无菌体液与非无菌体液细菌耐药监测对比分析

目的探讨该院2014年无菌体液与非无菌体液来源细菌的菌种分布及耐药性的差异。方法采用回顾性分析方法,应用BD Phoenix~(TM)100全自动细菌鉴定药敏系统进行细菌鉴定与药敏检测,应用Whonet5.6软件和SPSS19.0统计软件对细菌耐药率进行统计分析。结果无菌体液分离率最高的细菌为大肠埃希菌(43%),非无菌体液分离率最高的细菌为铜绿假单胞菌(21%)。无菌体液来源的大肠埃希菌对氨苄西林、氨曲南、环丙沙星、氯霉素等11种抗菌药物耐药率低于非无菌体液来源的菌株,无菌体液来源的金黄色葡萄球菌对阿米卡星、阿莫西林/克拉维酸、环丙沙星等6种抗菌药物耐药率低于非无菌体液来源的菌株,无菌体液来源的铜绿假单胞菌对氨曲南耐药率低于非无菌体液来源的菌株,无菌体液来源的肺炎克雷伯菌对氨苄西林/舒巴坦、复方新诺明、氯霉素等6种抗菌药物耐药率低于非无菌体液来源的菌株,差异均有统计学意义(P0.05)。结论无菌体液与非无菌体液来源细菌的菌种分布及耐药性存在差异,加强无菌体液细菌耐药性监测尤为重要。     

The performance of 4 different supplements and 5 blood culture bottles types in detection of bacteria and Candida spp. in simulated sterile body fluid cultures

The purpose of this investigation was to evaluate the performance of 4 supplements: horse blood, fastidious organisms supplement (FOS), haemin isovitalex albumine (HIA), and brain heart infusion–haemin isovitalex albumine (BHI-HIA) and 5 blood culture bottles: Bactec Mycosis IC/F, Plus Aerobic/F, Peds Plus/F from the Bactec 9240 system, and BacT/Alert FA and BacT/Alert PF from the BacT/Alert 3D system, in detection of bacteria and Candida spp. in simulated sterile body fluids other than blood models. In total, 8 reference strains (Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus pneumoniae, Neisseria meningitidis, Haemophilus influenzae, Listeria monocytogenes, Candida albicans, and Candida parapsilosis) and 11 clinical bacteria and yeast isolates (6 isolates from cerebrospinal fluid and 5 isolates from blood) were included in this study. Horse blood, FOS, and HIA were significantly better than no supplements (P < 0.0001, P < 0.0002, and P = 0.05, respectively) in detection of bacteria. Interestingly, there was no significant difference between BHI-HIA and bottles without any supplements. Sixty bottles analyzed of which 59 (98.33%) bottles with horse blood, 53 (88.33%) with FOS, 45 (75.00%) with HIA, and 43 (71.67%) with BHI-HIA signaled positive. The positivity rates with horse blood were significantly higher than with HIA and BHI-HIA (P < 0.0005 and P < 0.0001, respectively). Similarly, the blood culture bottles with horse blood had shorter time to detection (TTD) compared to bottles with FOS and HIA (P < 0.05 and P < 0.0001, respectively). When yeasts were analyzed, almost all (124/125) blood culture bottles with Candida spp. signaled positive even in the absence of supplements. Bactec Mycosis IC/F had significantly shorter TTD compared to Bactec Peds Plus/F, Bactec Plus Aerobic/F, BacT/Alert FA, and BacT/Alert PF bottles in detection of Candida spp. (P < 0.005, P < 0.05, P < 0.001, and P < 0.001, respectively). The present study showed that horse blood was the most effective supplement in growth of bacteria in the blood culture bottles that were analyzed in the study.     

2010年中国CHINET临床分离自无菌体液的细菌分布和耐药性监测

目的了解国内不同地区14所教学医院2010年从临床分离自脑脊液及其他无菌体液(胸水、腹水、胆汁等)的细菌分布和对抗菌药物的耐药性。方法共收集2010年临床分离的无菌体液2409株非重复的细菌,采用纸片扩散法进行药敏试验,结果按CLSI 2010年版标准判读药敏结果,采用WHONET 5.4软件进行数据分析。结果 2409株非重复的细菌中,革兰阴性菌1 353株,占56.2%,革兰阳性菌1 056株,占43.8%。脑脊液中最常见的细菌分别为:凝固酶阴性葡萄球菌(CNS)、鲍曼不动杆菌、肺炎克雷伯菌、金葡菌和大肠埃希菌。其他无菌体液中最常见的细菌分别为:大肠埃希菌、CNS、屎肠球菌、肺炎克雷伯菌和铜绿假单胞菌。脑脊液标本中MRSA和MRCNS的检出率分别为78.6%和69.7%。其他无菌体液标本中MRSA和MRCNS的检出率分别为60.3%和67.2%。未发现对万古霉素、替考拉宁和利奈唑胺耐药的葡萄球菌。脑脊液标本中未发现对万古霉素耐药的肠球菌属细菌,在其他无菌体液中发现17株对万古霉素耐药的屎肠球菌。脑脊液标本中产ESBLs大肠埃希菌和克雷伯菌属细菌(肺炎克雷伯菌和产酸克雷伯菌)的检出率分别为62.5%和30.0%,其他无菌体液标本中产ESBLs大肠埃希菌和克雷伯菌属细菌(肺炎克雷伯菌和产酸克雷伯菌)的检出率分别为50.4%和27.3%。脑脊液标本中泛耐药鲍曼不动杆菌、肺炎克雷伯菌和铜绿假单胞菌分别检出10株(18.2%)、8株(27.6%)和1株(7.7%)。其他无菌体液标本中泛耐药鲍曼不动杆菌、肺炎克雷伯菌、铜绿假单胞菌和大肠埃希菌分别检出43株(36.8%)、18株(11.5%)、3株(2.1%)和1株(0.2%)。结论脑脊液和其他无菌体液中主要病原菌是大肠埃希菌、CNS、屎肠球菌、肺炎克雷伯菌,鲍曼不动杆菌和铜绿假单胞菌。应防范泛耐药肠杆菌科细菌、铜绿假单胞菌和鲍曼不动杆菌对临床治疗造成严重威胁。     

BD BACTEC 9120全自动血培养仪阳性结果分析

目的评价BD BACTEC 9120全自动血培养仪阳性结果中的菌群类型和假阳性/假阴性情况。方法分析3100份临床标本中的病原菌种类和出现的假阳性/假阴性率及原因。结果BD BACTEC 9120全自动血培养仪假阳性率和假阴性率分别为6.35%和1.32%,假阳性中以白细胞增多和肿瘤患者为多。所有致假阴性菌种中,以真菌的比率最高;血培养病原菌以金黄色葡萄球菌和大肠埃希菌为多,分别为42.5%和25.5%。结论了解血培养菌群类型有助于指导临床用药,对BD BACTEC 9120血培养仪报道的阴性结果必须进行传代培养。     

全自动血液分析仪检测特殊体液有核细胞与手工法的结果比对

目的比对Sysmex XE-5000全自动血液分析仪(XE-5000)与手工法检测特殊体液有核细胞计数与分类的结果。方法100份特殊体液(脑脊液、浆膜腔积液)标本同时用XE-5000与传统的显微镜手工法进行有核细胞计数与分类。结果当有核细胞计数为31~1 000/μL时,2种方法检测有核细胞计数的结果差异无统计学意义(P>0.05),并且检测的有核细胞计数和有核细胞分类结果具有相关性(r分别为0.981和0.991,P<0.05)。结论当有核细胞计数为31~1 000/μL时,用XE-5000检测特殊体液有核细胞的准确性好、精密度高。     

Manual versus automated streaking system in clinical microbiology laboratory: Performance evaluation of Previ Isola for blood culture and body fluid samples

Background

The process of plate streaking has been automated to improve routine workflow of clinical microbiology laboratories. Although there were many evaluation reports about the inoculation of various body fluid samples, few evaluations have been reported for blood. In this study, we evaluated the performance of automated inoculating system, Previ Isola for various routine clinical samples including blood.

Methods

Blood culture, body fluid, and urine samples were collected. All samples were inoculated on both sheep blood agar plate (BAP) and MacConkey agar plate (MCK) using Previ Isola and manual method. We compared two methods in aspect of quality and quantity of cultures, and sample processing time. To ensure objective colony counting, an enumeration reading reference was made through a preliminary experiment.

Results

A total of 377 nonduplicate samples (102 blood culture, 203 urine, 72 body fluid) were collected and inoculated. The concordance rate of quality was 100%, 97.0%, and 98.6% in blood, urine, and other body fluids, respectively. In quantitative aspect, it was 98.0%, 97.0%, and 95.8%, respectively. The Previ Isola took a little longer to inoculate the specimen than manual method, but the hands‐on time decreased dramatically. The shortened hands‐on time using Previ Isola was about 6 minutes per 10 samples.

Conclusion

We demonstrated that the Previ Isola showed high concordance with the manual method in the inoculation of various body fluids, especially in blood culture sample. The use of Previ Isola in clinical microbiology laboratories is expected to save considerable time and human resources.

     

2012年中国CHINET无菌体液中分离的细菌构成和耐药性监测

目的：了解国内不同地区15所医院2012年从临床分离自脑脊液及其他无菌体液（胸水、腹水、胆汁等）的细菌构成及其对抗菌药物的耐药性。方法对国内主要地区（13所综合性医院、2所儿童医院）临床分离菌株采用纸片扩散法或自动化仪器法按统一方案进行细菌药敏试验,按CLSI 2012版标准判断结果。结果3945株非重复的细菌中,革兰阴性菌2214株,占56．1％,革兰阳性菌1731株,占43．9％。无菌体液中最常见的细菌依次为大肠埃希菌,肺炎克雷伯菌,凝固酶阴性葡萄球菌（CNS）,屎肠球菌,粪肠球菌。无菌体液标本中甲氧西林耐药金黄色葡萄球菌（金葡菌）和CNS（MRSA和MRCNS）的检出率分别为61．3％和77．2％。未发现对万古霉素、利奈唑胺耐药的葡萄球菌。无菌体液中发现1株对万古霉素耐药的粪肠球菌和1株对利奈唑胺耐药的粪肠球菌,还发现6株对万古霉素耐药的屎肠球菌。无菌体液标本中产ESBL大肠埃希菌和肺炎克雷伯菌的检出率分别为48．1％和24．4％。无菌体液标本中泛耐药鲍曼不动杆菌、肺炎克雷伯菌和铜绿假单胞菌的检出率分别为24．5％、5．5％和1．4％。结论应防范泛耐药肠杆菌科细菌和鲍曼不动杆菌对临床造成严重威胁。     

2012年度某院患者无菌体液耐药性监测的调查

目的 了解2012年度该院患者无菌体液的细菌分布和对抗菌药物的耐药性.方法 对临床送检的无菌体液标本按常规进行病原菌分离,采用Vitek2-Compact系统和ATB Express系统进行鉴定,测定抗菌药物的最低抑菌浓度（MIC）值,采用纸片扩散法（K-B）进行微生物敏感性试验.应用WHONET 5.6软件进行数据分析.结果 分离菌株556株,革兰阴性菌316株（57%）,革兰阳性菌226株（40%）.常见细菌分别为大肠埃希菌、凝固酶阴性葡萄球菌、肺炎克雷伯菌、粪肠球菌、屎肠球菌、鲍曼不动杆菌、铜绿假单胞菌.大肠埃希菌、肺炎克雷伯菌的检出率分别为53.3%和21.1%;检出9株金黄色葡萄球菌,其中,耐甲氧西林金黄色葡萄球菌（66%）;耐甲氧西林凝固酶阴性葡萄球菌（74%）.未发现对万古霉素和利奈唑胺耐药的葡萄球菌.未发现对万古霉素耐药的肠球菌属细菌.大肠埃希菌、肺炎克雷伯菌对碳青酶烯类耐药的检出率分别为3.8%和3.8%.结论 及时监测病原菌的菌群种类、分布和耐药变迁以指导临床合理、规范地使用抗菌药物.     

Comparison of BACTEC plus Aerobic/F,Anaerobic/F,Peds Plus/F,and Lytic/F media with and without fastidious organism supplement to conventional methods for culture of sterile body fluids

We compared the BACTEC 9240 continuous-read instrument using Peds Plus/F, Lytic/F, Aerobic/F, and Anaerobic/F media (Becton Dickinson Diagnostic Instrument Systems, Sparks, MD) with and without fastidious organism supplement to conventional centrifugation preparation and plating for the recovery and speed of detection of microorganisms. A total of 908 sterile body fluid specimens were collected and processed, yielding 116 (13%) positive cultures. Of the 80 isolates considered clinically significant, 48 (60%) were recovered by both the BACTEC system and conventional culture, whereas 32 (40%) were recovered by BACTEC only. No clinically significant isolates were recovered only by conventional culture methods. The time to detection for isolates recovered from both sets was faster for BACTEC. It was found that BACTEC, with or without the addition of fastidious organisms supplement, exhibited improved sensitivity for the recovery of microorganisms.     

ESP全自动血培养仪的临床应用及评价

目的　评价ESP全自动血培养仪(简称ESP)临床应用情况。方法　用ESP检测2147份血标本,观 察其阳性检出率、检出时间,并对所检出细菌的种类及阳性率进行分析。结果　2147份血标本中ESP提示阳性 322例,分离出294株细菌,阳性率为13.7%,最快检出时间为2h,24h内检出的阳性数占60.5%,48h内检出 的阳性数占83.3%,72h检出的阳性数占99%。假阳性率为0.87%。未发现假阴性。结论　应用ESP有较高 阳性检出率,缩短了培养时间,增加了检出细菌的种类,结果快速、准确。     

Evaluation of the BacT/Alert microbial detection system with FAN aerobic and FAN anaerobic bottles for culturing normally sterile body fluids other than blood

We evaluated the BacT/Alert Microbial Detection System (Organon Teknika Corporation, Durham, NC, USA) by using FAN bottles compared to conventional culture methods for the recovery of microorganisms from normally sterile body fluids other than blood and dialysates. Clinically significant pathogens were isolated from 116 (11%) of 1, 099 consecutive specimens (80 from both conventional media and FAN bottles; 23 from FAN bottles only; 13 from conventional media only). Gram-positive cocci were more likely to be recovered from FAN bottles than from conventional media (p = 0.04). Contaminants were also more likely to have grown in FAN bottles (3%) than on conventional media (1%) (p = 0.04). The mean time to detection of significant pathogens was 20.9 h using FAN bottles as compared to 30. 9 h using conventional media (p = 0.0001). These results indicate that the BacT/Alert Microbial Detection System using FAN blood culture bottles improves the yield of clinically significant Gram-positive isolates from normally sterile body fluids with a reduced time to detection.     

Blood culture examinations at a community hospital without a microbiology laboratory: using an automated blood culture system and performing a Gram stain on positive culture bottles in the institution

To elucidate the existence of microorganisms from blood culture bottles in hospitals without a microbiology laboratory, we changed the system of blood culture examinations. The Oxoid signal blood culture system and submission of all blood cultures to the clinical testing industry was used from July 2002 to December 2002 (first period). Use of the BacT/Alert system and performing of Gram stain for positive culture bottles in our institutions was conducted from January 2003 to June 2003 (latter period). A total of 210 and 193 blood cultures were processed during the first and latter periods, respectively. There were 40 (19.0%) positive cultures in the first period and 32 (16.6%) positive cultures in the latter period. The times from the specimen collection to the Gram stain result that were required were 3.8 and 1.0 days in the first period and the latter period, respectively. The times required for the final report of the blood cultures in the first period and in the latter period were 5.8 and 4.9 days, respectively. We conclude that using a continuous monitoring, automated blood culture system and performing Gram stain for positive culture bottles in institutions without microbiology laboratories may be useful for medical doctors to rapidly determine the existence of microorganisms and to begin adequate antiinfective therapy.     

XT-4000i血细胞分析仪在体液细胞计数的应用分析

目的：探讨 XT-4000i 血细胞分析仪（简称 XT-4000i）在体液细胞计数中的应用价值。方法收集2013年9月份至2014年2月份住院患者体液标本113份,将收集的标本1 h 内分别在 XT-4000i 血细胞分析仪上和手工检测红细胞（RBC）和白细胞（WBC）,并将 RBC 值分为3个浓度水平 L1：100～1000×106／L;L2：1001×106～100000×106／L;L3：＞100000×106／L, WBC 分为2个浓度水平 L1：1～50×106／L;L2：＞50×106／L 并对2种检验方法进行相关性分析。结果 XT-4000i 血细胞分析仪与手工法检测 RBC、WBC 的结果具有较高的相关性,相关系数分别为0.931、0.996、0.865、0.942和0.988。结论 XT-4000i血细胞分析仪可以在临床体液细胞计数中应用。     

实行血培养阳性危机值报告制度监督血培养全程质量控制

目的 实行血培养阳性危机值(critical value)报告制度监督血培养全程质量控制(TQC),提高血培养阳性率,快速、准确地作出病原学诊断.方法 血培养及鉴定采用美国BD9050全自动血培养仪和法国生物梅里埃公司ATBExpression自动细菌鉴定仪.结果 在2 700例血培养中阳性共326例,其中4例为污染菌,污染率为1.2%;322例为病原菌,阳性率为12%.结论 实行血培养阳性危机值报告制度使微生物检验人员、临床医生和护理人员及时沟通,减少污染率,实现血培养全程质量控制.     

临床科室常规无菌物品管理方法的改进与效果

目的发挥消毒供应中心专业优势,规范常规无菌物品在科室的安全储存管理,提高无菌物品终末环节的管理质量。方法调查常规无菌物品在科室管理环节存在的问题,进行论证分析,消毒供应中心协同护理部、感染控制科进行相关知识培训,制订常规无菌物品管理制度,细化环节工作流程,对常规无菌物品进行指导管理并持续质量控制。结果常规无菌物品在科室储存管理问题减少,常规无菌物品管理质量提高(P0.01)。结论消毒供应中心与护理部、感染控制科共同参与管理,提高了常规无菌物品在科室储存环节的管理质量,保证了无菌物品使用时的安全性、有效性。     

血培养仪阳性报警时间在血培养检出菌性质鉴别中的应用

目的 探讨血培养仪阳性报警时间(TP)对所检出的凝固酶阴性葡萄球菌(CoNS)性质鉴别中的应用价值.方法 对236例血培养检出CoNS的新生儿血培养标本TP和定量血培养(QBCs)检测结果 进行比较分析,根据QBCs、TP,并结合临床资料,判断CoNS是感染菌或污染菌.结果 TP和QBCs呈显著负相关:TP≤16 h时,QBCs＞100 CFU/mL;TP>20 h时,QBCs＜10 CFU/mL.QBCs检测结果 与患儿临床症状严重程度和抗菌药物治疗的必要性呈明显正相关:QBCs＜10 CFU/mL,即TP≥20 h,预示细菌污染;QBCs＞100 CFU/mL,即TP≤16 h,预示血液中细菌浓度较高,存在血流感染,需积极采取抗菌药物治疗.结论 TP作为判断细菌浓度的指标,也可用于鉴别血培养检出CoNS是否提示血流感染.     

人脐血间充质干细胞培养方法的比较

背景：脐血中含丰富的间充质干细胞,可以作为组织工程中一种新的种子细胞来源。目的：应用两种培养基体外培养、扩增、分离纯化脐血间充质干细胞,比较其生物学特性的差异。方法：无菌条件下采取40份足月顺产的脐血,肝素抗凝。应用Ficol 密度梯度离心法分离脐血单核细胞,随机分为两组,其中20份用MesenGro人间充质干细胞培养基进行培养,20份用DMEM培养基进行培养。对比两组梭形的间充质干细胞出现时间、细胞集落出现时间、培养时间、原代细胞数量。选用生长情况良好的原代脐血间充质干细胞,用流式细胞仪检测间充质干细胞表面特异性标记表达情况。结果与结论：MesenGro组梭形的间充质干细胞平均出现时间、细胞集落平均出现时间、原代细胞平均培养时间、原代细胞平均数量为均优于DMEM组（P 〈0.01）。流式细胞仪检测显示,培养的细胞强表达间充质干细胞表面标志CD73和CD105,阳性率99.1%,不表达造血干细胞表面标志CD45和CD34,阴性率99.3%。结果提示在培养间充质干细胞的数量、形态、生长速度和培养时间诸方面,MesenGro 人间充质干细胞培养基均优于DMEM培养基。选用MesenGro人间充质干细胞培养基可以更纯、更快、更好地从脐血中培养出表达间充质干细胞表面标志的细胞。     

3种体液细胞染色方法的效果比较

目的选择简单、快速且能反映体液细胞原有结构特征的染色最佳方法。方法比较临床较为常用的苏木精-伊红(HE)染色法、巴氏染色法及瑞-姬(WG)混合染色法对650份不同体液标本进行细胞染色效果及阳性检出率。结果 WG混合染色法操作简便快速,能使体液细胞结构最大限度地保持原有状态及特征,细胞质、细胞核及核仁之间的色差大,易于区分辨认,阳性诊断符合率为99.7%、灵敏度为79.0%、总有效率达94.0%。结论经对各种体液细胞染色观察分析,WG混合染色法效果优于HE和巴氏染色法,特适用于各级检验单位体液细胞的开展。     

两种需氧血培养系统对模拟菌血症检测能力的比较

目的比较BACTEC PLUS树脂需氧瓶与BacT／Alert FA活性炭需氧瓶对模拟菌血症的检测能力。方法选择烧伤科常用的抗生素与细菌病原体,将菌液、抗生素、新鲜无菌血注入各培养瓶内,记录5d内阳性瓶的检出时间及二者对万古霉素的吸附能力,对2种具有不同吸附剂的血培养瓶的阳性检出率及平均检出时间进行比较。结果除替考拉宁组外,BACTEC PLUS树脂需氧瓶的细菌阳性检出率和平均检出时间均优于BacT／Alert FA活性炭需氧瓶,差异有统计学意义（P〈0．001）;替考拉宁组BacT／Alert FA活性炭需氧瓶的细菌阳性检出率和平均检出时间优于BACTEC PLUS树脂需氧瓶。结论在进行血培养时,应该尽可能在抗生素使用以前或F次使用抗生素以前采血。对已经应用抗生素的患者,应选择具有中和、吸附抗生素能力的培养系统。     

直接药敏试验与常规药敏试验在血培养中的应用比较

目的评价直接药敏试验和常规药敏试验在血液细菌感染中的临床应用意义。方法随机选取2012年3月至2014年5月广州中医药大学附属新会中医院280份血培养阳性标本,对上述280份标本分别予以直接药物敏感试验和常规药敏试验,研究2种检测结果的细菌鉴定结果以及药敏结果在2种不同方法中的相关性。结果直接细菌鉴定法与常规细菌鉴定法的一致性为92.5%,2种检测方式的敏感度、药物耐药及中度敏感的符合率比较,差异无统计学意义(P0.05)。结论直接药敏试验可以缩短血培养阳性标本的报告时间,有效指导临床合理选用抗菌药物。