臨床分離耐甲氧西林溶血葡萄球菌（MRSH）的耐藥性及分子流行病學研究
發(fā)布時間：2018-08-31 12:31
【摘要】： 第一部分臨床分離溶血葡萄球菌中耐甲氧西林菌株(MRSH)及耐藥性的檢測 前言凝固酶陰性葡萄球菌(coagulase-negative Staphylococcus,CNS)是醫(yī)院感染的重要病原菌之一,近年來的檢出率不斷提高。溶血葡萄球菌(Staphylococcus haemolyticus,SH)在凝固酶陰性葡萄球菌中較為常見,檢出率一般僅次于表皮葡萄球菌,在某些地區(qū)檢出率甚至位列第一。耐甲氧西林溶血葡萄球菌(methicillin-resistant Staphylococcus haemolyticus,MRSH)的出現(xiàn)及其快速進化、演變、傳播、流行給臨床治療帶來極大的困難。MRSH對臨床常用的多種抗菌藥物具有耐藥特征,但對糖肽類抗生素(Glycopeptides)仍然顯示敏感,這類抗菌藥物的大量應用,已導致了對糖肽類抗菌藥物耐藥葡萄球菌的出現(xiàn)。為了更好地指導臨床用藥,我們對安徽省部分醫(yī)院臨床分離的溶血葡萄球菌中MRSH菌株及其耐藥性進行了檢測和分析。 目的了解安徽省部分醫(yī)院臨床分離的溶血葡萄球菌的耐藥性和耐甲氧西林菌株的發(fā)生率,為臨床合理應用抗菌藥物提供依據(jù)。 方法收集安徽醫(yī)科大學第一附屬醫(yī)院、安徽省立醫(yī)院、皖南醫(yī)學院附屬弋磯山醫(yī)院、淮北市人民醫(yī)院及舒城縣人民醫(yī)院等14家醫(yī)院2005年臨床分離的溶血葡萄球菌共103株。按照美國臨床實驗室標準委員會(Clinical and Laboratory Standards Institute,CLSI)2008年標準,以頭孢西丁紙片法及mecA-PCR法檢測MRSH,采用瓊脂稀釋法檢測其對常用抗菌藥物的耐藥性。 結(jié)果103株溶血葡萄球菌中,以頭孢西丁紙片法及mecA-PCR法,MRSH的檢出率分別為86.4%、80.6%,兩者無統(tǒng)計學差異(P0.05)。MRSH對萬古霉素及替考拉寧極其敏感,未發(fā)現(xiàn)耐藥菌株;對青霉素、環(huán)丙沙星的耐藥率80%;對頭孢唑啉、頭孢呋辛、頭孢曲松、克林霉素的耐藥率50%;對阿米卡星、四環(huán)素、利福平、氯霉素的耐藥率分別為16.9%、48.2%、12.0%、31.3%。除阿米卡星、四環(huán)素、利福平和氯霉素外,MRSH對其他抗菌藥物的耐藥率顯著高于甲氧西林敏感菌株(methicillin-susceptible Staphylococcus haemolyticus ,MSSH),差異有統(tǒng)計學意義(P0.05)。 結(jié)論MRSH對絕大多數(shù)抗菌藥物呈現(xiàn)高度耐藥;對糖肽類抗菌藥物仍十分敏感,未發(fā)現(xiàn)對糖肽類耐藥的溶血葡萄球菌。頭孢西丁紙片法和mecA-PCR法檢測MRSH無統(tǒng)計學差異(P0.05)。MRSH的檢出率高達80%以上,除阿米卡星、四環(huán)素、利福平和氯霉素外,MRSH對其他抗菌藥物的耐藥率均顯著高于MSSH(P0.05),臨床應加強檢測及監(jiān)測。 第二部分臨床分離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 前言近年來,臨床分離的溶血葡萄球菌中耐甲氧西林菌株(MRSH)的檢出率越來越高,許多地區(qū)報道已超過80%,耐甲氧西林菌株已經(jīng)成為了全世界醫(yī)院感染的重要病原菌。MRSH對臨床常用的多種抗菌藥物耐藥,目前其有效治療藥物主要為糖肽類抗生素:萬古霉素、替考拉寧及唑烷酮類抗生素:利奈唑胺。MRSH耐藥的主要機制是菌株攜帶了mecA基因,該基因位于新型的可移動元件SCCmec (Staphylococcal Cassette Chromosomemec mec,SCCmec)基因盒上,該基因盒不僅攜帶mecA基因,往往還攜帶有其他抗菌藥的耐藥基因,從而形成多重耐藥。目前已發(fā)現(xiàn)5種SCCmec基因型,即SCCmecⅠ、Ⅱ、Ⅲ、Ⅳ和Ⅴ。其中SCCmecⅠ、Ⅱ、Ⅲ型多為醫(yī)院獲得性(HA-MRS),SCCmecⅣ、Ⅴ型多為社區(qū)獲得性(CA-MRS)。為了了解安徽省臨床分離MRSH的SCCmec分型及同源性,我們進行了相關(guān)的研究,并報道如下。 目的了解臨床分離MRSH的SCCmec分型以及不同型別之間的耐藥性差異情況,并對不同SCCmec型別的MRSH進行同源性分析。 方法根據(jù)文獻,合成引物,多重PCR進行SCCmec分型,ERIC-PCR法對不同SCCmec型別菌株進行同源性分析。 結(jié)果1)多重PCR檢測SCCmec分型:83株MRSH臨床分離菌株中,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型別菌株的耐藥性分析:SCCmecⅠ、Ⅲ型菌株對頭孢唑啉、頭孢呋辛和頭孢曲松的耐藥率顯著高于Ⅱ型菌株,Ⅱ型菌株對氯霉素的耐藥率高于Ⅰ、Ⅲ型菌株(P0.05);3)不同SCCmec型別菌株的同源性分析:23株SCCmecⅠ型分為11型,其中A型5株,B型5株,C型3株,其余8株各為1型,2株未分型;10株SCCmecⅡ型分為6型,其中D型4株,E型2株,3株各為1型,1株未分型;24株SCCmecⅢ型分為9型,其中F型11株,G型2株,H型2株,I型2株,5株各為1型,2株未分型。 結(jié)論1)83株MRSH中SCCmecⅠ、Ⅲ型較為多見;2)發(fā)現(xiàn)較多的混合型菌株;3)不同型別菌株對部分抗菌藥的耐藥性有所差異,進一步說明了耐藥性與不同SCCmec型別所攜帶的不同耐藥基因有關(guān);4)部分菌株之間存在有克隆傳播。
[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.
【學位授予單位】：安徽醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2010
【分類號】：R446.5;R181.3
本文編號：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.
【學位授予單位】：安徽醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2010
【分類號】：R446.5;R181.3
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