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     

厌氧菌微量生化板的研制

利用预成酶的原理,研制厌氧菌微量化生鉴定板,用国际标准菌株检测其符合率达９５．９％,重复率达９６．３３％,用己知引进参考菌株检测,其符合率达９４．８４％,重复率达９４．３３％,而对临床分离的革兰阴性细菌的符合率达８１％,对临床分离的革兰阳性细菌符合率达９０％,此法效果与国外ＡＰＩ－２０Ａ产品效果类似,但此法具有快速,操作简便,重复性好等优点,值得在广大临床检测中推广和普及。     

VITEK-2 compact全自动微生物鉴定仪对葡萄球菌鉴定能力的评价

目的评价VITEK-2 compact全自动微生物鉴定仪对葡萄球菌的鉴定能力。方法收集从我院病人标本中分离的葡萄球菌81株。常规细菌培养后,用VITEK-2 compact和API Staph系统进行检测,以API Staph系统为参照,评价VITEK-2 compact的优势和不足。结果 VITEK-2 compact和API Staph系统的总体鉴定符合率为95.1%,其中金黄色葡萄球菌的鉴定符合率为100%,凝固酶阴性葡萄球菌的鉴定符合率为90.5%。结论 VITEK-2 compact鉴定系统能够满足临床工作的需求,其中对金黄色葡萄球菌的鉴定率较高,在进行凝固酶阴性葡萄球菌鉴定时有一定的局限性,需要其他方法予以补充。     

API20E鉴定系统在肠杆菌科细菌检验中的应用

随着致人类感染的致病微生物种、属的不断增加,国内外新的生化鉴定系统不断推出,以传统试验来鉴定微生物已不能适应鉴定的需要,而成套鉴定系统及编码鉴定方法,使细菌鉴定简易化。微量化和快速化。在目前众多的细菌鉴定系统中,API细菌鉴定系统在世界范围内应用较广,已被国际微生物学家所公认。近2年来我们应用Anl鉴定技术鉴定了48株肠杆菌科细菌,与国产细菌微量生化鉴定管GYZ－15e做了比较,取得了满意的效果。1材料与方法三．且材料API20E生化反应试条（法国海里埃公司生产）;API20EPI。US软件（法国梅里埃公司生产）;细菌…     

基于数值鉴定法的肠杆菌科生化试剂盒的研制及效果评价

【背景】肠杆菌科细菌是食源性和临床感染的主要病原菌,对人类和动物健康有重要影响,以API 20E为“金标准”的数值鉴定法是其主要的鉴定方法之一,但现有的数值鉴定方法存在手工单个加样或/和准确率低、价格贵的问题。【目的】研制出半自动化、准确性高及价格低的肠杆菌科细菌数值鉴定生化试剂盒。【方法】在本团队前期建立的肠杆菌科数值鉴定系统理论模型与配套软件的基础上,设计并优化筛选出的24种生化基质微量化配方,在此基础上,研制半自动冻干鉴定条;以现行商品化的数值鉴定条API 20E、MALDI-TOF MS及16S rRNA基因测序为对照,对研制的集半自动生化鉴定条与在线分析软件于一体的生化试剂盒进行效果评价。【结果】共获得458株肠杆菌科细菌的生化谱,在属水平总体鉴定准确率达到98.5%,在种水平总体鉴定准确率达到96.5%;仅需两次加样即可获得鉴定结果,且价格仅为API 20E的4.46%;产品保质期为7 d,生化试验具有稳定可重复性。【结论】本研究研制出的半自动化肠杆菌科细菌数值鉴定生化试剂盒简便经济、准确率高,为肠杆菌科细菌的鉴定提供技术支撑,并为其他科属细菌的数值鉴定生化产品的开发提供技术指引。     

四株鸡有益链球菌的分离鉴定

用厌氧菌增菌培养基（ＢＲ）、厌氧菌血琼脂（ＣＤＣ）、选择乳酸杆菌培养基（ＬＢＳ）从鸡直肠内容物中分离出四株革兰氏阳性链球菌,据分离菌的形态、染色和培养特性及生化试验结果,鉴定为牛粪链球菌两株（Ｓｂ１、Ｓｂ２）,尿链球菌两株（Ｓｆ１、Ｓｆ２）。四株菌对雏鸡安全,无致病性和副作用,对头孢三嗪、复方新诺明耐药。     

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。     

梅里埃VITEK 2全自动微生物鉴定药敏分析系统性能评估

评价梅里埃VITEK 2全自动微生物鉴定药敏分析系统的性能。选取卫生部室间质评菌株20株，统计分析鉴定结果的正确性、药敏结果。结果：鉴定重复率为100%，正确率为100%，药敏结果符合率为100%。梅里埃VITEK 2比VITEK 2 COMPACT全自动微生物鉴定及药敏分析系统操作更便捷、更准确率，微生物检测质量更佳。     

阪崎克罗诺杆菌的ITS序列分析

摘 要：以分离自不同来源（不同地区乳制品及某一企业婴儿配方乳粉加工生产环境）、经API20E鉴定为阪崎克罗诺杆菌的37株分离株为研究对象,采用ITS序列对其进行鉴定分析。结果表明,ITS序列分析在阪崎克罗诺杆菌鉴定中具有一定的优越性,且研究发现,ITS序列中含有两种功能基因tRNAgul和tRNAile+ala,这为今后阪崎克罗诺杆菌的鉴定及ITS基因的进一步分析提供了可供参考的理论依据。     

国内首例皮瘤丝孢酵母真菌血症分离株的表型及rDNA序列鉴定

目的通过对1例“脂膜炎”患者外周血分离株的表型及分子生物学鉴定,报道国内首例皮瘤丝孢酵母菌血症。方法采集患者的血及鼻咽结节组织标本通过真菌培养获得分离株,经沙氏培养基、马铃薯培养基、科玛嘉念珠菌显色培养后形态学观察及API 20C AUX系统鉴定,最后经PCR扩增rDNA基因测序证实。采用CLSI制定的M27-A2微量稀释法对病原菌株进行7种药物的体外药敏试验。结果分离株通过表型和分子生物学鉴定为“皮瘤丝孢酵母”。药敏结果显示菌株对两性霉素B耐药,氟康唑、伏立康唑、伊曲康唑、5-氟胞嘧啶敏感。卡泊芬净的抑菌浓度较低,而米卡芬净的抑菌浓度相对较高。患者因多种并发症死亡。结论皮瘤丝孢酵母引起的真菌血症为我国首例报道。API 20C AUX酵母鉴定系统可作为鉴定皮瘤丝孢酵母快速而简便的方法。通过分子生物学DNA序列检测分析能鉴定皮瘤丝孢酵母。     

Comparison of different biochemical and molecular methods for the identification of Vibrio parahaemolyticus

AIMS: Multicentre evaluation of biochemical and molecular methods for the identification of Vibrio parahaemolyticus. METHODS AND RESULTS: For the biochemical identification methods, API 20E and API 20NE and Alsina's scheme were evaluated in intra- and interlaboratory tests in order to determine the accuracy and concordance of each method. Both in intra- and interlaboratory tests, the Alsina's scheme showed the highest sensitivity (86% of correct identifications in the interlaboratory test). False-positive results were obtained by all methods (specificity was 95% for API 20E, 73% for API 20NE and 84% for Alsina's scheme) and concordance varied from 65% of API 20NE to 84% of API 20E. For the molecular identifications, polymerase chain reaction (PCR) for the detection of toxR gene, tl gene and pR72H fragment were tested on 30 strains by two laboratories. The PCR for toxR showed the highest inclusivity (96%), exclusivity (100%) and concordance (97%). CONCLUSIONS: Among the biochemical identification methods tested, the Alsina's scheme gave more reliable results; however, in order to avoid false-positive results, all the biochemical identifications should be confirmed by means of molecular methods. SIGNIFICANCE AND IMPACT OF THE STUDY: Availability of an efficient identification method of Vibrio parahaemolyticus to use in official control of fisheries products.     

Biochemical properties of Corynebacterium amycolatum strains

C. amycolatum is the most commonly isolated nonlipophilic species of Corynebacterium from clinical samples. However, the lack of good commercial identification tests in microbiology laboratories causes some difficulties in C. amycolatum diagnostics. We decided to examine biochemical and enzymatic properties of isolated strains and analyze the occurrence of particular biochemical profiles (biotypes). Perhaps it would let improve the identification schemes. 70 strains of C. amycolatum were analyzed. The estimation of biochemical properties consisted of the results of API Coryne and API ZYM tests (bioMérieux), the ability of excreting of protease, esterase, lipase and lecithinase. Analyzed strains had various biochemical and enzymatic properties. Almost all strains fermented glucose (98.6%) and maltose (95.7%) and produced pyrasinamidase (94.3%). All strains produced alkaline phosphatase and phosphohydrolase, and 95.7%--acid phosphatase. Biotypes of particular strains were determined on the biochemical reactions included in the API Coryne tests. In the group of 70 strains 21 profiles were distinguished among which 3100325 biotype (35.7%) was dominant. The lipolysis was defined on Tween 20, Tween 40, Tween 60, Tween 80 medium and with the API ZYM test usage. All strains produced esterase-lipase (esterase C-8), 95.7% of strains-esterase C-4, and 21.4% lipase C-14. Among analyzed strains 18.6% hydrolyzed Tween 20, 14.3% Tween 60, and 1.4% Tween 40. None of these strains demonstrated lipase and lecithinase activity. Difficulties in concerning C. amycolatum as pathogens justify further investigations.     

A comparison of three identification kits for the confirmation of Aeromonas spp.

A comparative study was made of the ability of three commercial identification kits to confirm the identity of motile aeromonads isolated from foods. The kits included the API 20E, API 20NE and Microbact 24E. The results showed that both the API 20NE and Microbact 24E correctly identified 97.5% of isolates but the API 20E only 72.5%.     

Comparison of biochemical and molecular methods for the identification of bacterial isolates associated with failed loggerhead sea turtle eggs

Aims: Comparison of biochemical vs molecular methods for identification of microbial populations associated with failed loggerhead turtle eggs. Methods and Results: Two biochemical (API and Microgen) and one molecular methods (16s rRNA analysis) were compared in the areas of cost, identification, corroboration of data with other methods, ease of use, resources and software. The molecular method was costly and identified only 66% of the isolates tested compared with 74% for API. A 74% discrepancy in identifications occurred between API and 16s rRNA analysis. The two biochemical methods were comparable in cost, but Microgen was easier to use and yielded the lowest discrepancy among identifications (29%) when compared with both API 20 enteric (API 20E) and API 20 nonenteric (API 20NE) combined. A comparison of API 20E and API 20NE indicated an 83% discrepancy between the two methods. Conclusions: The Microgen identification system appears to be better suited than API or 16s rRNA analysis for identification of environmental isolates associated with failed loggerhead eggs. Significance and Impact of the Study: Most identification methods are not intended for use with environmental isolates. A comparison of identification systems would provide better options for identifying environmental bacteria for ecological studies.     

The isolation and characterization of the aerobic endospore-forming bacteria present in the liquid phase of an anaerobic fixed-bed digester,while treating a petrochemical effluent

Sixty-nine gram-positive endospore-forming rods were isolated from the liquid phase of an anaerobic digester, while treating a fatty acid-rich petrochemical effluent. These strains, including eight reference strains, were characterized and the similarities between the different strains were calculated using Sokal and Michener's simple matching coefficient. Phenotypic characteristics, determined by the API 20E and API 50CHB galleries, other biochemical tests, and morphological characteristics, were used for the numerical analysis. The strains were grouped into 12 (five major and seven minor) clusters. Nine of the clusters were positively identified asBacillus pumilus, B. subtilis, B. sphaericus, B. laterosporus, B. brevis, B. cereus, B. coagulans, B. megaterium, andB. circulans. Three clusters could not be identified using Gordon's classical system or the API identification system. Most of the aerobic endospore-forming rods (72%) utilized both acetic and propionic acid, and 17% utilized acetic acid as carbon source, but only under aerobic conditions. A small percentage of the strains studied (11%) was unable to utilize the fatty acids present in the petrochemical substrate, and no explanation could be given as to how they obtained their carbon source. Seventy-eight percent of the strains did not show growth in anaerobic agar. It was possible that sufficient oxygen, required for growth by these members of the genusBacillus, was introduced by the substrate. Since ample time had been allowed for population selection, their presence indicates that these aerobic strains can survive, grow, and compete in the digester environment but their relative importance and role in the primary digestion reactions is not clear.     

Differences in the API 20E biochemical patterns of clinical and environmental Vibrio parahaemolyticus isolates

Genetic differences in clinical and environmental strains of Vibrio parahaemolyticus have been widely used as criteria in identifying pathogenic isolates. However, few studies have been carried out to assess the differences in biochemical characteristics of V. parahaemolyticus isolates from human and environmental sources. We compared the biochemical profiles obtained by the characterization of V. parahaemolyticus isolates from human infections and the marine environment using the API 20E system. Environmental and clinical isolates showed significant differences in the gelatin and arabinose tests. Additionally, clinical isolates were correctly identified according to the API 20E profile using 0.85% NaCl diluent, but they presented nonspecific profiles with 2% NaCl diluent. In contrast, use of 2% NaCl diluent facilitated correct identification of the environmental isolates. Clinical isolates showed significant differences in up to five biochemical tests with respect to the API 20E database. The API 20E system is widely used in routine identification of bacteria in clinical laboratories, and this discrepancy in an important number of biochemical tests may lead to misidentification of V. parahaemolyticus infection.     

Identification of Clostridium botulinum with API 20 A, Rapid ID 32 A and RapID ANA II

Three commercially available test systems for the identification of anaerobic bacteria were evaluated for the identification of 18 proteolytic group I and 69 non-proteolytic group II Clostridium botulinum, four Clostridium sporogenes and 18 non-toxigenic group II C. botulinum-like strains. All proteolytic C. botulinum strains were misidentified by the Rapid ID 32 A and RapID ANA II, while 14 strains and all C. sporogenes strains were identified as C. botulinum or C. sporogenes by the API 20 A. Reversely, all non-proteolytic C. botulinum strains were misidentified by the API 20 A while the Rapid ID 32 A recognized 67 and RapID ANA II 68 strains. All C. sporogenes strains were recognized by the RapID ANA II, while the Rapid ID 32 A recognized one strain. All non-proteolytic non-toxigenic strains were identified as C. botulinum group II by the Rapid ID 32 A, 17 strains by the RapID ANA II, and one strain by the API 20 A. The results show that these test systems do not provide a reliable method for identification of C. botulinum.     

Comparison of rapid NFT and API 20E with conventional methods for identification of gram-negative nonfermentative bacilli from pharmaceuticals and cosmetics

The accuracy of the Rapid NFT and the API 20E identification systems was evaluated by comparing them with conventional biochemical methods for the identification of gram-negative, nonfermentative bacilli. The organisms were recovered from preserved, nonsterile pharmaceutical and cosmetic products. A total of 123 test isolates that are commonly encountered in these products were used. By using the criteria of accurate and reliable identification without employing additional tests, Rapid NFT was found to be more accurate after 48 h of incubation than API 20E for characterizing isolates to the species level. Therefore, close agreement between NFT and conventional methods for identification of industrial gram-negative isolates provides evidence that the Rapid NFT system is an improved and rapid method for identifying these organisms to the species level with minimal use of supplementary tests.     

Comparison of rapid NFT and API 20E with conventional methods for identification of gram-negative nonfermentative bacilli from pharmaceuticals and cosmetics.

The accuracy of the Rapid NFT and the API 20E identification systems was evaluated by comparing them with conventional biochemical methods for the identification of gram-negative, nonfermentative bacilli. The organisms were recovered from preserved, nonsterile pharmaceutical and cosmetic products. A total of 123 test isolates that are commonly encountered in these products were used. By using the criteria of accurate and reliable identification without employing additional tests, Rapid NFT was found to be more accurate after 48 h of incubation than API 20E for characterizing isolates to the species level. Therefore, close agreement between NFT and conventional methods for identification of industrial gram-negative isolates provides evidence that the Rapid NFT system is an improved and rapid method for identifying these organisms to the species level with minimal use of supplementary tests.