ELISA法检测肠杆菌科细菌共同抗原在尿路感染快速诊断中的…

肠杆菌科细菌共同抗原（ＥＣＡ）是几乎存在于所有肠杆菌科细菌表面的氨基糖类抗原。本文介绍从Ｅ． ｃｏｌｉ Ｏ１４菌中提取、纯化ＥＣＡ，制备抗ＥＣＡ抗体，建立检测标本中ＥＣＡ的ＥＬＩＳＡ法。６９９例中段尿用本法与常规培养法比较，细菌数≥１０＾５／ｍｌ的２１１份标本中２０８例ＥＣＡ阳性，＜１０＾５／ｍｌ的８份标本中阳性２例。本法较培养法可提前１～２天发报告，对尿路感染有早期诊断价值。     

基于微流控芯片的实时荧光定量 ＰＣＲ 技术快速检测血小板制剂细菌 污染

摘要：目的 通过微流控芯片检测平台应用 Ｔａｑｍａｎ 探针的实时定量 ＰＣＲ 技术对血小板制剂中的细菌１６Ｓ ｒＤＮＡ 进行检测，探 讨该体系在血小板细菌污染的快速检测的应用。方法 向机采血小板中人为添加一定浓度的金黄色葡萄球菌或铜绿假单胞 菌，模拟成１０２ ～１０８ ＣＦＵ／ ｍＬ 细菌污染的血液标本，经１０倍梯度稀释后进行细菌计数及微流控芯片 ＦＱ ＰＣＲ 检测细菌１６Ｓ ｒＤＮＡ，并 对检测体系进行特异性、检出限和重复性评价。结果 微流控芯片 ＦＱ ＰＣＲ 体系特异性较强，探针和引物对阳性标本均有反 应，与空白对照无反应。对污染血小板的金黄色葡萄球菌，该方法的最低检出限为 ８６５ ＣＦＵ／ ｍＬ，而对铜绿假单胞菌的最低检 出限为 ８８５ ＣＦＵ／ ｍＬ。当金黄色葡萄球菌与铜绿假单胞菌浓度分别达到最低检测限时，空白对照与各细菌组的 Ｃｔ 值之差分别 为 ４．３５±１．０１ 和 ２．０３±０．６１。各浓度菌液提取的 ＤＮＡ 进行微流控芯片 ＦＱ ＰＣＲ 扩增后的 Ｃｔ 值计算重复性指数在 ０．０１２～ ０．０５２ 范围内。结论 微流控芯片 ＦＱ ＰＣＲ 检测平台可特异、有效地检出血小板中的细菌污染，为确保输血安全提供帮助。     

应用中消毒液细菌污染与使用情况调查

１９９６年 １月至 １９９９年１月,对全院门诊病房应用中的 ２％戊二醛、含 ６２５ ｍｇ／ Ｌ季铵盐的１２１０消毒液（以二癸基二甲基氯化铵及正烷基二甲基苄基氯化铵为主要成分的季铵盐复方消毒剂）、０．５％过氧乙酸、０．５％碘伏等消毒液,进行了细菌污染检测。检测时,对消毒液取样,加入相应中和剂中,混匀。中和作用 １０ｍｉｎ,检测细菌总数与致病菌。 检测３５８份消毒液,细菌总数超标（＞ １００ ｃｆｕ／ ｍｌ）率为１０．６％ （３８／３５８）。此外,有９份检出枯草芽胞杆菌,８份检出藤黄微球菌,７份检出产碱假单胞菌…     

2006/2008年革兰阴性杆菌对亚胺培南的耐药性调查

目的：分析革兰阴性杆菌对亚胺培南的耐药性变化。方法：对3a来从住院患者各种临床标本中分离的革兰阴性杆菌，进行回顾性分析。结果：3a共分离出5695株细菌，27个细菌种，肠杆菌科细菌3795株，占65．9％；非发酵菌1941株，占34．1％；分离率前3位的病原菌分别是大肠埃希菌、肺炎克雷伯菌、铜绿假单胞菌；2008年肠杆菌科细菌对亚胺培南的耐药率为0％；2008年嗜麦芽寡养单胞菌对亚胺培南的耐药率分别为100．0％。结论：亚胺培南对肠杆菌科细菌的体外抗菌活性强，醋酸钙不动杆菌和鲍氏不动杆菌对亚胺培南的耐药率呈上升趋势。     

超声波处理对评价戊二醛杀灭细菌芽胞效果的影响

为了解低浓度戊二醛对细菌芽胞的亚致死作用,采用悬液定量杀菌试验与超声波复苏的方法,检测了戊二醛对枯草杆菌黑色变种芽胞的杀灭效果。结果,戊二醛作用后,经超声波处理再接种培养,戊二醛对枯草杆菌黑色变种芽胞作用４～５ｈ的杀灭率较未经超声波处理者有所下降,其中１％戊二醛（即溶液中含戊二醛１０ｇ／Ｌ）更为明显。说明低浓度戊二醛对细菌芽胞有一定的亚致死作用。     

ICU内革兰氏阴性杆菌菌种分布及耐药性分析

目的：了解ICU内革兰氏阴性杆菌菌种分布情况及耐药性。方法：对ICU病房2004年2月-2005年4月所有临床标本分离的革兰氏阴性杆菌进行细菌鉴定和药敏检测。结果：ICU内革兰氏阴性杆菌以为铜绿假单胞菌为主，占37％。其次为肺炎克雷伯菌，占23％。嗜麦芽窄食菌占13％，不动杆菌占11％。产ESBLs肠杆菌科细菌占革兰氏阴性杆菌的29％。占肠杆菌科总数的85％。铜绿假单胞菌对头孢他定的耐药率为69％，对头孢哌酮／舒巴坦的耐药率为81％，对亚安培南的耐药率为57％，肺炎克雷伯菌对头抱哌酮／舒巴坦的耐药率为60％，对亚安培南的耐药率为0％，不动杆菌对头抱哌酮／舒巴坦的耐药率为27％。对亚安培南的耐药率为7．1％。嗜麦芽窄食菌对头孢哌酮／舒巴坦的耐药率为88％，ESBLs肠杆菌科细菌对亚安培南的耐药率为0％。结论：ICU内革兰阴性菌耐药性较高，产ESBLs情况严重，临床医生应尽量在确立了病原学诊断后参考细菌药敏报告用药，以减少耐药菌株的产生。     

2643株临床分离革兰阴性杆菌耐药性分析

目的对临床分离的革兰阴性杆菌的分布及耐药性进行分析,为临床合理应用抗生素提供依据。方法用Walkaway40型全自动细菌鉴定仪,对临床分离革兰阴性杆菌进行来源监测及耐药试验以及统计学分析。结果2643株革兰阴性杆菌中分离率居前五位的依次为大肠埃希菌（28．5％）、铜绿假单胞菌（17．3％）、肺炎克雷伯菌（14．7％）、阴沟肠杆菌（9．4％）、鲍曼不动杆菌（7．5％）。不同来源的标本病原菌分布不同：呼吸道标本以铜绿假单胞菌为主,其余标本以大肠埃希菌为主。药敏结果显示,肠杆菌科对头孢哌酮／舒巴坦和亚胺培南耐药率低于10．5％,对氨苄西林、美洛西林、复方新诺明、庆大霉素、头孢曲松、头孢噻肟耐药率达49．4％～97．4％。铜绿假单胞菌和鲍曼不动杆菌对头孢哌酮／舒巴坦和亚胺培南耐药率低于12．7％,对氨苄西林、美洛西林、复方新诺明、庆大霉素、头孢噻肟、头孢曲松、丁胺卡那、头孢他啶耐药率达50．2％～90．2％。结论大肠埃希菌、铜绿假单胞菌、肺炎克雷伯菌、阴沟肠杆菌、鲍曼不动杆菌是临床分离的主要革兰阴性杆菌,而且耐药情况严重,应加强耐药性监测,合理使用抗生素,以减少耐药菌株的传播流行。     

全自动细菌分析系统在检测产超广谱β-内酰胺酶细菌中的临床应用

目的　了解超广谱β内酰胺酶（ Ｅ Ｓ Ｂ Ｌｓ） 细菌在医院分离及分布情况,以利于对 Ｅ Ｓ Ｂ Ｌｓ 细菌的监控和治疗。方法　用 Ｖ Ｉ Ｔ Ｅ Ｋ３２ 型全自动细菌分析系统药敏检测卡 Ｇ Ｎ Ｓ Ｎ Ｔ 进行细菌 Ｅ Ｓ Ｂ Ｌｓ 的测定。结果　在１３２ 株大肠埃希菌、１１９ 株肺炎克雷伯菌、６０ 株阴沟肠杆菌中 Ｅ Ｓ Ｂ Ｌｓ 的检出分别为２８８ ％ 、１９３ ％ 、５０ ％ ,而７３ 株绿脓假单胞菌未检到 Ｅ Ｓ Ｂ Ｌｓ 菌;各病区产 Ｅ Ｓ Ｂ Ｌｓ 菌的分离率以重症监护（ Ｉ Ｃ Ｕ） 病房最高（３３６ ％ ） ,其次为慢性病病房和干部病房（ 各为２００ ％ ） ,介入科病房（１９６ ％ ） ,呼吸科病房（１２９ ％ ） ,其他病房（３０ ％ ） ;产 Ｅ Ｓ Ｂ Ｌｓ 菌对亚胺培南全部敏感,对头孢哌酮／ 舒巴坦未发现有高耐株,中介占２３４ ％ （１５／６４） 。结论　 Ｖ Ｉ Ｔ Ｅ Ｋ３２ 型全自动细菌分析系统的 Ｇ Ｎ Ｓ Ｎ Ｔ 卡可正确检测产 Ｅ Ｓ Ｂ Ｌｓ 菌, Ｉ Ｃ Ｕ 病房是产 Ｅ Ｓ Ｂ Ｌｓ 菌的主要来源,对产 Ｅ Ｓ Ｂ Ｌｓ 菌的治疗以亚胺培南最佳,其次为高浓度的头孢哌酮／ 舒巴坦。     

人凝血因子Ⅸ∶Ag酶联免疫测定法的建立

采用抗人凝血因子Ⅸ单克隆和多克隆两种抗体，建立了人凝血因子Ⅸ抗原（ＦⅨ∶Ａｇ）酶联免疫测定法（ＥＬＩＳＡ）。标准曲线相关系数为０．９９３（Ｐ＝０．００１，ｎ＝５），测定范围（６．２５～１００）Ｕ／Ｌ，最低检测量１．５Ｕ／Ｌ；批内及批间变异系数分别为７．２５％和４．７３％，回收率９９．７％；与人血浆中其它蛋白质未见交叉反应。测得２０人份正常人血浆中ＦⅨ∶Ａｇ为（１０２７±１７２）Ｕ／Ｌ，与ＦⅨ促凝活性（ＦⅨ∶Ｃ）测得值比较，差异无显著性（Ｐ＞０．５）。重症乙型血友病人血浆ＦⅨ∶Ａｇ测得值与临床诊断相符。     

聚合酶链反应检测耐甲氧西林金黄色葡萄球菌mecA基因

建立聚合酶链反应（ｐｏｌｙｍｅｒａｓｅｃｈａｉｎｒｅａｃｔｉｏｎ，ＰＣＲ）检测耐甲氧西林金黄色葡萄球菌（ｍｅｔｈｉｃｉｌｌｉｎ－ｒｅｓｉｓｔａｎｔＳ．ａｕｒｅｕｓ，ＭＲＳＡ）ｍｅｃＡ基因的技术。四种ＤＮＡ提取方法检测灵敏度依次为超声裂解法（５×１０５ＣＦＵ／ｍｌ）、溶菌酶表面活性剂法（５×１０６ＣＦＵ／ｍｌ），表面活性剂法（１×１０７ＣＦＵ／ｍｌ）和直接煮沸法（１×１０８ＣＦＵ／ｍｌ），直接煮沸法具有简便性。引物的正链位于１８１～２００、负链位于３１１～３３０，序列分别为５＇－ＧＡＡＡＴＧＡＣＴＧＡＡＣＧＴＣＣＧＡＴ，５＇－ＧＣＧＡＴＣＡＡＴＧＴＴＡＣＣＧＴＡＧＴ，其扩增产物长度为１５０ｂｐ。五种常见菌证明本法具有较高特异性。苯唑青霉素ＭＩＣ水平与ｍｅｃＡ基因之间具有很好的相关性，ＭＩＣ≥４μｇ／ｍｌ的２２株菌均检出ｍｅｃＡ基因，提示苯唑青霉素常规检测ＭＲＳＡ的可行性。ＰＣＲ方法对于隐匿型耐药株的检出具有重要价值。     

老年人尿路感染常见病原菌耐药性分析

目的研究老年人尿路感染的常见病原菌及其对常用抗生素的耐药性，以指导临床用药。方法采用Vitek-Ams System和Kirby-Bauer法对267例尿路感染的老年患者标本进行细菌培养并检测其对多种抗生素的耐药性。结果共检出病原菌310株，其中革兰阴性杆菌201株（66．1％），主要为大肠埃希菌137株（45．1％）、铜绿假单胞菌25株（8．2％）、肺炎克雷伯菌19株（6．3％）、变形杆菌13株（4．3％）。革兰阴性杆菌对第3代头孢菌素的耐药率较高，可能与产超广谱B内酰胺酶菌株的增多有关，亚胺培南是治疗革兰阴性杆菌感染最有效的药物。革兰阳性球菌76株（23．0％），层析万古霉素仍然是治疗革兰阳性球菌感染最有效的药物。真菌33株（10．9％），对两性霉素B耐药率为0。结论老年人尿路感染病原菌耐药性严重，持续耐药监测对抗生素的合理应用，提高疗效，减缓耐药菌株的发生与发展有重要意义。     

Treatment of experimental endocarditis caused by a beta-lactamase-producing strain of Enterococcus faecalis with high-level resistance to gentamicin.

Several antimicrobial regimens were evaluated in the treatment of experimental enterococcal endocarditis due to a beta-lactamase-producing, highly gentamicin-resistant strain of Enterococcus faecalis. Ampicillin alone cleared bacteremia in the majority of rats and reduced titers of bacteria within vegetations (6.84 versus 8.80 log10 CFU/g in controls) but did not sterilize valves. Ampicillin-sulbactam combinations, vancomycin, daptomycin, and imipenem each reduced residual bacterial titers within vegetations to 4.01 log10 CFU/g or less; in 26 to 43% of animals receiving 5 days of therapy, titers of bacteria were reduced to undetectable levels. In a separate experiment, rats received ampicillin-sulbactam, daptomycin, or vancomycin for 10 days and were then observed for 10 days after termination of therapy for evidence of relapse. In surviving rats, valves remained sterile in four of five rats treated with ampicillin-sulbactam, in five of seven treated with daptomycin, but in only one of eight receiving vancomycin.     

淋病奈瑟菌PCR检验质量控制结果分析

临床分离的淋病奈瑟菌以无菌生理盐水１０倍系列稀释，制备成不同浓度的菌悬液，作为ＰＣＲ质控 发给广东省部分开展淋病ＰＣＲ检验的实验室，２３个回报结果中，假阳经１７．６％。１０ＣＦＵ淋球菌检出率１５．７％，１０＾２ＣＦＵ检出率２７．２％，１０＾３ＣＦＵ检出率５３．９％。总符合率４４．８％，结果表明，一些实验室不仅存在假阳性问题，而且阳性检出率也不高，应引起主度重视。     

Decontamination effect of milling by a jet mill on bacteria in rice flour

The decontamination effect of milling by a jet mill was investigated by counting the number of bacteria in brown and white rice flour with mean particle diameters of 3, 20, and 40μm prepared by the jet mill. In the jet mill, the particles are crushed and reduced in size by the mechanical impact caused by their collision. Although the brown and white rice grains were contaminated with approximately 10(6) and 10(5) CFU/g bacteria, the microbial load of the rice flour decreased as the mean particle diameter decreased, ultimately decreasing to approximately 104 and 103 CFU/g in the brown and white rice flour. The temperature and pressure changes of the sample were not considered to have an effect on reducing the bacterial count during the milling. Hence, it was thought that the rice flour was decontaminated by other effects.     

Pharmacokinetics and efficacies of liposomal and conventional formulations of tobramycin after intratracheal administration in rats with pulmonary Burkholderia cepacia infection

The objective of the present study was to determine the pharmacokinetics and efficacies of liposomal and conventional formulations of tobramycin against Burkholderia cepacia in a model of chronic lung infection. Male Sprague-Dawley rats were inoculated intratracheally with 10(6) CFU of a very resistant strain of B. cepacia (strain BC 1368; MIC, 128 micro g/ml) to establish lung infection. A 1,200- micro g dose of tobramycin was administered intratracheally as a liposomal formulation and as a conventional formulation. Rats were anesthetized and exsanguinated by cardiac puncture at different times over 24 h to assess pulmonary tobramycin concentrations and the number of residual CFU. Pharmacokinetic parameters were calculated by using a two-compartment model with NONMEM. The mean half-life at the beta phase (t(1/2beta)) and the pulmonary exposure (the area under the concentration-time curve [AUC]) of liposomal tobramycin were 19.7 h (coefficient of variation [CV], 24.2%) and 6,811 micro g. h/lungs (CV, 19.7%), respectively. The pharmacokinetics of conventional tobramycin were statistically different, with a t(1/2beta) and AUC of 12.9 h (CV, 31.4%) and 821 micro g. h/lungs (CV, 15.0%), respectively. Pearson chi-square analyses were performed on residual CFU data distributed in the following categories: <10(3), 10(3) to 10(5), and >10(5). Differences in CFU data between formulations showed a statistical trend (P < 0.10) when data from all time points were used, and statistically significant differences were found after 12 h (P < 0.05), with greater eradication achieved with the liposomal formulation. In conclusion, intratracheal administration of tobramycin in liposomes was associated with marked changes in the pharmacokinetics of the drug in the lung and an apparent trend for a prolonged efficacy against B. cepacia. These results support the hypothesis that inhalation of liposomal tobramycin may improve the management of chronic pulmonary infections caused by resistant bacteria in patients with cystic fibrosis.     

改良麦康凯平板筛查肠道定植耐碳青霉烯类肠杆菌科细菌的性能评价

目的建立一种运用改良麦康凯平板快速筛查肠道定植耐碳青霉烯类肠杆菌科细菌(CRE)的方法,并进行性能评价。方法分别制备含有终浓度为4μg/mL亚胺培南、4μg/mL美罗培南、2μg/mL厄他培南的改良麦康凯平板。选用临床连续分离45株CRE及30株非CRE菌株,检测改良麦康凯平板的敏感性及特异性,采用其中20株CRE菌株对改良平板的最低检测限进行评估。PCR方法检测CRE菌株耐药基因(TEM、SHV、NDM、KPC、OXA-48、VIM、IMP)。收集142例粪便样本评价改良平板的筛查性能,并与肉汤增菌法及CHROMagar KPC显色培养基进行比较。结果45株CRE菌株中TEM、SHV检出率分别为62.22%(28/45)和24.44%(11/45),NDM、KPC、OXA-48的检出率分别为62.22%(28/45)、15.56%(7/45)和13.33%(6/45),未检出VIM和IMP。45株CRE菌株在厄他培南改良平板上均生长,亚胺培南改良平板生长44株,美罗培南改良平板生长43株,敏感性分别为100%(45/45)、97.78%(44/45)和95.56%(43/45)。30株非CRE菌株在亚胺培南与美罗培南改良平板上均未生长,在厄他培南改良平板上生长2株,特异性分别为100%(30/30)、100%(30/30)和93.33%(28/30)。20株CRE中,接种菌液浓度为10⁵~10⁸ CFU/mL时,亚胺培南、美罗培南和厄他培南改良平板的检出率分别为70%(14/20)、65%(13/20)和100%(20/20);接种浓度为10⁵ CFU/mL以下时亚胺培南、美罗培南、厄他培南改良平板的检出率分别为10%(2/20)、10%(2/20)和95%(19/20)。亚胺培南、美罗培南改良麦康凯平板最低检测限大多分布在10⁵~10⁸ CFU/mL接种水平,而厄他培南改良麦康凯平板最低检测限集中分布于10²~10³ CFU/mL。142份粪便样本中,亚胺培南、美罗培南、厄他培南改良麦康凯平板分别筛查出3株、2株、4株CRE,均未检出非CRE菌株;肉汤增菌法检出5株CRE及2株非CRE;CHROMagar KPC显色培养基检出5株CRE及1株非CRE。3种改良平板与肉汤增菌法及CHROMagar KPC显色培养基筛查结果间差异均无统计学意义(P>0.05)。与亚胺培南和美罗培南相比,厄他培南平板筛查结果与肉汤增菌法、CHROMagar KPC显色培养基一致性更好,Kappa值分别为0.717和0.885。结论改良麦康凯平板筛查肠道CRE菌株敏感性和特异性高,且操作简便,价格低廉,对操作人员技术要求低。厄他培南改良麦康凯平板具有更低的检测限,且与肉汤增菌法及CHROMagar KPC显色培养基筛查结果有更好的一致性,更适合推广使用。     

Best practice for patient self-cleaning of urinary drainage bags

The best practice for cleansing urinary drainage bags (leg and bed) in a hospital setting can be translated for use by clients for self-care within the home setting. Common household bleach containing 5.25% sodium hypochlorite, diluted in a 1:10 ratio with tap water, is a safe and effective method to reduce colony-forming units per milliliter (CFU/ml) to a study-defined acceptable range of 100 CFU/ml when a standard procedure was consistently used. This practice increases the life of the urinary drainage bag, decreasing the cost for the client. Ascending bacteria from the urinary drainage bag may also decrease, resulting in fewer urinary tract infections.     

Floating pellets containing bacterial antagonist for control sheath blight of rice: formulations, viability and bacterial release studies.

Floating pellets containing spores of bacterial biological control agent, Bacillus megaterium were prepared by extrusion-spheronization process. The formulations composed of hydrogenated vegetable oil (HVO), lactose, microcrystalline cellulose (Avicel(R) PH101), and a disintegrant; cross-linked sodium carboxymethylcellulose (Ac-Di-Sol(R)). The finishing pellets contained bacteria ranging from 10(7) to 10(8) CFU/g and the viability of bacteria in all formulations remained high after 6 months storage. The scanning electron microscope (SEM) was used to observe endospores of B. megaterium on both the surface and the inside of the pellets. The formulations were tested for their physical properties, floating ability and bacterial release. The level of disintegrant in the formulations influenced the floating ability and the liberation of antagonistic bacteria from pellets. The bacterial pellets showed promising result in suppression of the development of sheath blight lesions in greenhouse experiment.     

全自动尿沉渣分析仪UF-100中细菌计数参数阈值的研究

目的：研究尿分析仪UF-100中细菌计数参数的阈值。方法：以462份住院患者清洁中段尿液为标本,以细菌定量培养为对照,建立ROC曲线确定阈值;通过拟合曲线确定细菌计数和定量培养结果相互关系。结果：以培养结果102CFU/ml、104CFU/ml和105CFU/ml为阈值,细菌计数阈值分别是5459/μl、6520/μl和8092/μl。不同性别、不同菌种阈值有不同。培养计数对数值（Y）-细菌计数值（X）的直线回归是Y=1.9302＋0.0002X,对数回归是Y=-6.1412＋1.1214×lnX。结论：定量培养确定UF-100中细菌计数参数的阈值,该值随性别、菌种不同而异。通过回归曲线及细菌计数结果可以预测定量培养结果。     

Penetration of cefixime into fibrin clots and in vivo efficacy against Escherichia coli, Klebsiella pneumoniae, and Staphylococcus aureus.

The experimental model of infected fibrin clots in rabbits was used to study the penetration and in vivo activity of cefixime against Klebsiella pneumoniae, Escherichia coli, and Staphylococcus aureus. The respective MICs of cefixime against these strains were 0.25, 2, and 8 micrograms/ml. The clots were infected with 10(6) to 10(8) CFU/g. Groups of four animals for each strain received an intravenous injection of 100 mg of cefixime per kg over 30 min. High peak levels were observed in serum (146.5 micrograms/ml) and clots (15.8 micrograms/g), and the antibiotic was still detectable in the clots (0.6 micrograms/g) 24 h after administration. The respective serum and clot elimination half-lives were 0.7 and 5.0 h. The mean serum protein binding was 23.8 +/- 3.8%. Cefixime was highly bactericidal against K. pneumoniae and E. coli and reduced, over a 24-h period, their respective colony counts by 7.8 log10 and 6.2 log10 CFU/g of fibrin. It was less effective against S. aureus but still reduced the bacterial counts by 2.8 log10 CFU/g of fibrin. The present results demonstrate that cefixime, a new broad-spectrum oral cephalosporin, has a long tissue half-life which ensured, at the dose given here, good in vivo bactericidal activity against both gram-positive and gram-negative bacteria up to 24 h after administration of the antibiotic.