临床常见葡萄球菌蛋白指纹图谱的建立

目的 建立临床常见三种葡萄球菌的蛋白指纹图谱,为其快速诊断奠定基础.方法 收集临床常见葡萄球菌包括金黄色葡萄球菌、表皮葡萄球菌及溶血性葡萄球菌,利用16S rDNA测序技术对其进行鉴定;表面增强激光解吸电离飞行时间质谱(SELDI-TOF MS)技术检测其蛋白表达,采用Ciphergen Protein chip软件自动采集数据.评价该检测方法的重复性及细菌蛋白表达的稳定性.结果 在分子量3000 ～20000 Da范围内,3种葡萄球菌有各自特异的蛋白指纹图谱;SELDI-TOF MS检测细菌蛋白的孔间重复性和芯片间重复性好,同一株细菌4次重复的蛋白指纹图谱相似;同种细菌不同株之间蛋白指纹图谱表达稳定,共有蛋白峰分子量变异系数≤0.5％.结论 在分子量3000 ～20000 Da范围内,各细菌蛋白指纹图谱差异明显,为细菌的快速鉴定提供了新方法.     

生殖系统的菌群分布和药敏分析

对我院１９９６～１９９７年度性病门诊、皮肤科门诊送检的生殖道标本（不包括淋菌培养）,按照《全国临床检验操作规程》进行分离培养,分离出革兰氏阳性球菌９１例,革兰氏阴性杆菌２５例,用美国ＢＤ公司Ｓｃｅｐｔｏｒ半自动细菌鉴定仪作鉴定和药敏试验（ＭＩＣ法）。凝固酶阴性葡萄球菌引起的感染最多,占５１％,其次为大肠埃希氏菌（１６％）,粪肠球菌（１２％）,金黄色葡萄球菌（９％）,无乳链球菌（６％）,肺炎克雷伯氏菌（３％）,奇异变形杆菌（３％）。药敏结果显示,革兰氏阳性球菌对万古霉素、氯洁霉素有良好敏感性,革兰氏阴性杆菌对泰能、丁胺卡那霉素敏感性高。     

961份血培养的细菌及其药敏分析

对９６１份血培养的细菌和药敏结果进行了分析。结果：有１２３份培养阳性,阳性率为１２．８％,共分离出１２６株细菌。其中革兰氏阳性球菌９１（Ｇ~＋Ｃ）株,占７２．２％,以表皮葡萄球菌．金黄色葡萄球菌和粪肠球菌为主,占所有检出菌的５６．３％;革兰氏阴性杆菌（Ｇ~－Ｂ）２６株,真菌７株,提示革兰氏阳性球菌为本地区引起菌血症的主要病原菌,真菌感染有增多趋势。药敏结果提示：针对Ｇ~＋Ｃ的首选药物为万古霉素,对其它抗菌素都有普遍的不同程度的耐药性,应引起临床注意。     

大肠埃希菌蛋白指纹图谱的建立

目的建立大肠埃希菌（Escherichia coli,E．coli）蛋白指纹图谱,为Ecoli感染快速诊断奠定基础。方法收集临床分离E.coli88株,提取细菌DNA,PCR检测Ecoli 16S rRNA。蛋白提取液提取细菌蛋白,干化学法测蛋白浓度,应用表面增强激光解析电离飞行时间质谱技术（SELDI-TOF-MS）检测Ecoli蛋白,采用Ciphergen Pro-teinchip软件自动采集数据。重复测定20次Ecoli混合标本,评价SELDI检测Ecoli蛋白分子量的重复性。结果E．coil标准菌株ATCC 25922和临床分离株均可检出16S rRNA。AU芯片能捕获近30个E．coli蛋白峰,其中19个蛋白峰构成E．coli特征性蛋白指纹图谱,各蛋白峰在临床分离E．coli间分子量变异系数≤0．2％。SELDI重复检测20次E．coli混合标本显示同一蛋白峰的分子量变异系数≤0．05％。结论E．coli在分子量3～20kD范围内具有特征性蛋白指纹图谱,为快速诊断E.coli感染提供了新思路。     

细菌菌群的分布及耐药性分析

目的：了解本院临床分离致病菌菌群的分布及耐药情况,给临床经验用药提供可靠依据。方法：按《全国临床微生物检验操作规程》培养分离菌种,用美国BD公司的Sceptor半自动细菌鉴定仪或法国梅里埃公司的Vitek-60细菌鉴定仪进行鉴定及药物敏感实验,结果：临床分离 的致病菌中革兰阳性球菌占39．6％,革兰阴性杆菌占60．4％,前5位细菌分别为铜绿假单胞菌188株（17．0％）,大肠埃希菌143株（13．0％）,表皮葡萄球菌140株（12．7％）,肠球菌115株（10．7％）,金黄色葡萄球菌109株（9．9％）,药敏结果显示,苯唑西林耐药葡萄球菌（MRS）的耐药率明显高于苯唑西林敏感葡萄球菌109株（9．9％）,药敏结果显示,苯唑西林耐药葡萄球菌（MRS）的耐药率明显高于苯唑西林敏感葡萄球菌（MSS）,球菌中未分离出万古霉素耐药的菌株,但肠球菌中检测出有对万古霉素中介的菌株;肠杆菌科细菌对亚胺培南的耐药率最低,铜绿假单胞菌对头孢他啶,头孢哌酮,氨曲南,丁胺卡那霉素,亚胺培南的耐药率亦较低。结论：革兰阴性杆菌分离率高于革兰阳性球菌;革兰阳性球菌对万古霉素敏感率最高,肠杆菌科对亚胺培南敏感率最高,非发酵菌对亚胺培南,氨基糖甙类,头孢他啶,头孢哌酮药蓄亦较低。     

基于通用探针库的实时荧光PCR技术快速检测阳性血培养瓶中的细菌病原体

为从阳性血培养瓶中快速检测8种细菌,选取Roche公司通用探针库的锁核酸探针进行基于实时荧光PCR技术的一组试验。为鉴定菌种根据革兰氏染色的结果,对四组靶向于一个革兰氏阳性菌和一个革兰氏阴性菌的双重实时荧光PCR反应进行优化。检测的灵敏度是1-10CFU(菌落形成单位)/PCR反应混合物。试验的特异性和交叉反应由28个来自于ATCC(美国模式培养物集存库)的参考菌株和77种血培养阴性的标本进行鉴定。在这些试验样品中没有观察到交叉反应,因此证明试验100%的特异性。72种此前鉴定过的临床病毒株通过实时荧光PCR技术进行测试,验证实时荧光PCR试验的精确度和可行性。此外,用实时荧光PCR试验55类阳性血培养样本,通过生化分析评价,50(90.9%)类被鉴定为相同物种。实时荧光PCR与传统的方法有98.2%的一致。实时荧光PCR可以用来作为从阳性血培养中早期检测常见病原体方法的补充。     

VITEK2 Compact全自动微生物分析系统对牛乳中葡萄球菌的鉴定效果评价

【目的】评价VITEK2 Compact全自动微生物分析系统对牛乳中葡萄球菌属细菌鉴定效果。【方法】对从西宁、白银、西安三地乳样中分离的52株葡萄球菌通过VITEK2 Compact全自动微生物分析系统进行鉴定,并与革兰氏染色、触酶试验、血浆凝固酶试验以及16S r RNA基因序列分析结果进行比对;通过对凝固酶阴性葡萄球菌和凝固酶阳性葡萄球菌在不同麦氏浓度的鉴定,评价该系统的稳定性。【结果】VITEK2 Compact全自动微生物分析系统对葡萄球菌在"属"水平的鉴定结果符合率达96.15%,与血浆凝固酶结果对比,凝固酶阴性葡萄球菌属细菌鉴定结果符合率为92.86%,对凝固酶阳性葡萄球菌属细菌鉴定结果符合率为50%,但对具体种鉴定时,结果符合率仅为44.23%。在稳定性检测的试验中,鉴定符合率为100%,鉴定结果可靠性不随菌液浓度变化而变化。【结论】该系统可用于牛乳中葡萄球菌在"属"水平的鉴定。     

API系统鉴定化妆品及一次性卫生用品微生物种类的研究

利用API2 0E系统鉴定 18株肠杆菌科细菌 ,鉴定结果符合率 10 0 % ,并用API2 0E ,API 2 0NE ,APISTAPH和API2 0cAUX分别对分离自化妆品和一次性使用卫生用品的 183株革兰氏阴性发酵杆菌 ,革兰氏阴性非发酵杆菌 ,革兰氏阳性球菌和酵母菌成功进行了菌种鉴定 ,另有 3株革兰氏阴性氧化酶阴性杆菌未能鉴定到种     

MADLI-TOF MS自建库在马尔尼菲篮状菌快速鉴定中的应用

目的评价基质辅助激光解吸电离飞行时间质谱(MADLI-TOF MS)自建库对临床分离马尔尼菲篮状菌快速鉴定的能力。方法收集广州、玉林、防城港3个地区临床分离的马尔尼菲篮状菌,通过ITS区扩增测序进行菌种准确鉴定。菌株通过微型玻璃珠研磨结合甲酸/乙腈提取法预处理。通过采集建库菌株不同时间点、不同平板上菌丝相和酵母相菌落的质谱数据,建立包含马尔尼菲篮状菌超级图谱的自建库;并用非建库的马尔尼菲篮状菌及临床常见的真菌评估建立的自建库。结果建立了包含具有39个特征峰的马尔尼菲篮状菌超级图谱的自建库;用于评估的菌株均被鉴定到种,准确率为100%。结论 MADLI-TOF MS建立的马尔尼菲篮状菌自建库能够对于临床马尔尼菲篮状菌进行快速、准确的鉴定。     

Vitek-AMS鉴定临床细菌的应用研究

目的探讨Vitek-AMS对临床细菌鉴定的应用价值。方法对玉溪市人民医院1999年至2008年临床分离11 537株细菌(临床株)和省、部级临床检验中心下发微生物学室间质量评价鉴定菌种(参考株)的Vitek-AMS鉴定结果作对比分析。结果 11 537株临床分离菌中,不能鉴定细菌8株(0.07%);除外传染病因子,鉴定到属细菌114株(1.62%),鉴定到种(含亚种、物生型)细菌6 908株(98.38%)。共有64属193种。细菌类型的分布革兰阴性杆菌革兰阳性球菌酵母菌革兰阳性杆菌厌氧菌革兰阴性球菌。用参考菌种作比较,Vitek鉴定种的符合率为83.08%(54/65),属的符合率为98.46%(64/65);其中革兰阴性杆菌符合率最高(100%,24/24),酵母菌类符合率最低(66.67%,14/21)。7种鉴定卡菌种的阳性检出率平均为56.80%(234/412),其中GPI卡最高(100%),NHI卡最低(13.33%);但应用机会最多是GNI+卡。Vitek-AMS检测葡萄球菌产β-内酰胺酶阳性率为88.89%,大肠埃希菌和肺炎克雷伯菌产ESBLs阳性率分别为59.74%和32.20%。结论 Vitek-AMS的应用为临床细菌学检验提供了一种高效、快速、可靠的实验方法;但对于个别菌种、细菌酶的检测必要时应以参考方法确认。     

外科血培养标本中病原菌分布及耐药性分析

目的了解中山大学附属第一医院外科血标本中病原菌的菌种分布及常见菌株的耐药性。方法血标本用Bact/A lert-120全自动血培养仪进行血培养,阳性血培养转种后用VITEK-60 AMS细菌鉴定仪鉴定,用K-B法进行药敏试验。结果2002年1月至2005年12月血培养标本中共分离出病原菌256株,阳性率为10.6%。122株(47.7%)为革兰阴性杆菌,其中肠杆菌科细菌占72.1%(88/122),非发酵菌占27.9%(34/122);113株(44.1%)为革兰阳性球菌,其中葡萄球菌属占51.3%(58/113),肠球菌占38.1%(43/113);真菌21株(8.2%)。血培养中的革兰阴性杆菌对亚胺培南、美洛培南治疗敏感;革兰阳性球菌对万古霉素和替考拉宁敏感。结论肠杆菌科细菌和葡萄球菌是外科血培养中的主要病原菌,其耐药现象严重,宜根据药敏结果选用敏感抗菌药物治疗。     

The establishment of resistance phenotypes for bacteria isolated from outpatients in urine cultures

From 1911 outpatients, who addressed a Timi?oara private clinical laboratory, from January to December 2005, we collected 1,889 urine cultures, 431 being positive. Bacteria identification was generally done using morphological, cultural, biochemical characters and pathogenicity tests. Sensitivity testing to antimicrobial medical drugs was done by using the classical diffusion Kirby-Bauer method and the automatic analyzer Osiris, also. The main bacteria involved in the etiology of these infections were represented by Enterobacteriaceae, head of the list being Escherichia coli (81.21%), followed by Klebsiella pneumoniae (8.35%) and Proteus mirabilis (3.02%). We also isolated Gram positive cocci (in a much smaller proportion), mainly represented by Enterococcus faecalis (1.16%), Staphylococcus aureus (0.93%), Streptococcus agalactiae, and also Gram negative non-fermentative bacilli, such as Pseudomonas aeruginosa (0.93%) or Acinetobacter baumanii (0.23%). As soon as we performed the sensitivity tests, we divided them in resistance phenotypes: Most of the Enterobacteriaceae were integrated in the wild phenotype, followed by the penicillinase producing phenotype. An E. coli strain (0.29%) and 3 Klebsiella pneumoniae strains (8.33%) were integrated in the large spectrum, multidrug resistant, beta-lactamase producing phenotype, also associated with resistance to fluoroquinolones and aminoglycosides; Non-fermentative bacilli did not present special resistance problems, the four Pseudomonas aeruginosa strains were integrated in the wild phenotype (secreting induced chromosomal cephalosporinase). As for Staphylococcus aureus it was identified a strain having fluoroquinolone resistance, two strains secreting penicillinase and having a K (Nm) phenotype and a strain secreting penicillinase only. Antibiotic resistance represents a major concern for patients, physicians, healthcare managers, and policymakers. The use of antibiotics is closely linked with the development of acquired antibiotic resistance.     

晚期肿瘤患者下呼吸道感染临床分析

目的：探讨晚期肿瘤患者下呼吸道感染的细菌学分类情况及抗生素耐药性特点。方法：采用回顾性调查107例晚期肿瘤患者并下呼吸道感染痰菌分离的病原菌并分析其特点,药敏试验采用K—B纸片法。结果：137株病原菌,G杆菌67株(48．9％)．G^ 球菌44株(32．1%),真菌26株(19．0％)。主要致病细菌依次为铜绿假单胞菌19．7％．金黄色葡萄球菌13．9％,肺炎克雷伯菌10．9％。万古霉素、新霉素、环丙沙星对G^ 球菌较敏感;环丙沙星、阿米卡星、头孢哌酮对G杆菌作用较强．其他抗菌药物对各种病原菌均呈现不同程度的耐药。结论：晚期恶性肿瘤并下呼吸道感染的病原菌多为条件致病菌,耐药率高。应加强病原菌培养及药敏监测,以指导临床科学使用抗生素。     

临床病原菌种类及耐药性监测

目的 探讨病原菌种类及其对抗菌药物的耐的耐药状况。方法 收集１９９８年１月－１９９９年１２月临床感染标本分离的病原菌并分析其种类,药敏试验采用Ｋｉｒｂｙ－Ｂａｕｅｒ纸片扩散法。结果 １１８２株病原菌,革兰氏阳性球菌６０４株（５１．１％）,革兰氏阴性杆菌５７８株（４８．９％）。病原菌以金黄一萄球菌、大肠埃希菌、表皮葡萄球菌、铜绿假单胞菌最多见。去甲万古霉素、阿米卡星、新霉素对革兰氏阳性球菌抗菌作用较     

Biodiversity of the microbial flora associated with two strains of Culexpipiens (Diptera: Culicidae)

Bacterial Flora from external surface and alimentary canal of wild and laboratory strains of Culex pipiens were isolated and investigated using quantitative bacterial cultures. Individual colonies were subcultured and identified to species level. Counts from alimentary canal differ significantly from those of the external surface. An increase in bacterial density was detected after feeding on mammalian and avian blood meal. Bacterial identification revealed a complex bacterial flora. In addition to members of family Enterobacteriaceae species of Bacillus, Staphylococcus, Streptococcus, and Acinetobacter are the most common in both strains investigated. Gram negative bacteria were increased significantly after feeding on blood meals than those detecting during the feeding on sugar after emergence of the adult female mosquitoes and vise versa with Gram positive bacteria. Bacterial isolates were tested for resistance to the most common commercial antibiotics.     

Production of Antimicrobial Substances by Bacteria Isolated from Fermented Table Olives

Summary The production of antimicrobial substances was studied among 195 bacterial isolates from retail table olives. A total 86 isolates tested positive, and they clustered in 10 groups according to their inhibitory spectra. Many isolates (38.37%) produced strong inhibition against all bacteria tested (Listeria innocua, Lactococcus lactis, Bacillus cereus, B. megaterium, Staphylococcus aureus, Micrococcus luteus, Enterococcus faecalis, and Escherichia coli). The selected bacterial isolates were Gram-positive bacteria with rod morphology (62.67%), short rods (26.65%) or cocci (10.67%). Isolates producing antimicrobial substances may be useful as starters to enhance control of table olive fermentation and improve the safety of retail table olives.     

平湖市第一人民医院儿科患者主要病原菌分布及耐药性分析

目的了解和探讨平湖市第一人民医院儿科感染的常见病原菌的分布状况及耐药情况。方法对该院2012年1月至2012年12月儿科患者送检的合格标本进行细菌培养和药敏试验,并统计分析。结果分离出305株病原菌,其中革兰阳性球菌166株,占54. 5%,革兰阴性杆菌134株,占43. 9%,真菌5株,占1. 6%。革兰阳性球菌中前几位依次为金黄色葡萄球菌82株,占49.4%,人葡萄球菌27株,占16. 3%,表皮葡萄球菌27株,占16. 3%,溶血葡萄球菌17株,占10. 2%。革兰阴性杆菌中前几位依次为肺炎克雷伯菌42株,占30. 2%,大肠埃希菌28株,占20. 1%,鲍曼不动杆菌21株,占15. 1%,阴沟肠杆菌15株,占10. 8%,不同病原菌对常用抗菌药物呈现不同程度的耐药,大肠埃希菌的ESBLs阳性率为28. 6%,肺炎克雷伯菌ESBLs阳性率为11.9%。结论该院儿科患者感染的主要病原菌以革兰阳性球菌为主,最常见为金黄色葡萄球菌,不同病原菌对抗生素的耐药性不同,应加强病原菌的检测及耐药性监测,严格执行消毒灭菌操作的各项规定,尽早进行细菌鉴定和药敏试验,临床应根据药敏结果选择用药,合理使用抗菌药物。     

A note on the isolation and enumeration of Gram positive cocci from marine and estuarine waters

Ninety-two percent of Gram positive cocci detected in estuarine and marine surface waters were identified as Staphylococcus spp. Micrococcus and Streptococcus spp. were rarely isolated from seawater, except at dump site stations located in the Puerto Rico Trench area of the Atlantic Ocean, where they were present in unusually large numbers. Staphylococci were present in greatest abundance near land, but were also found in open ocean surface waters in low, but relatively constant, numbers. Staphylococcus epidermidis , or strains related to Staph. hominis , were present in all samples. Staphylococci from surface water samples collected at the Puerto Rico Trench dump site degraded a wide range of compounds. It is considered that these cocci were derived from the wastes dumped, as very few were found in unpolluted marine waters and in those cases in which stimulatory factors, such as disposal of wastes, do not play a role, Gram positive cocci will not be recovered unless large volumes of water are filtered. The Gram positive cocci are present in very low numbers in natural seawater of the open ocean and, therefore, are commonly overlooked. The significance of these bacteria in the nutrient cycles of the sea remains to be elucidated. Documentation of a widespread occurrence of Gram positive cocci in North Eastern Atlantic seawater is provided.     

烧伤病区感染常见病原菌及抗生素的选择

目的探讨内江市第一人民医院烧伤病区感染的常见病原菌及其对常用抗生素的耐药性．以指导临床抗生素的选择,提出防治措施。方法对1999年1月～2003年12月该院烧伤病区患者因感染而分离出的658株病原菌进行回顾性分析。结果分离出的658株病原菌中．革兰阴性菌425株(64．6％),革兰阳性菌209株(31．8％)．真菌24株(3．6％)。主要病原菌分别为铜绿假单胞菌、金黄色葡萄球菌、大肠埃希菌、凝固酶阴性葡萄球菌和阴沟肠杆菌。药敏提示除真菌外的大多数病原菌出现多重耐药性。革兰阴性菌对亚胺培南、头孢哌酮-舒巴坦、头孢西丁和妥布霉素较为敏感。革兰阳性球菌对替考拉丁和万古霉素高度敏感。结论烧伤感染中以铜绿假单胞菌为主．其次为金黄色葡萄球菌、大肠埃希菌和凝固酶阴性葡萄球菌。了解病原菌的结构及其变化趋势．对于加强抗感染治疗的针对性和主动性,取得最佳的感染防治效果具有重要意义。