Spread of old and new clones of epidemic methicillin-resistant Staphylococcus aureus in Poland

Objective: To study the relatedness among methicillin-resistant Staphylococcus aureus (MRSA) isolates originating from two regions of Poland using different epidemiologic typing methods.
Methods: Forty-five MRSA isolates (19 from Warsaw and 26 from the Grajewo region) were collected between 1995 and 1996. For phenotypic epidemiologic analysis, antimicrobial susceptibility testing (AST) with a panel of 19 antibiotics was performed. For genotypic epidemiologic analysis, pulsed-field gel electrophoresis (PFGE) of Smal-digested chromosomal DNA, restriction endonuclease analysis of plasmid (REAP) DNA digested by Hin dIII, random amplification of polymorphic DNA (RAPD) and binary typing (BT) of genomic DNA by hybridization with five different RAPD-generated strain-specific DNA probes, were used.
Results: Six clusters of clonally related strains were found among the MRSA isolates analyzed. Three of these, identified in both regions, were related to previously described Polish epidemic clones, designated HeEMRSA-Pol1 (heterogeneously methicillin resistant—18 isolates) and HoEMRSA-Pol1 (homogeneously resistant—two clones, six isolates each). The remaining three clones, identified in the Grajewo region only, are previously undescribed. One of these, represented by 11 isolates, appears to be new epidemic heterogeneous MRSA clone (HeEMRSA-Pol2). Results of PFGE and BT in general showed good correlation, and, in some cases, RAPD using AP1 and AP7 primers could discriminate between isolates belonging to single PFGE or BT types. Broad AST and REAP can provide useful additional information concerning relatedness.
Conclusion: Evidence for the spread of previously recognized epidemic MRSA clones in Poland and the presence of a new epidemic heterogeneously resistant clone of MRSA in hospitals outside Warsaw is documented.     

应用IRS-PCR对金黄色葡萄球菌分型的研究

目的探讨低频限制性位点聚合酶链反应(infrequent-restriction-site PCR，IRS-PCR)在金黄色葡萄球菌(简称金葡菌)基因分型中的应用价值。方法建立本实验室IRS-PCR方法。同时用IRS-PCR和脉冲场凝胶电泳(PFGE)对金葡菌进行基因分型。根据49株社区感染分离菌的分型结果计算辨别力指数(ID)值估计分辨力。对其中30株社区感染菌重复实验一次估计重复性。比较两种基因分型方法的分型率、分辨力、重复性、结果的一致率及操作特点。结果建立IRS-PCR对金葡菌基因分型的方法。70株金葡菌均可被2种方法分型，分型率100％。IRS-PCR分为38个型，21株院内感染菌分为6个型，49株社区感染菌分为32个型，计算ID值为0．981。PFGE分为40个型，21株院内感染菌分为6个型，49株社区感染菌分为34个型，计算ID值为0．983。两种分型方法的重复性均为100％。对院内感染菌，两种方法分型的一致率为100％；对社区感染菌，两种方法分型的一致率为92％(45／49)。与PFGE相比，IRS-PCR更简单、省时、易于操作、不需特殊昂贵仪器。结论IRS-PCR能对金葡菌简易快速可靠分型，适合检验科对临床标本的快速有效分型，是一种有价值的分子流行病学研究工具。     

Antimicrobial susceptibility patterns of Staphylococcus aureus in Poland obtained by the National Quality Assurance Programme

As part of the Polish external quality assurance scheme, clinical laboratories were asked to send five consecutive isolates of Staphylococcus aureus and the corresponding susceptibility results to the national Centre of Quality Control in Microbiology. Of 1376 isolates submitted as S. aureus from 276 medical centres, 13 (< 1%) had been misidentified by local laboratories. Of 181 (13.5%) methicillin-resistant S. aureus (MRSA) isolates, most were identified correctly (c. 98% of laboratories). Although all MRSA isolates were fully susceptible to vancomycin, teicoplanin and linezolid, they were usually multiresistant; almost 23% were resistant to seven antimicrobial agents. Most (> 90%) MSSA isolates were susceptible to the tested antibiotics, except penicillin (21% susceptible) and tetracycline (62.4% susceptible). In addition to evaluating the proficiency of testing by local laboratories, the study yielded valuable information regarding the susceptibility patterns of S. aureus isolates in Poland.     

Whole organisms and purified cell walls compared as immunosorbents for the detection of IgE antibodies to Staphylococcus aureus

We have developed an immunoradiometric assay for IgE antibodies to Staphylococcus aureus (Staph IgE-Ab) which uses purified cell walls (PCW) from the Wood 46 strain of S. aureus as an immunosorbent. We compared Wood 46 PCW and whole organisms (WO) as immunosorbents for Staph IgE-Ab by performing tests on sera from patients with atopic dermatitis (AD) or the hyperimmunoglobulin E syndrome (hyper IgE syndrome). Sera with Staph IgE-Ab demonstrated dose-dependent binding to PCW and WO, but the ratio of specific to non-specific binding was much greater with PCW. Mean non-specific binding to WO was greater than to PCW, 5% versus 2%; and non-specific binding to WO varied directly with the serum concentration of IgE. Results of tests on patients' sera indicated that PCW are required in screening assays for Staph IgE-Ab to avoid false positive results caused by high levels of non-specific binding to WO.     

发病前细菌感染抑制过敏性哮喘发作的实验研究

目的： 研究发病前细菌感染对过敏性哮喘发病的影响。 方法： 用豚鼠作动物模型，观察金黄色葡萄球菌、大肠杆菌、链球菌、肺炎双球菌等4种呼吸道常见细菌感染之后哮喘发作时肺泡灌洗液中的白细胞总数、酸性粒细胞总数和百分比以及肺部的病理变化。 结果： 各细菌感染组肺泡灌洗液(BALF)中的白细胞总数均明显低于对照组，金黄色葡萄球菌组和大肠杆菌组酸性粒细胞总数和百分比明显降低，病理结果表明各细菌感染组的病变程度都要明显轻于对照组。 结论： 部分细菌感染后能够抑制哮喘的发作。     

Course and outcome of Staphylococcus aureus bacteraemia: a retrospective analysis of 308 episodes in a Swiss tertiary-care centre

Staphylococcus aureus bacteraemia (SAB) is associated with substantial morbidity and mortality worldwide. The charts of adult patients with SAB who were hospitalised in a Swiss tertiary-care centre between 1998 and 2002 were studied retrospectively. In total, 308 episodes of SAB were included: 2% were caused by methicillin-resistant strains; 49% were community-acquired; and 51% were nosocomial. Bacteraemia without focus was the most common type of community-acquired SAB (52%), whereas intravenous catheter-related infection predominated (61%) among nosocomial episodes of SAB. An infectious diseases (ID) specialist was consulted in 82% of all cases; 83% received appropriate antibiotic treatment within 24 h of obtaining blood cultures. Overall hospital-associated mortality was 20%. Community-acquired SAB was associated independently with a higher mortality rate than nosocomial SAB (26% vs. 13%; p 0.009). Independent risk-factors for a fatal outcome were age (p < 0.001), immunosuppression (p 0.007), alcoholism (p < 0.001), haemodialysis (p 0.03), acute renal failure (p < 0.001) and septic shock (p < 0.001). Consultation with an ID specialist was associated with a better outcome in univariate analysis (p < 0.001). Compared with a previous retrospective analysis performed at the same institution between 1980 and 1986, there was a 140% increase in community-acquired SAB, a 60% increase in catheter-related SAB, and a 14% reduction in mortality. In conclusion, mortality in patients with SAB remained high, despite effective antibiotic therapy. Patients with community-acquired SAB were twice as likely to die as patients with nosocomial SAB. Consultation with an ID specialist may reduce mortality in patients with SAB.     

临床分离4364株细菌的分布特征和耐药性变迁分析

目的了解2003—2005年分离的4364株细菌的分布特征及耐药性变迁，为合理使用抗菌药物提供依据。方法大多数分离细菌的鉴定和药敏试验利用BD Phoenix仪，少数利用手工鉴定和K-B法药敏试验。数据分析用WHONET5．0软件。结果葡萄球菌对万古霉素和替考拉宁的敏感率一直为100％，对其他抗菌药物的耐药率大多逐年上升，甲氧西林耐药率也逐年上升，金黄色葡萄球菌从40．8％上升到61．6％，表皮葡萄球菌从69．7％上升到79．8％。G^-杆菌合计，大多数抗菌药物的耐药率逐年升高，耐药率一直低于30％的为美洛培南、亚安培南、头孢哌酮／舒巴坦、哌拉西林-他唑巴坦和头孢他啶。大肠埃希菌和肺炎克雷伯菌体外产ESBLs的检出率一直居高不下，为29．5％～45．5％。结论本院肠杆菌科产ESBLs比例、葡萄球菌甲氧西林耐药率和非发酵菌碳青霉烯类耐药率均较高，应加强抗菌药物的合理使用和采取有效的隔离措施以降低耐药率及多重耐药菌的扩散。     

Organization of secondary lymphoid tissue and local IgE formation to Staphylococcus aureus enterotoxins in nasal polyp tissue

BACKGROUND: Bilateral nasal polyposis (NP) is characterized by high concentrations of IgE in NP tissue, which show no relation to the atopic status. We aimed to study the relationship between systemic and local IgE formation, nasal carriage of Staphylococcus aureus and nasal polyposis. METHODS: In serum and nasal tissue homogenates from 24 NP patients and 12 controls, we determined concentrations of total IgE and IgE antibodies to inhalant allergens and S. aureus enterotoxins (SAEs; A,B,C,D,E,TSST) by ImmunoCAP. Tissue cryosections were stained for CD3, CD20, CD38, CD23, FcepsilonRI, IgE and SEA/SEB. RESULTS: We demonstrated a higher incidence of S. aureus colonization (17/24) and IgE antibodies to SAEs in NP tissue (12/24) compared with controls (3/12 and 0/12, respectively). Total IgE and IgE antibodies in serum and NP tissue were dissociated because of local polyclonal IgE formation in NP tissue. Staining of NP tissue revealed follicular structures characterized by B and T cells, and lymphoid accumulations with diffuse plasma cell infiltration. CONCLUSIONS: We demonstrated the organization of secondary lymphoid tissue in polyp tissue and a polyclonal hyper-immunoglobulinemia E associated with the presence of IgE antibodies to SAEs, colonization with S. aureus, and tissue eosinophilia in a relevant subgroup of polyp patients.