MIKRO-LA-TEST系统用于58株葡萄球菌、肠杆菌科细菌鉴定的评价

目的 评价MIKRO-LA-TEST系统对临床常见的葡萄球菌、肠杆菌科细菌的鉴定能力。方法 将新鲜培养的细菌经形态学和／或初步生化鉴定后,制备规定浓度的菌液,接种相应板条,置35～37℃孵育24h。按要求加入生化试剂,判读并记录所有反应结果,经电脑分析后得菌株鉴定结果。结果 58株细菌全部获得属水平的正确鉴定(占100％),除去无法鉴定到种的19株细菌,其余39株中有36株获得种水平的正确鉴定(占92．3％)。结论 与法国生物梅里埃公司VITEK系列、API系统及美国DADE公司MICROSCAN系统相比,MIKRO-LA-TEST系统具有操作简便、成本低的特点,中小型医院可以用其完成临床常见细菌的鉴定。     

MALDI-TOF质谱仪在临床常见细菌鉴定中的应用

目的 探讨两种质谱仪Biotyper MS和Vitek-MS系统在临床常规分离细菌鉴定中的应用价值。方法 收集沈阳军区总医院2012年3月至2013年1月分离自血液、尿液、脑脊液、分泌物和痰液等标本的临床常见菌细菌149株(包括14个菌属和30个菌种)。同时采用两种MALDI-TOF-MS系统对上述菌株进行鉴定,结果与Vitek2compact常规生化鉴定进行比较,对3种方法检测结果不一致菌株用16Sr RNA测序进行确认。结果 在149株常见细菌中,Bruker Biotype属和种的正确鉴定率分别为98%和96%,Vitek-MS属和种的正确鉴定率分别为97%和95%。两者均无错误鉴定菌株。结论 对该组常见细菌,两种质谱仪鉴定结果比较差异无统计学意义(P>0.05),均呈现出优异的鉴定水平,适合用于临床微生物的常规鉴定。     

基质辅助激光解吸电离飞行时间质谱在临床病原菌鉴定中的应用

目的探讨基质辅助激光解吸电离飞行时间质谱(matrix-assisted laser desorption ionization-time of flight mass spectrometry,MALDI-TOF MS)系统用于快速鉴定临床分离菌的可靠性和实用性。方法收集南京军区南京总医院2013年7~10月自临床标本分离的非重复细菌1 061株,分别使用MALDI-TOF MS和Vitek 2 Compact全自动细菌鉴定仪系统进行鉴定,结果不一致的菌株采用16S r DNA测序验证。结果 1 061株临床分离菌中1 058株(99.7%)经MALDI-TOF MS系统正确鉴定到属水平,1 016株(95.8%)正确鉴定到种,其余3株(0.3%)未给出鉴定结果。5株鉴定结果不一致的菌株经16S r DNA测序确认,结果与MALDI-TOF MS和Vitek 2 Compact鉴定符合率分别为40.0%(2/5)和0(0/5)。结论 MALDI-TOF MS可以作为一个快速、准确和价廉的工具应用于临床分离菌的鉴定。     

MALDI-TOF 质谱仪在临床常见细菌鉴定中的应用

目的：探讨两种质谱仪 Biotyper MS 和 Vitek-MS 系统在临床常规分离细菌鉴定中的应用价值。方法收集沈阳军区总医院2012年3月至2013年1月分离自血液、尿液、脑脊液、分泌物和痰液等标本的临床常见菌细菌149株（包括14个菌属和30个菌种）。同时采用两种 MALDI-TOF-MS 系统对上述菌株进行鉴定,结果与 Vitek2 compact 常规生化鉴定进行比较,对3种方法检测结果不一致菌株用16S rRNA 测序进行确认。结果在149株常见细菌中,Bruker Biotype 属和种的正确鉴定率分别为98％和96％,Vitek-MS 属和种的正确鉴定率分别为97％和95％。两者均无错误鉴定菌株。结论对该组常见细菌,两种质谱仪鉴定结果比较差异无统计学意义（P ＞0．05）,均呈现出优异的鉴定水平,适合用于临床微生物的常规鉴定。     

细菌微量快速生化鉴定方法学研究

目的建立细菌微量快速生化鉴定系统,用于临床实验室细菌学的快速诊断。方法应用所研制的27种微量快速生化反应试剂与传统试管方法检测208株临床分离菌及8种质控菌株生化反应符合率,依据杭州24 h的JY Z-15E及JY Z-11E肠杆菌生化编码鉴定管的项目要求组合了该法的肠杆菌15种(RE 15n)及肠杆菌11种(RE 11n)生化鉴定试验条,用208株临床分离菌及8种质控菌菌株进行鉴定试验。结果27种细菌微量快速生化反应试剂与传统试管法生化反应总符合率为93.5%~100%。对208株临床分离菌及8株质控菌的菌种鉴定,其完全符合率15种(RE 15n)为99.9%,11种(RE 11n)为100%。8种质控菌株鉴定完全正确。结论用细菌微量快速生化反应系统鉴定细菌,具有简单、快速的特点,无需特殊设备,成本低廉,所得结果准确、可靠,能够满足临床需要。     

275份胆汁细菌需氧培养及药敏试验

胆道感染是常见急腹症。我们采用胆道手术时无菌抽取胆汁 5～ 8ml直接注入法国生物 -梅里埃公司生产的需氧双相培养瓶 ( HPD) ,对 2 75例急性胆囊结石 ,胆总管结石 ,肝内外胆管结石患者进行胆汁细菌需氧培养 ,并以 API系统鉴定总结如下。一、细菌鉴定结果　 2 75份胆汁分离出病原菌1 65株 (其中 1 0份标本各分离出 2种细菌 ) ,2 5个种 ,阳性率为 56.4%。革兰阴性菌 1 2 5株 ,占 75.8%(其中肠杆菌科细菌 1 0 9株 1 4种 ,占 66.1 % ;非发酵菌 1 6株 3种 ,占 9.7% )。革兰阳性菌 36株 ,占2 1 .8% (其中葡萄球菌 5株 4种 ,占 3.0 % ;肠球菌30…     

自动微生物分析快速链球菌生化鉴定系统鉴定猪链球菌的检测结果评价

目的采用自动微生物分析快速链球菌生化鉴定系统(ATBRapid32Strep)鉴定链球菌,并与国际公认细菌鉴定精标方法API20Strep(链球菌鉴定生化系统)比较,评价检测速度和准确性,使临床微生物室能正确使用ATBRapid32Strep鉴定系统,快速、准确鉴定出各种链球菌,及时为临床提供病原学依据。方法从患者血液、脑脊液,猪心、猪肉等标本中分离出形似链球菌的细菌再次分离分纯,转种于血平板上,置CO2培养箱,35℃24h培养后,用ATBRapid32Strep和API20Strep、传统手工方法分别对18株细菌进行鉴定和药敏试验。结果ATBRapid32Strep鉴定出16株猪链球菌Ⅱ型,2株牛链球菌Ⅱ型;API20Strep鉴定出17株猪链球菌Ⅱ型,1株不能鉴定;传统手工鉴定18株均为D群非肠球菌。这18株菌对氯霉素、克林霉素、红霉素、左氧氟沙星、青霉素、奎奴普叮/达福普叮、头孢噻肟、复方新诺明、万古霉素均敏感,对四环素耐药。结论ATBRapid32Strep4h能鉴定出猪链球菌,与API20Strep鉴定符合率为94.4%,是一种操作简单、快速、准确的细菌鉴定系统,能快速为临床提供感染性疾病病原学信息,满足临床医生和微生物检验人员工作需要。     

用gyrB、rpoD基因扩增测序进行气单胞菌分类的探讨

目的获得气单胞菌更准确的分类结果。方法收集经Vitek全自动细菌分析仪及配套的GN鉴定卡鉴定为气单胞菌的菌株231株,对其gyr B、rpo D两个管家基因进行PCR扩增及测序,并与Gen Bank中标准序列进行基因序列比对和单核苷酸替代率分析,综合分析gyr B、rpo D两基因测序结果并进行基因分类。结果 231株气单胞菌中,69株(29.9%)gyr B和rpo D基因均获得测序结果,86株(37.2%)仅获得gyr B序列,50株(21.6%)仅获得rpo D序列,26株(11.3%)两种基因均未获得序列。生化表型鉴定为嗜水气单胞菌的菌株中62.7%基因分类结果为达卡气单胞菌,仅22.4%为嗜水气单胞菌,其种间生化反应结果相近。结论气单胞菌属的生化分类与基因分类结果存在差异。达卡气单胞菌与嗜水气单胞菌的鉴定依赖于基因序列分析。     

一株临床不常见念珠菌的检验

目的对1株临床不常见念珠菌的检测进行探讨。方法采用4种不同方法(ATB、DADE、Remel和分子生物学方法)对1株不常见念珠菌进行检测。结果ATB、Remel系统检测结果为邹褶念珠菌;DADE系统检测结果为克柔念珠菌;分子生物学检测结果为近邹褶念珠菌。讨论对于临床不常见的念珠菌,仅用生化反应鉴定是不够的,为了正确鉴定到种,必要时还要用分子生物学方法作核酸序列分析。     

2199株临床常见病原菌动态分析

目的探讨临床常见病原菌及其对抗生素的耐药性,分析临床致病菌的分布状况和对各种抗生素的耐药趋势。方法采用美国德灵公司自动细菌鉴定及药敏分析仪进行鉴定和药敏试验。结果2199株病原菌中,细菌占83%,真菌占17%,以大肠埃希氏菌最为常见。结论随着临床抗生素的泛用和滥用病原菌的分布与构成不断地发生变化,病原微生物的耐药状况十分严重。     

2011年临床病原菌分布及耐药性分析

目的 分析某院2011年上半年临床分离病原菌的分布及耐药性.方法 采用MicroSCcan-WalkAway 40SI型自动鉴定及药敏测试系统进行菌种鉴定及药敏试验.结果 检出率前5位病原菌分别为大肠埃希菌26.07%(91株)、金黄色葡萄球菌23.78%(83株)、表皮葡萄球菌20.06%(70株)、铜绿假单胞菌16.61%(58株)、鲍曼不动杆菌13.47%(47株);病原菌敏感率大于或等于75%的抗菌药物包括亚胺培南(大肠埃希菌、铜绿假单胞菌、鲍曼不动杆菌),哌拉西林/他唑巴坦(铜绿假单胞菌、金黄色葡萄球菌),复方新诺明(嗜麦芽窄食单胞菌),丁胺卡那霉素(鲍曼不动杆菌),利福平、奎奴普丁/达福普丁、吗啉恶酮、万古霉素(金黄色葡萄球菌).结论 该院常见病原菌的耐药性持续增强,应科学规范使用抗菌药物.     

耐亚胺培南鲍曼不动杆菌耐药表型和碳青霉烯酶基因型分析

目的检测广州市第一人民医院20株耐亚胺培南鲍曼不动杆菌的耐药性,研究并分型其碳青霉烯酶基因。方法用法国生物梅里埃公司VITEK2全自动细菌鉴定仪进行菌株鉴定和药敏试验,用6种碳青霉烯酶特异性基因引物进行PCR扩增和基因型的测序分析,并通过网上GenBank进行比对以确定编码酶基因的类型。结果 20株鲍曼不动杆菌对哌拉西林-他唑巴坦、左氧氟沙星和阿米卡星的耐药率分别为85%、50%和25%。对其他所测抗菌药的耐药率均在90%以上。扩增结果显示17株(85%)细菌携带碳青霉烯酶OXA-23基因,16株(80%)携带OXA-51基因,1株(5%)携带OXA 58基因。OXA-24、VIM、IMP基因引物PCR扩增均为阴性,随机抽取OXA-23基因阳性株进行双向测序后在网上GenBank比对与OXA-23标准株99%同源,OXA-51基因阳性株与OXA-66标准株99%同源,OXA-58基因阳性株与OXA-58标准株99%同源。结论我院耐碳青霉烯类抗生素的鲍曼不动杆菌呈现出多重耐药现象,对阿米卡星的耐药率最低,OXA-23型碳青霉烯酶基因检出率最高,OXA-23/OXA-51类基因占60%,应引起临床高度重视,防止在医院内广泛传播。     

Identification of oral bacteria by 16S rRNA gene analysis in elderly persons requiring nursing care

After incubation of saliva from 58 semi-bedridden elderly persons, the cultures were identified based on the 16S rRNA gene base sequence to compare the identification by the conventional culture method. As a result, the 16S rRNA gene base sequence of 198 strains identified by the culture method showed 98.5% or more homology in some of the Human Oral Microbiome database, and the identification of bacterial species and genus was possible. When an organism identified by the 16S rRNA gene sequencing method was compared with that by the culture method, the concordance rates were 54.5% at the genus level and 35.9% at the species level. Streptococcus mitis strains most frequently isolated from saliva that were identified by the culture method were identified as the same species by the 16S rRNA gene sequencing method (32/35), and all the 11 Streptococcus salivarius strains identified by the culture method were identified as the same species by the 16S rRNA gene sequencing method. All the strains identified as Streptococcus anginosus group by the culture method and 8 of the 9 strains identified as Prevotella species by the culture method were identified as the same group and genus by the 16S rRNA gene sequencing method. When an oral microbial flora test with saliva samples from elderly persons is performed, the 16S rRNA gene sequence identification enables us to identify major indigenous bacteria and pathogenic bacteria and is considered useful as a means of supplementing the conventional culture method.     

Secondary and tertiary prevention in the management of low-trauma fracture

A significant risk factor for osteoporotic fracture is a previous atraumatic fracture. The objective of this study was to investigate whether patients with Colles fracture from minimal trauma were subsequently identified, assessed and treated for their elevated risk of fracture. Medical records at Sir Charles Gairdner Hospital in Perth, Western Australia, from August 1999 to July 2000 were audited and 111 patients who had sustained a Colles fracture from minimal trauma were identified. Questionnaires were subsequently posted to participants to determine whether any assessment or treatment was undertaken outside the hospital system. According to documentation in the medical records, 9% (10/111) had their bone mineral density assessed, 15% (17/111) were receiving medical therapy for osteoporosis, 7% (8/111) had their falls risk assessed and 51% (58/111) were seen by a physiotherapist. Of the 58 who received physiotherapy, 76% (44/58) received upper limb exercises and 19% (11/58) received lower limb or balance exercises. Follow-up questionnaires one to two years after the fracture were returned by 43% (48/111) of the sample. By this time, 37% (18/48) had BMD assessed and 27% (13/48) were receiving medical therapy for osteoporosis. Thirty-five per cent (17/48) of patients recalled being advised to increase their calcium intake. Of those who reported more than one fall during the past 12 months, 62% (8/13) had been seen by a physiotherapist, 46% (6/13) reported having their balance assessed and 54% (7/13) reported having a home visit for assessment of rails etc. Despite the availability of effective treatments, a substantial proportion of patients with Colles fracture from minimal trauma are not all being identified, assessed or treated for their elevated risk of subsequent osteoporotic fracture.     

Comparative in vitro activities of new fluorinated quinolones and other antibiotics against coagulase-negative Staphylococcus blood isolates from neutropenic patients, and relationship between susceptibility and slime production.

We evaluated the in vitro antibiotic susceptibilities of 31 coagulase-negative Staphylococcus isolates causing septicemia in neutropenic patients undergoing norfloxacin prophylaxis. All the strains but one were resistant to 1 microgram of norfloxacin per ml. At the same concentration, ciprofloxacin, ofloxacin, imipenem, and pefloxacin were inhibitory for 19 (61%), 19 (61%), 18 (58%), and 14 (45%) of the evaluated strains, respectively. Imipenem had an MBC/MIC ratio of greater than or equal to 32 against 19 (61%) of the evaluated isolates, and resistant subpopulations were detected at 5 micrograms/ml in 16 of 17 oxacillin-resistant strains and in 3 of 14 oxacillin-susceptible or -tolerant strains. Resistance to gentamicin was seen with increased frequency among slime-producing strains.     

Trimethoprim resistance amongst urinary pathogens in south India

Two hundred and eighty four strains of Enterobacteriaceae, responsible for significant bacteriuria, were isolated, over a three month period, in Vellore, India. Sixty-four per cent of these strains were resistant to 10 mg/l of trimethoprim. Moreover, this population was dominated by high level resistance (minimum inhibitory concentration greater than 1000 mg/l) and these accounted for 57.3% of all strains studied. Over half of the resistant strains were able to transfer trimethoprim resistance to standard Escherichia coli strains. However, the high incidence of transferable resistance did not result from the spread of one plasmid type as 58 different plasmid types were identified. These results are in marked contrast to recent findings in Europe where the incidence of high level transferable trimethoprim resistance is falling.     

脑梗死患者并发尿路感染的病原菌分布及耐药性分析

目的:探讨脑梗死患者并发尿路感染的病原菌分布及耐药性.方法:回顾性分析2020年6月～2020年9月就诊于我院的脑梗死并发尿路感染住院患者的尿培养细菌学资料.结果:纳入研究207例患者中共分离出非重复病原菌214株,以革兰阴性菌为主,占66.4％(139/214),主要菌种依次为:大肠埃希菌、肺炎克雷伯菌、铜绿假单胞菌...     

MALDI-TOF MS直接鉴定血培养阳性病原的临床应用

目的了解基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)直接快速检测血培养报阳样本的可行性。方法收集同济大学附属同济医院报阳血培养样本682例,从血培养瓶直接抽取血液至分离胶采血管离心富集细菌后,采用MALDI-TOF MS进行直接鉴定(快速质谱法)。同时对报阳血培养标本进行涂片镜检及转种培养后,分别采用Vitek 2 Compact自动化鉴定药敏仪及MALDI-TOF MS对培养菌落进行鉴定,比较两者鉴定结果的一致性,结果不一致的菌株采用16S rDNA基因测序进行验证。结果682例血培养阳性样本中,有664例检出单数菌,其中539例(81.2%)快速质谱法可准确鉴定(种/属),122例(18.4%)未鉴定出(种/属),仅3例(0.5%)鉴定错误,分别为泛菌属、肺炎克雷伯菌和头状葡萄球菌。664株菌株中,革兰阴性菌367株,革兰阳性菌286株,念珠菌11株,以转种培养后质谱鉴定结果为标准,革兰阴性菌、革兰阳性菌、念珠菌快速质谱法的鉴定准确率(种/属)分别为85.8%/6.0%、65.0%/3.1%、63.6%/0;革兰阴性杆菌的鉴定准确率显著高于革兰阳性球菌和真菌,差异有统计学意义(χ²=57.967,P<0.01;χ²=7.21,P<0.05)。革兰阴性菌中,肠杆菌科、非发酵菌及其他革兰阴性菌鉴定准确率(种/属)分别为89.0%/5.0%、65.0%/10.0%、66.7%/22.2%。革兰阳性菌中,葡萄球菌属、肠球菌属、链球菌属和其他革兰阳性菌鉴定准确率(属/种)分别为70.9%/1.1%、71.4%/2.4%、40.5%/10.8%、50%/7.1%。有18例样本检出复数菌,快速质谱法从其中14例中检出复数菌,鉴定准确率为77.8%,均只鉴定到种。结论快速质谱法检测血培养报阳样本具有可行性,对单数菌的鉴定错误概率非常低,且对临床常见病原菌鉴定准确率均>70%。但对复数菌鉴定效果不佳。临床可根据实际需求选择合适的细菌鉴定方法。     

肺炎患者痰液分离鲍曼不动杆菌耐药基因分析

目的:分析呼吸内科肺炎患者痰液分离鲍曼不动杆菌携带β-内酰胺酶基因及主动外排泵基因情况。方法:收集经生化鉴定为醋酸钙-鲍曼不动杆菌复合体的菌株160株,将鲍曼不动杆菌特异性片段(Ab-ITS)的二重PCR法确认为鲍曼不动杆菌者纳入研究。用PCR法检测鲍曼不动杆菌β-内酰胺酶基因和主动外排泵基因,包括A类β-内酰胺酶(blaTEM、blaPER和blaCARB)、B类β-内酰胺酶(blaIMP和blaVIM)、C类β-内酰胺酶(blaADC和blaDHA)、D类β-内酰胺酶(blaOXA-23、blaOXA-24、blaOXA-51和blaOXA-58)和主动外排泵相关基因(adeB、adeJ、macB、emrB、emrA、abeS、abeM和craA)。以微量肉汤稀释法检测加入外排泵抑制剂羰基氰化氯苯腙(CCCP)前后,携带adeB基因鲍曼不动杆菌对亚胺培南的最低抑菌浓度(MIC)的变化。结果:临床分离160株醋酸钙-鲍曼不动杆菌复合体中鲍曼不动杆菌143株(占90%),包括亚胺培南耐药株119株(占83.2%)和敏感株24株(占16.8%)。所有鲍曼不动杆菌均检出blaOXA-51基因,其中,亚胺培南耐药株中检出blaOXA-23(98.3%)、blaTEM(41.4%)、blaADC(98.3%)和blaDHA(0.7%),blaPER、blaCARB、blaIMP、blaVIM、blaOXA-24和blaOXA-58均未检出;敏感株中未检出blaOXA-23。adeJ、macB、abeM和craA基因在143株鲍曼不动杆菌中的携带率均为100%;emrB、emrA和abeS基因携带率均超过80%。adeB基因在亚胺培南耐药株中检出117株(98.3%)。在CCCP干预试验中,携带adeB基因的鲍曼不动杆菌MIC50降低1/2。结论:呼吸内科鲍曼不动杆菌主要携带blaOXA-23和adeB主动外排泵基因,可能与亚胺培南耐药密切相关。     

72株沙门菌血清型分布及药敏试验结果分析

目的：了解广东东莞地区沙门菌血清型分布,腹泻患者沙门菌检出数量及药敏试验结果,为该省今后的疾病防控,药物治疗提供参考依据。方法收集东莞市人民医院2013年4—12月门诊及住院患者腹泻粪便标本1150份,用改良亚硒酸盐磺绿增菌肉汤和沙门菌显色平皿分离检测沙门菌;药敏试验采用法国生物梅里埃ATB微生物鉴定和药敏分析系统及其配套药敏试条。结果共检出沙门菌72株,检出率为6．3％,分出21个血清型,以鼠伤寒沙门菌检出数量最多,22株（30．5％）,肠炎沙门菌16株（22．2％）。年龄分布1周岁以下42株（58．3％）,3-10岁12株（16．7％）,20岁以上18株（25．0％）。药敏试验结果耐药率较高的药物有哌拉西林（50．0％）,替卡西林（48．6％）。结论本地区腹泻患者检出沙门菌属以鼠伤寒沙门菌检出率最高,肠炎沙门菌次之,年龄1周岁以下较多。临床医师应加强对腹泻患者沙门菌属监测,尤以婴幼儿为重点,早期发现早期治疗以减少并发症发生。