霉素、替考拉宁及唑烷酮类抗生素:利奈唑胺。MRSH耐药的主要机制是菌株携带了mecA基因,该基因位于新型的可移动元件SCCmec (Staphylococcal Cassette Chromosomemec mec,SCCmec)基因盒上,该基因盒不仅携带mecA基因,往往还携带有其他抗菌药的耐药基因,从而形成多重耐药。目前已发现5种SCCmec基因型,即SCCmecⅠ、Ⅱ、Ⅲ、Ⅳ和Ⅴ。其中SCCmecⅠ、Ⅱ、Ⅲ型多为医院获得性(HA-MRS),SCCmecⅣ、Ⅴ型多为社区获得性(CA-MRS)。为了了解安徽省临床分离MRSH的SCCmec分型及同源性,我们进行了相关的研究,并报道如下。 目的了解临床分离MRSH的SCCmec分型以及不同型别之间的耐药性差异情况,并对不同SCCmec型别的MRSH进行同源性分析。 方法根据文献,合成引物,多重PCR进行SCCmec分型,ERIC-PCR法对不同SCCmec型别菌株进行同源性分析。 结果1)多重PCR检测SCCmec分型:83株MRSH临床分离菌株中,SCCmecⅠ型23株(27.7 %),SCCmecⅡ型10株(12.1 %),SCCmecⅢ型24株(28.9 %),SCCmecⅣ型1株(1.2 %),Ⅰ、Ⅱ混合型8株(9.6 %),Ⅰ、Ⅲ混合型6株(7.2 %),Ⅱ、Ⅲ混合型5株(6.0 %),Ⅰ、Ⅱ、Ⅲ混合型3株(3.6 %),未分型3株(3.6%);2)不同SCCmec型别菌株的耐药性分析:SCCmecⅠ、Ⅲ型菌株对头孢唑啉、头孢呋辛和头孢曲松的耐药率显著高于Ⅱ型菌株,Ⅱ型菌株对氯霉素的耐药率高于Ⅰ、Ⅲ型菌株(P0.05);3)不同SCCmec型别菌株的同源性分析:23株SCCmecⅠ型分为11型,其中A型5株,B型5株,C型3株,其余8株各为1型,2株未分型;10株SCCmecⅡ型分为6型,其中D型4株,E型2株,3株各为1型,1株未分型;24株SCCmecⅢ型分为9型,其中F型11株,G型2株,H型2株,I型2株,5株各为1型,2株未分型。 结论1)83株MRSH中SCCmecⅠ、Ⅲ型较为多见;2)发现较多的混合型菌株;3)不同型别菌株对部分抗菌药的耐药性有所差异,进一步说明了耐药性与不同SCCmec型别所携带的不同耐药基因有关;4)部分菌株之间存在有克隆传播。
[Abstract]:Part 1 clinical isolates of methicillin-resistant strain (MRSH) and detection of drug resistance in Staphylococcus haemolysin
Coagulase-negative Staphylococcus (CNS) is one of the most important pathogens of nosocomial infection. In recent years, the detection rate of coagulase-negative Staphylococcus haemolyticus (SH) is higher than that of Staphylococcus epidermidis. The emergence of methicillin-resistant Staphylococcus haemolyticus (MRSH) and its rapid evolution, evolution, transmission, epidemic bring great difficulties to clinical treatment. MRSH is resistant to many commonly used antibiotics, but Glycopeptiptide antibiotics. DES) is still susceptible. The wide use of these antibiotics has led to the emergence of glycopeptide-resistant staphylococci. In order to better guide clinical use, MRSH strains and their resistance in clinical isolates from some hospitals in Anhui Province were detected and analyzed.
OBJECTIVE To investigate the drug resistance and the incidence of methicillin-resistant Staphylococcus hemolyticus isolated from some hospitals in Anhui Province, and to provide evidence for rational use of antibiotics.
Methods 103 strains of Staphylococcus hemolyticus were collected from 14 hospitals in 2005, including the First Affiliated Hospital of Anhui Medical University, Anhui Provincial Hospital, Yijishan Hospital Affiliated to Southern Anhui Medical College, Huaibei People's Hospital and Shucheng People's Hospital. E, CLSI) 2008 standard, MRSH was detected by cefoxitin disk method and mecA-PCR method, and the resistance to commonly used antibiotics was detected by agar dilution method.
Results Among 103 strains of hemolytic staphylococcus, the detection rates of MRSH were 86.4% and 80.6% by cefoxitin disk method and mecA-PCR method, respectively. There was no significant difference between the two methods (P 0.05). MRSH was extremely sensitive to vancomycin and teicoplanin, and no drug-resistant strain was found. The resistance rate of MRSH to other antibiotics was significantly higher than that of methicillin-susceptible Staphylococcus haemolyticus (MSSH), except amikacin, tetracycline, rifampicin and chloramphenicol. The difference was statistically significant (P0.05).
Conclusion MRSH is highly resistant to most antibiotics and sensitive to glycopeptide antibiotics, but no hemolytic Staphylococcus was found to be resistant to glycopeptide antibiotics. There is no significant difference between cefoxitin disk method and mecA-PCR method in detecting MRSH (P 0.05). The detection rate of MRSH is over 80%, except amikacin, tetracycline, rifampicin and chloramphenicol. The resistance rate of MRSH to other antibiotics was significantly higher than that of MSSH (P0.05), and clinical detection and monitoring should be strengthened.
Part II SCCMEC typing and homology analysis of methicillin-resistant strains (MRSH) isolated from clinical Staphylococcus hemolyticus
In recent years, the detection rate of methicillin-resistant strains (MRSH) in clinical isolates of Staphylococcus haemolyticus has become higher and higher. More than 80% of MRSH has been reported in many areas. MRSH has become an important pathogen of nosocomial infections all over the world. MRSH is resistant to a variety of antibiotics commonly used in clinic. At present, its effective treatment drugs are mainly sugar. Peptide antibiotics: vancomycin, teicoplanin, and zolidinone antibiotics: linezolid. MRSH resistance is mainly caused by the strain carrying the mecA gene, which is located in the novel mobile element SCCmec (Staphylococcal Cassette Chromosome mec, SCCmec) gene cassette, which carries not only the mecA gene, but also its own. Five SCCmec genotypes, namely SCCmec I, II, III, IV and V, have been identified. SCCmec I, II, III are mostly hospital acquired (HA-MRS) and SCCmec IV, V are mostly community acquired (CA-MRS). Related research is reported and reported below.
Objective To understand the SCCMEC typing of clinical isolated MRSH and the difference of drug resistance among different types of MRSH, and to analyze the homology of different SCCMEC types of MRSH.
Methods According to the literature, primers were synthesized, SCCmec was typed by multiplex PCR, and the homology of different SCCmec strains was analyzed by ERIC-PCR.
Results 1) SCCmec typing was detected by multiplex PCR. Among 83 clinical isolates of MRSH, 23 (27.7%) were SCCmec type I, 10 (12.1%) were SCCmec type II, 24 (28.9%) were SCCmec type III, 1 (1.2%) was SCCmec type IV, 8 (9.6%) were mixed type I and II, 6 (7.2%) were mixed type I and III, 5 (6.0%) were mixed type II, 3 (3.6%) were unclassified. Resistance analysis of different SCCmec strains: SCCmec type I and III strains to cefazolin, cefuroxime and ceftriaxone were significantly higher than type II strains, type II strains to chloramphenicol resistance rate was higher than type I and type III strains (P 0.05); 3) Homology analysis of different SCCmec strains: 23 SCCmec type I strains were divided into 11 types, of which There were 5 strains of type A, 5 strains of type B, 3 strains of type C, 8 strains of type 1 and 2 strains of untyped; 10 strains of SCCmec type I I were divided into 6 types, including 4 strains of type D, 2 strains of type E, 3 strains of type 1 and 1 strain untyped; 24 strains of SCCmec type I I I were divided into 9 types, including 11 strains of type F, 2 strains of type G, 2 strains of type H, 2 strains of type I, 5 strains of type 1 and 2 strains of untyped.
Conclusion 1) SCCmec type I and III were more common in 83 strains of MRSH; 2) More mixed strains were found; 3) Drug resistance of different strains to some antibiotics was different, which further indicated that drug resistance was related to different drug resistance genes carried by different SCCmec types; 4) Clonal transmission was found among some strains.
【学位授予单位】：安徽医科大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R446.5;R181.3
本文编号：2214993
[Abstract]:Part 1 clinical isolates of methicillin-resistant strain (MRSH) and detection of drug resistance in Staphylococcus haemolysin
Coagulase-negative Staphylococcus (CNS) is one of the most important pathogens of nosocomial infection. In recent years, the detection rate of coagulase-negative Staphylococcus haemolyticus (SH) is higher than that of Staphylococcus epidermidis. The emergence of methicillin-resistant Staphylococcus haemolyticus (MRSH) and its rapid evolution, evolution, transmission, epidemic bring great difficulties to clinical treatment. MRSH is resistant to many commonly used antibiotics, but Glycopeptiptide antibiotics. DES) is still susceptible. The wide use of these antibiotics has led to the emergence of glycopeptide-resistant staphylococci. In order to better guide clinical use, MRSH strains and their resistance in clinical isolates from some hospitals in Anhui Province were detected and analyzed.
OBJECTIVE To investigate the drug resistance and the incidence of methicillin-resistant Staphylococcus hemolyticus isolated from some hospitals in Anhui Province, and to provide evidence for rational use of antibiotics.
Methods 103 strains of Staphylococcus hemolyticus were collected from 14 hospitals in 2005, including the First Affiliated Hospital of Anhui Medical University, Anhui Provincial Hospital, Yijishan Hospital Affiliated to Southern Anhui Medical College, Huaibei People's Hospital and Shucheng People's Hospital. E, CLSI) 2008 standard, MRSH was detected by cefoxitin disk method and mecA-PCR method, and the resistance to commonly used antibiotics was detected by agar dilution method.
Results Among 103 strains of hemolytic staphylococcus, the detection rates of MRSH were 86.4% and 80.6% by cefoxitin disk method and mecA-PCR method, respectively. There was no significant difference between the two methods (P 0.05). MRSH was extremely sensitive to vancomycin and teicoplanin, and no drug-resistant strain was found. The resistance rate of MRSH to other antibiotics was significantly higher than that of methicillin-susceptible Staphylococcus haemolyticus (MSSH), except amikacin, tetracycline, rifampicin and chloramphenicol. The difference was statistically significant (P0.05).
Conclusion MRSH is highly resistant to most antibiotics and sensitive to glycopeptide antibiotics, but no hemolytic Staphylococcus was found to be resistant to glycopeptide antibiotics. There is no significant difference between cefoxitin disk method and mecA-PCR method in detecting MRSH (P 0.05). The detection rate of MRSH is over 80%, except amikacin, tetracycline, rifampicin and chloramphenicol. The resistance rate of MRSH to other antibiotics was significantly higher than that of MSSH (P0.05), and clinical detection and monitoring should be strengthened.
Part II SCCMEC typing and homology analysis of methicillin-resistant strains (MRSH) isolated from clinical Staphylococcus hemolyticus
In recent years, the detection rate of methicillin-resistant strains (MRSH) in clinical isolates of Staphylococcus haemolyticus has become higher and higher. More than 80% of MRSH has been reported in many areas. MRSH has become an important pathogen of nosocomial infections all over the world. MRSH is resistant to a variety of antibiotics commonly used in clinic. At present, its effective treatment drugs are mainly sugar. Peptide antibiotics: vancomycin, teicoplanin, and zolidinone antibiotics: linezolid. MRSH resistance is mainly caused by the strain carrying the mecA gene, which is located in the novel mobile element SCCmec (Staphylococcal Cassette Chromosome mec, SCCmec) gene cassette, which carries not only the mecA gene, but also its own. Five SCCmec genotypes, namely SCCmec I, II, III, IV and V, have been identified. SCCmec I, II, III are mostly hospital acquired (HA-MRS) and SCCmec IV, V are mostly community acquired (CA-MRS). Related research is reported and reported below.
Objective To understand the SCCMEC typing of clinical isolated MRSH and the difference of drug resistance among different types of MRSH, and to analyze the homology of different SCCMEC types of MRSH.
Methods According to the literature, primers were synthesized, SCCmec was typed by multiplex PCR, and the homology of different SCCmec strains was analyzed by ERIC-PCR.
Results 1) SCCmec typing was detected by multiplex PCR. Among 83 clinical isolates of MRSH, 23 (27.7%) were SCCmec type I, 10 (12.1%) were SCCmec type II, 24 (28.9%) were SCCmec type III, 1 (1.2%) was SCCmec type IV, 8 (9.6%) were mixed type I and II, 6 (7.2%) were mixed type I and III, 5 (6.0%) were mixed type II, 3 (3.6%) were unclassified. Resistance analysis of different SCCmec strains: SCCmec type I and III strains to cefazolin, cefuroxime and ceftriaxone were significantly higher than type II strains, type II strains to chloramphenicol resistance rate was higher than type I and type III strains (P 0.05); 3) Homology analysis of different SCCmec strains: 23 SCCmec type I strains were divided into 11 types, of which There were 5 strains of type A, 5 strains of type B, 3 strains of type C, 8 strains of type 1 and 2 strains of untyped; 10 strains of SCCmec type I I were divided into 6 types, including 4 strains of type D, 2 strains of type E, 3 strains of type 1 and 1 strain untyped; 24 strains of SCCmec type I I I were divided into 9 types, including 11 strains of type F, 2 strains of type G, 2 strains of type H, 2 strains of type I, 5 strains of type 1 and 2 strains of untyped.
Conclusion 1) SCCmec type I and III were more common in 83 strains of MRSH; 2) More mixed strains were found; 3) Drug resistance of different strains to some antibiotics was different, which further indicated that drug resistance was related to different drug resistance genes carried by different SCCmec types; 4) Clonal transmission was found among some strains.
【学位授予单位】：安徽医科大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R446.5;R181.3
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