Molecular epidemiology of penicillin-resistant Streptococcus pneumoniae in a university hospital, Ankara, Turkey

Penicillin-resistant Streptococcus pneumoniae isolates (n = 76) from clinical samples of patients admitted to Hacettepe University Hospital between January 1997 and December 2001 were included in the study. MICs of penicillin G, erythromycin A, clindamycin, cefaclor, cefotaxime, vancomycin, chloramphenicol, tetracycline, ciprofloxacin and rifampicin were determined by agar dilution. The isolates were serogrouped on the basis of the Neufeld Quellung reaction and were typed by BOX-PCR. Genetic polymorphism of the penicillin resistance genes pbp2b and pbp2x was investigated by restriction fragment length polymorphism (RFLP) analysis. Of the 76 isolates tested, 64 (84.2%) showed intermediate resistance to penicillin, while 12 (15.8%) were resistant to higher levels of penicillin (MIC > or = 2 mg/L). The resistance patterns of the isolates revealed six different resistance profiles. There were 22 different serotypes, with c. 55% of the isolates belonging to serotypes 23B, 19A, 19F, 14, 6 A and 9V. Five distinct patterns for pbp2b and 12 distinct patterns for pbp2x were obtained by RFLP analysis of penicillin-binding protein genes. The combination of these patterns allowed isolates to be classified into 22 fingerprint subgroups. BOX-PCR analysis showed that the isolates fell into 14 distinct BOX genotypes, with 33 subtypes. Serotype 9V isolates with pbp genotype 2-6 and BOX-PCR type 4, 4.1 or 4.2 were related to the pandemic clone Spain(9V)-3. No relatedness to other international clones was detected among the other study strains, but genetic relatedness was observed among some of the serotype 19A and 23B isolates. Overall, the results demonstrated that most of the penicillin-resistant pneumococcal isolates in Turkey, other than those belonging to serotypes 9V, 19A and 23B, were derived from several independent clones, possibly resulting from multiple importation of strains originating from outside the country. Differences in pbp patterns, serotypes and resistance profiles among isolates that showed similar BOX-PCR patterns supported the hypothesis that horizontal transfer of capsular genes, pbp genes and other genetic determinants between S. pneumoniae and viridans group streptococci may have occurred.     

In vivo efficacy of cefotaxime and amoxicillin against penicillin-susceptible, penicillin-resistant and penicillin-cephalosporin-resistant strains of Streptococcus pneumoniae in a mouse pneumonia model

Objective: To compare cefotaxime (CTX) to amoxicillin (AMO) (usually considered the definitive therapy for penicillinsusceptible Streptococcus pneumoniae infections) in an immunocompromised mouse pneumonia model.
Methods: Three S. pneumoniae clinical isolates were used: two serotype 19 strains, a penicillin-susceptible (P^s) strain (penicillin MIC = 0.03 μ/mL) and a highly penicillin-resistant (P^r) strain (penicillin MIC = 4 μ/mL), and one serotype 23F strain, a penicillin-cephalosporin-resistant (CFTR) strain (CTX MIC = 4 μ/mL).
Results: CTX activity in this mouse model of pneumonia induced by the highly penicillin-resistant strain of S. pneumoniae was lower than expected from its low MIC against this organism. Furthermore, AMO had greater efficacy than CTX against a CFTR S. pneumoniae strain.
Conclusion: Our data suggest that there is no major difference in the in vivo efficacy of the two agents, cefotaxime and amoxicillin, against penicillin-resistant and penicillin-cephalosporin-resistant S. pneumoniae.     

成都地区肺炎链球菌对抗菌药物的耐药性调查

目的 了解成都地区临床分离的肺炎链球菌的耐药性，为肺炎链球菌感染临床合理应用抗菌药物提供理论依据。方法 二倍琼脂稀释法测定11种抗菌药物对肺炎链球菌的最低抑菌浓度(MIC)。结果 91．94％菌株对青霉素敏感，8．06％中度耐药；88．71％菌株对SMZ／TMP敏感，11．29％中度耐药；肺炎链球菌对头孢呋辛、头孢噻肟、头孢毗肟、氧氟沙星、司帕沙星、美洛培南、万古霉素敏感率为100％；对红霉素、克林霉素耐药率相当高，分别达到62．90％和74．19％。结论 成都地区肺炎链球菌对青霉素耐药率较低，而对大环内酯类和克林霉素类耐药率较高。     

Clinical outcome of invasive infections in children caused by highly penicillin-resistant Streptococcus pneumoniae compared with infections caused by penicillin-susceptible strains

BACKGROUND: In this report based on data from the Institutional Surveillance System during 1994-1998, we document the continuing emergence of drug-resistant Streptococcus pneumoniae strains at the Hospital Infantil de Mexico Federico Gómez in Mexico City. METHODS: We evaluate the clinical course of 49 invasive pneumococcal infection outside the central nervous system (CNS) by a number of factors including the site, severity, and place where the infection was acquired, the underlying health of the patient, and the adequacy of antimicrobial therapy. RESULTS: An underlying illness was present in 21 of 49 (43%) patients, 37 (75%) patients had taken previous antimicrobial therapy, and 25% of the infections were nosocomially acquired. Overall, 25 of 49 (51%) of the pneumococcal strains tested were pencillin-resistant; strains with the highest resistance to penicillin were also resistant to cephalosporins. Twenty-two percent of all strains were considered to be multidrug-resistant. Eleven of 25 penicillin-resistant strains were identified as multidrug-resistant, i.e., to erythromycin, TMP/SMX, and chloramphenicol. Ten serotypes accounted for 88% of the isolates, the most frequent serotypes being 23F, 14, 19V, 6A, and 6B. The overall case-fatality rate was 37% (18 of 49), with most deaths occurring within 3-5 days after antibiotic therapy was initiated. There was no difference in the case fatality rate between children with penicillin-nonsusceptible and penicillin-susceptible pneumococcal infections; instead; case-fatality rate correlated with severity of illness on admission and presence of underlying disease. CONCLUSIONS: Characterizing groups at risk for invasive pneumococcal disease could aid in the development of preventive programs and increase the benefits from wide use of future conjugated vaccines.     

肺炎链球菌对大环内酯类抗生素耐药机制研究

目的观察耐大环内酯类肺炎链球菌的耐药表型和基因型。方法用琼脂二倍稀释法测定耐大环内酯类肺炎链球菌对11种抗生素的最低抑菌浓度,耐药表型由红霉素和克林霉素双纸片法初步分型,通过PCR扩增耐药基因ermB和mefA进行基因分型,并测定ermB和mefA基因序列。结果　所有23株肺炎链球菌对大环内酯和克林霉素高度耐药,均表现为内在型耐药,没有发现诱导型耐药和M型耐药。ermB基因在23株肺炎链球菌中均检测到,其中5株同时检测到mefA基因,没有发现仅单一mefA基因阳性的菌株,基因型与耐药表型完全一致。所测ermB和mefA基因序列与基因库收录序列高度一致。 结论ermB基因介导的靶位改变可能是重庆地区肺炎链球菌对大环内酯类抗生素的主要耐药机制。     

变异链球菌的微生态学研究进展

随着微生态学的不断发展,医学微生态学的研究越来越受到重视。变异链球菌是人类的主要致龋菌,通过黏附、产酸和耐酸导致牙体组织破坏,从而形成龋齿。本文就变异链球菌的感染传播途径、生物信号传导系统、遗传多态性、致龋性及免疫防龋的研究进展作一综述。     

中药五倍子对口腔致龋菌影响的体外实验研究

目的：研究五倍子对致龋菌生长、产酸及产胞外多糖的影响，探讨五倍子是否能有效调节口腔菌群生态平衡。方法：测定五倍子对3种主要致龋菌－变形链球菌、粘性放线菌和血链球菌的最低抑菌浓度MIC。再测定低于MIC的4个浓度的五倍子对3种细菌产酸及产生水不溶性多糖能力的影响。结果：五倍子对3种细菌的生长、产酸均有一定的抑制作用，且能够有效抑制变形链球菌产生水不溶性葡聚糖。结论：五倍子能有效抑制变形链球菌、粘性放线菌和血链球菌的生长、产酸及抑制变形链球菌产水不溶性葡聚糖。     

2004～2009年临床分离肺炎链球菌的血清型/群分布及耐药性监测

目的：了解当前本院临床分离肺炎链球菌的血清型/群分布及耐药趋势,为临床合理使用抗生素提供参考。方法：以2004～2009年临床分离的822株肺炎链球菌为研究对象,采用荚膜肿胀试验进行血清分型/群,E-test检测菌种对青霉素、头孢呋辛、头孢地尼、头孢克罗、红霉素、四环素、左氧氟沙星、万古霉素等8种抗生素的敏感性。结果：2004年青霉素不敏感肺炎链球菌（PNSP）的分离率为49.4%,并呈逐年上升趋势,至2006年PNSP分离率高达67.8%,2009年下降至51.0%。822株肺炎链球菌最常见的型/群是19群,其次是23、6、14、3、其他。PNSP在6种血清型/群中所占的比例在2004～2009年期间无显著性变化（P〉0.05）。肺炎链球菌对其他β内酰胺类抗生素的非敏感趋势类似青霉素,对红霉素、四环素的非敏感率始终在60%以上,其中青霉素耐药肺炎链球菌（PRSP）对红霉素和四环素几乎100%耐药,对左氧氟沙星、万古霉素的非敏感率均〈3%。结论：临床分离肺炎链球菌以19群、23群、6群、14群、3群常见;对β内酰胺类抗生素的非敏感率自2007年呈下降趋势,且对左氧氟沙星、万古霉素始终具有较高的敏感性。     

肺炎链球菌肺炎家兔模型的建立及实验研究

 目的实验旨在建立肺炎链球菌感染的家兔肺炎模型,对肺炎链球菌的发病机制进行初步的探讨,为药物治疗提供理论基础,为药学及药物治疗的研究建立肺炎链球菌家兔模型。方法雄性新西兰兔20只,正常对照组7只,实验组13只。在全麻下经气管插管给实验组家兔注射1 mL浓度为10¹⁰cfu·mL^-1的肺炎链球菌。接菌后,分别于0,4,6,30,54,76 h测体温和白细胞,并于76 h后处死全部动物,计算实验过程中的死亡率。取动物肺脏进行大体标本观察,制作肺组织切片并镜下观察。将肺组织匀浆做细菌培养。正常对照组也于相应的时间点测相同的指标。结果经气管给家兔接种肺炎链球菌以后可以产生肺部感染,病理改变由轻到重表现为血管病变,急性支气管炎,局灶性支气管肺炎,实变以及肺脓肿等。大体标本观察有明显的充血及颜色变化等改变,细菌培养阳性。结论成功的建立了家兔肺炎链球菌肺炎实验模型