CAJ | 학술논문

目的研究上海地区成人患者分离的肺炎链球菌耐药、克隆分型、血清分型、毒力因子携带及生物膜形成等分子特征,为感染控制及治疗提供临床流行病学信息.方法收集2011年1月至2013年12月上海华山医院37株来自门诊和住院成人患者感染分离的非重复肺炎链球菌,用K-B纸片法或E-test法测定对9种常用抗生素(青霉素、万古霉素、红霉素、克林霉素、左氧氟沙星、头孢丙烯、头孢曲松、头孢噻肟、利奈唑胺)的敏感性;用PCR与肺炎链球菌荚膜型血清凝集法确定血清型;采用脉冲场凝胶电泳(PFGE)方法分析不同血清型菌株的基因组特征;采用多位点序列分型(MLST)方法进行克隆分型;采用半定量生物膜形成试验分析其生物膜形成.PCR和凝胶电泳法检测10个主要肺炎链球菌的毒力基因(cbpA、pspA、cps2A、lytA、nanA、pavA、piaA、ply、psaA、spxB).采用Stata统计软件用Fisher's exact test进行相关性统计.结果 37株肺炎链球菌主要血清型包括19F(13.5％),23F(13.5％),14(10.8％),19A(10.8％),青霉素耐药的肺炎链球菌(PRSP)占64.9％.血清型19F、19A、23F与青霉素耐药有明显相关(x2 =5.89,P=0.015)并且都是多重耐药菌(MDR).克隆分型呈多态性,ST81、ST271对包括青霉素在内的抗生素耐药率较高(x2=4.57,P=0.033).生物膜阳性与血清型19A(x2=5.55,P=0.018)、克隆型ST320(x2=4.33,P=0.037)有明显相关,与青霉素耐药等没有明显相关(x2 =0.16,P=0.686).毒力基因检测中lytA、pavA、ply、psaA、spxB均阳性.结论成人肺炎链球菌青霉素耐药率呈上升趋势.特定的血清型、流行克隆分型与抗生素耐药密切相关,为感染控制提供依据.毒力因子PspA等将是未来研制新型疫苗降低肺炎链球菌感染的新型靶标.
목적 연구상해지구성인환자분리적폐염련구균내약、극륭분형、혈청분형、독력인자휴대급생물막형성등분자특정,위감염공제급치료제공림상류행병학신식.방법 수집2011년1월지2013년12월상해화산의원37주래자문진화주원성인환자감염분리적비중복폐염련구균,용K-B지편법혹E-test법측정대9충상용항생소(청매소、만고매소、홍매소、극림매소、좌양불사성、두포병희、두포곡송、두포새우、리내서알)적민감성;용PCR여폐염련구균협막형혈청응집법학정혈청형;채용맥충장응효전영(PFGE)방법분석불동혈청형균주적기인조특정;채용다위점서렬분형(MLST)방법진행극륭분형;채용반정량생물막형성시험분석기생물막형성.PCR화응효전영법검측10개주요폐염련구균적독력기인(cbpA、pspA、cps2A、lytA、nanA、pavA、piaA、ply、psaA、spxB).채용Stata통계연건용Fisher's exact test진행상관성통계.결과 37주폐염련구균주요혈청형포괄19F(13.5％),23F(13.5％),14(10.8％),19A(10.8％),청매소내약적폐염련구균(PRSP)점64.9％.혈청형19F、19A、23F여청매소내약유명현상관(x2 =5.89,P=0.015)병차도시다중내약균(MDR).극륭분형정다태성,ST81、ST271대포괄청매소재내적항생소내약솔교고(x2=4.57,P=0.033).생물막양성여혈청형19A(x2=5.55,P=0.018)、극륭형ST320(x2=4.33,P=0.037)유명현상관,여청매소내약등몰유명현상관(x2 =0.16,P=0.686).독력기인검측중lytA、pavA、ply、psaA、spxB균양성.결론 성인폐염련구균청매소내약솔정상승추세.특정적혈청형、류행극륭분형여항생소내약밀절상관,위감염공제제공의거.독력인자PspA등장시미래연제신형역묘강저폐염련구균감염적신형파표.
Objective Toinvestigate the molecular characteristics including antibiotic resistance,strain type,serotype,virulence,biofilm formation of Streptococcus pneumoniae isolated from Shanghai adult patients.Methods A total of 37 non-repetitive S.pneumoniae isolates causing community acquired and hospital acquired infections of adults were collected from Shanghai Huashan Hospital from January 2011 to December 2013.The inhibitory zone diameter or minimum inhibitory concentrations (MICs) of 9 antimicrobial agents (penicillin,vancomycin,erythromycin,clindamycin,levofloxacin,cefprozi,ceftriaxone,cefotaxime and linezolid) were determined by Kirby-bauer (K-B) method or Etest method;Serotypes were tested by polymerase chain reaction (PCR) and S.pneumoniae antisera agglutination;Genomic characteristics of different serotype strains were determined by pulsed field gel electrophoresis (PFGE)method;Multilocus sequence types (MLST) was used for strain type;Semi quantitative biofilm formation test was used for the biological membrane formation.Ten main pneumococcal virulence genes (cbpA,pspA,cps2A,lytA,nana,pavA,piaA,ply,psaA and spxB) were detected by PCR and gel electrophoresis.Statistical analysis was performed using Stata software and association statistics were tested using Fisher's exact test.Results The most frequent serotypes were 19F (13.5％),23 F (13.5％),14 (10.8％),19A (10.8％).The penicillin resistance rate was 64.9％.Serotypes 19 F,19A and 23 F were significantly associated with penicillin resistance (x2 =5.89,P =0.015) and the isolates belonged to these serotypes were all multi-drug resistant (MDR).ST81 and ST271 showed high resistance rates to several antibiotics including penicillin (x2 =4.57,P =0.033).Biofilm formation was significantly associated with serotypes 19A (x2 =5.55,P =0.018) and strain type ST320 (x2 =4.33,P =0.037),but not associated with penicillin resistance (x2 =0.16,P =0.686).Virulence gene lytA,pavA,ply,psaA,spxB were found in all isolates.Conclusions Penicillin resistance rate of S.pneumoniae in adult is rising.Specific serotype,epidemic clone and antibiotic resistance are closely related,and can provide the basis for the infection control.The virulence factors such as PspA will be the new targets for vaccine development to reduce S.pneumoniae infection in the future.