测试片法对医疗机构环境物体表面细菌检测效果评价

目的比较菌落总数测试片和倾注平板计数法在医疗环境物体表面细菌定量检测效果,找出两种测试方法之间的相关性。方法分别用测试片法和倾注平板计数法对某三甲医院II、III类环境物体表面细菌污染检测进行比对试验,并对两种方法 48 h的培养结果进行分析对比。结果配对t检验结果表明,菌落总数测试片法和平板计数法检测菌落总数结果基本一致,无显著统计学意义(P0.05),且两种检测菌落总数方法的结果高度相关(r≈1)。菌落总数测试片法检测结果的RSD(相对标准偏差)略低于平板计数法,两者基本一致。结论测试片法在物体表面菌落总数检测中,一致性和精密度与传统活菌计数方法结果基本一致。检测过程中,测试片法具有培养箱体积小,结果清晰易读,省却繁琐的实验室准备工作等优势,缺点则为价格较贵。     

Petrifilm菌数测试片消毒质量检测中的应用研究

摘要 目的 研究一种菌数测试片在消毒质量监测中的应用价值，探索现场卫生学质量简易监测方法。方法 采用Petrifilm细菌总数测试片法对医疗机构物体表面消毒质量监测应用效果，同时与传统采样检测法作平行比较。结果 两种培养方法培养后的存活菌数基本一致，实验室模拟测试结果两组方法计数菌落数也无差别。结论 Petrifilm细菌总数测试片法可以应用于基层医疗机构的消毒质量检测工作。     

纸片法与培养发酵法检测茶具大肠菌群污染结果的比较

经比较，以大肠菌群快速检验纸片法与培养发酵法检测茶具大肠菌群的结果无显著性差异（Ｐ＞０．０５）．同样，用纸片法检测大肠菌群单项指标亦可取代大肠菌群与细菌总数的双项指标综合评价．     

浦城县电子游艺厅细菌污染检测

２０００年５月,对浦城县城区２５户游艺厅内空气用琼脂平板沉降１０ ｍｉｎ采样,检测细菌总数;对１２５台游艺机台面、按钮及１００名游艺者手用棉拭涂抹采样,检测细菌总数,用快速检验试纸片采样、检测大肠菌群。结果,厅内空气细菌总数超标（＞４０ ｃｆｕ／平板）率为 ８４．０％（４２／５０）;游艺机台面、按钮细菌总数超标（≥１０ｃｆｕ／ｃｍ２）率为８７．２％（１０９／１２５）,大肠菌群检出率为２７．２％（３４／１２５）;游艺者手细菌总数超标（≥１０ｃｆｕ／ｃｍ２）率为８２．０％（８２／１００）,大肠菌群检出…     

纸片法与增减发酵法检测茶具大肠菌群污染结果的比较

经比较，以大肠菌群快速检验纸片法与培养发酵法检测其茶具大肠菌嫩的结果无显著性差异（Ｐ〉０．０５）。同样，用纸片法检测大肠菌单项指料可取代大肠功笋与细菌总数的双项指标综合评价。     

包头市公用茶具消毒效果检测

１９９６年，对包头市１６家旅店、８家娱乐场所的消毒后公用茶具进行了细菌污染检测。对旅店茶具，于消毒后存放２４ｈ与７２ｈ，娱乐场所茶具于散场时消毒或开场前消毒后，用棉拭涂抹采样，检测细菌总数。另用大肠菌群快速检验试纸贴于茶具内外缘各２片，底部１片，３０...     

晋中市榆次区托幼机构消毒质量调查

<正> 为加强托幼机构消毒管理工作,于1998～2001年每年4～6月份对全区208所托幼机构进行消毒监测。 检测方法,对教室、卧室空气,是将普通营养琼脂平板(直径9 cm)放在室内各采样点暴露5 min采样,盖好置37℃恒温箱培养48 h,计数菌落数。对物体表面(桌面、毛巾、玩具、门把手等)采样是用浸有无菌生理盐水棉拭子在5 cm×5 cm被检物体表面涂抹后,将棉拭子放入10 ml无菌生理盐水试管中检测细菌总数。对餐饮具用大肠菌群快速检测纸检测大肠菌群。     

纸片培养法与平板培养法监测牙科水路细菌污染的比较研究

目的研究纸片培养法监测牙科综合治疗台水路(DUWLs)菌落总数的可行性。方法随机检测牙科综合治疗台水路60例样本,用纸片培养法和我国GB/T5750.12-2006《生活饮用水标准检验方法》中规定的平板培养发进行培养,培养温度均为36±1℃,培养48 h,计数菌落总数及水样合格率。结果 60处DUWLs水样菌落总数合格率为36.7%,纸片法与平板法监测DUWLs水样合格率分别为33.3%和36.7%,合格率无统计学差异(χ2=0.25,P0.05)。两种方法检测60例水样菌落总数差异无统计学意义(z=-0.990,P0.05)。结论两种检测方法监测牙科水路菌落总数的结果基本一致,但纸片法操作更为简单,可用于牙科水路的日常监测。     

纸片法在托幼机构环境菌落总数检测中的应用研究

摘要 目的比较纸片法与传统平皿法对托幼机构环境菌落总数检测的效果差异,分析两者相关性,以寻找一种简便、快捷、准确的检测方法。方法 分别采用纸片法和平皿法对托幼机构工作人员手、物体表面、室内空气等样本进行检测,用纸片直接接触法和棉签涂抹法对相同物体表面采样,同时用纸片法暴露不同的时间检测室内空气沉降菌,用配对t检验分析不同检测和采样方法及暴露时间之间的结果差异。结果 纸片法与平皿法在检测工作人员手和物体表面样品菌落总数时基本一致,差异无统计学意义(P>0.05),用纸片直接接触法对物体表面采样的检测结果明显高于棉签涂抹法,两者之间差异有显著性(P<0.05)。纸片法与平皿法在检测空气菌落总数方面48 h结果无明显差异,但24 h纸片菌落计数结果明显高于平皿。纸片暴露法采样3、5、10和15 min的计数结果逐渐降低,10 min时的检测结果最接近平皿法。结论 纸片法在检测环境菌落总数方面与平皿法相关性、准确性较高,且简便、快捷,避免了大量前期准备和后期清洁消毒处理工作,有一定应用和推广价值。     

荆门市公立与私立托幼机构细菌污染调查比较

１９９８年５～７月，对荆门市城区２１所公立及１５所私立托幼机构的细菌污染状况进行了调查比较。室内空气采用普通营养琼脂平板沉降５ ｍｉｎ采样，人员手与物体表面用浸有无菌生理盐水棉拭涂抹采样，其中桌子与其它台面借助 ５ ｃｍ × ５ ｃｍ规格板涂抹采样，毛巾、玩具用无菌滤纸（５ ｃｍ × ５ ｃｍ）粘贴采样，检测细菌总数。餐饮具用快速试纸检测大肠菌群。结果的判定，以室内空气细菌总数≤５００ ｃｆｕ／ ｍ３，物体表面或手细菌总数≤ １５ ｃｆｕ／ｃｍ２为合格，餐饮具以检不出大肠菌群为合格。 结果，私立托幼机构各项细…     

两种空气微生物采样器采样方法的研究

<正> 为探讨空气微生物的采样方法,以能更好地进行污染情况调查或消毒效果测定,我们对国产的JWI-1型与MF-45型两种采样器的采样方法进行了研究。现将结果报告于下。     

Evaluation of an automated microbiologic blood culture device for detection of bacteria in platelet components

BACKGROUND: Automated culture methods have been used by several investigators to detect bacterial contamination of cellular blood components. We investigated several factors affecting detection by automated culture of bacteria in platelet concentrates (PCs).These factors included the initial contamination level in PCs, the PC sample volume, the PC sample time, and the white cell level in relation to bacteria levels in the PCs. STUDY DESIGN AND METHODS: Staphylococcus epidermidis or Escherichia coli was inoculated into freshly prepared PCs or white cell-reduced PCs to yield colony-forming unit (CFU) levels of 10, 1, or 0.1 per mL. At the time of inoculation (t=0) and at t=6, t=24, and t=48 hours, 0.5, 1.0, and 2.0 mL samples of the contaminated PCs were transferred into culture bottles. The presence of bacteria in the culture bottles was subsequently monitored by an automated blood culturing instrument. Bacteria levels in the PC at the time of first automated culture detection were determined by quantitative plating. RESULTS: E. coli was detected in 92 percent of experiments when 1.0- or 2.0-mL samples were taken at t=6 hours. At t=24 hours, 100-percent detection was observed with all tested inoculation volumes; however, by that time,>10(7) CFU per mL of bacteria were present in every PC. For S. epidermidis, 89 percent and 83 percent of contaminated PCs were detected with a t=24 hour sampling time and 2.0- or 1.0-mL sampling volume. Seven of 36 PCs with a 2.0-mL sampling volume and 10 of 36 PCs with a 1.0-mL sampling volume contained>10(6) CFU per mL of S. epidermidis at the time of first detection. CONCLUSION: Data from this preliminary evaluation suggest that sampling times of 24 hours or more would be necessary to provide confidence in detection of E. coli or S. epidermidis in PCs using this culture method.     

Quantitative detection of Escherichia coli from urine of patients with bacteriuria by real-time PCR.

INTRODUCTION: Compared with the classical urine culture method, PCR is more rapid, and can detect smaller numbers of bacteria, however it is inferior for quantification. Because of the lack of quantification in routine PCR, the meaning of a positive PCR test result has not been validated for all infections. We report on the development of a novel quantitative detection system for the urinary tract infection (UTI) Escherichia coli using real-time PCR. PATIENTS: We enrolled 200 patients with suspected bacteriuria. METHODS: The gene encoding the universal stress protein (uspA) was found to be highly specific for E. coli. We quantified the copy numbers of E. coli in the urine of patients with UTI by using a real-time PCR assay (the TaqMan system) targeting uspA genes in genomic DNAs isolated from urine samples (n=200). To evaluate the feasibility of this method, the results were compared with those of a standard urine culture. RESULTS: The incidence of positive urine cultures was 75% (150 of 200), and various doses of E. coli were detected in 84 of 150 specimens. The real-time PCR method also detected 84 cases of urinary infections of E. coli in the same specimens. Furthermore, the result of the quantification of E. coli using real-time PCR strongly correlated (r2=0.925) with the result of urine culture. CONCLUSION: Our results suggest that using quantitative-PCR means a faster and simpler diagnosis of E. coli urinary infection can be made compared with the traditional urine culture method.     

Near-patient testing for infection using urinalysis and immuno-chromatography strips.

Urinary tract infections require costly confirmatory tests such as a urine culture to establish the diagnosis. Elimination of the culture step would save resources; diagnosis and treatment could begin in hours rather than days. We tested a new dip-and-read strip that uses immuno-chromatography (IC) to detect infectious agents in urine. We used a goat-derived polyclonal antibody with reactivity to the cell-wall proteins of Escherichia coli (E. coli). Fluorescein linked to the anti-E. coli antibody served to trap the bacteria on a strip coated with an anti-fluorescein mouse antibody. Blue latex particles were linked to anti-E. coli antibodies by standard methods and were used for detection of E. coli. We found that the combination of leukocyte esterase and nitrite dipsticks gave negative predictive values of 93% for culture-negative urines, i.e., there were very few false-negative results. Using the same dipsticks on culture-positive specimens, the positive predictive values were unacceptably low; we obtained too many false-positive values. By contrast, the IC strips gave negative predictive values of 89%. The major advantage of the IC strips is that the positive predictive values were higher, i.e., there were fewer false-positive results. The combined use of both IC strips and urinalysis dipsticks offers the best strategy for diagnosing infection with dipsticks. The IC strip test could reduce the necessity of a urine culture in patients with suspected infections and provide rapid point-of-care testing.     

微菌落技术用于细菌培养计数初探

对微菌落方法，以聚碳酸酯微孔滤膜作载体，结合显微计数，用于活菌培养计数。该法于３７℃培养３５～５０ｈ检测的结果与常规琼脂倾注平板法培养２４～４８ｈ的结果无显著性差异     

ATP生物荧光技术快速测定细菌总数的应用研究

目的研究ATP生物荧光技术快速测定细菌总数的应用。方法通过生物荧光检测系统与其配套的试剂盒对细菌悬液进行定量检测,同时与平板活菌计数培养法作平行比较。结果经对金黄色葡萄球菌、大肠杆菌和绿脓杆菌菌悬液检测结果显示,三种细菌菌悬液检出菌数的对数值与ATP值的对数值之间呈线性关系。金黄色葡萄球菌、大肠杆菌和绿脓杆菌回归方程依次为Y=1.209+0.744X(R2=0.942),Y=0.885+0.831X(R2=0.961)和Y=0.955+0.953X(R2=0.983)。ATP生物荧光法适用于检测含菌量<107cfu/ml的菌悬液。结论 ATP生物荧光法可快速地评价受细菌污染的液体中细菌总数,但溶液中细菌含量应控制在<107cfu/ml。     

Algorithm for the identification of bacterial pathogens in positive blood cultures by real-time LightCycler polymerase chain reaction (PCR) with sequence-specific probes

We developed real-time polymerase chain reaction (PCR) assays for rapid detection of the most common and clinically relevant bacteria in positive blood culture bottles, including Staphylococcus spp., S. epidermidis, S. aureus, Enterococcus spp. (including differentiation of E. faecalis and E. faecium), Streptococcus spp., Streptococcus agalactiae, Enterobacteriaceae, E. coli, Pseudomonas aeruginosa, Stenotrophomonas maltophilia, Acinetobacter spp., Bacteroides spp., Haemophilus influenzae, and Neisseria meningitidis. A total of 507 positive blood cultures were investigated according to a specific PCR algorithm based on the microscopic result of the blood culture, and the PCR results were compared to the culture results. Apart from-cross reactions between E. coli and Chryseomonas luteola and Enterococcus faecium and E. durans, the PCR assay correctly identified all bacteria in the blood cultures and did not show any false-positive results. Regarding blood cultures positive with a single species of bacteria (n = 474), 98.3% of all bacteria were correctly detected by the PCR algorithm within a few hours. However, in mixed infections, the sensitivity was lower. The PCR algorithm is well suited for rapid identification of the most common bacteria in positive blood cultures.     

Detection of EspB using reversed passive latex agglutination: application to determination of enteropathogenic Escherichia coli

We developed a new practical method to identify enteropathogenic Escherichia coli (EPEC) by detecting the pathogenic factor, EspB. E. coli were cultured in Dulbecco's Modification of Eagle's Medium (DMEM), and EspB was detected in the culture supernatant by reversed passive latex agglutination (RPLA). All 63 E. coli strains harboring the eaeA gene encoding intimin were positive for RPLA, and all 25 strains without the eaeA gene were negative. Among these 63 eaeA-positive strains, 38 Shiga toxin-producing E. coli (STEC) produced Shiga toxin (Stx) under the same culture conditions (DMEM). Subtypes of EspB alpha, beta and gamma were antigenically cross-reactive to each other as determined by RPLA and Western blotting. A kit for Stx detection (RPLA) is commercially available and therefore this RPLA for detection of EspB could be a practical method to define EPEC in both clinical laboratories and the field.     

透析用水的细菌培养方法比较

目的比较几种透析用水细菌培养方法的敏感性,寻找其中方便、快速、敏感性高的方法。方法在北京市某三甲医院血透中心随机采样透析用水共85份,分别用4种方法进行细菌培养:①哥伦比亚血琼脂平板培养基37癈培养48h、②TSA培养基37癈培养48h、③R2A培养基37癈培养48h或④20癈培养7d,同时设立阳性及阴性对照组。结果 1.R2A培养基20癈培养7d细菌检出率最高;37癈培养48h细菌检出率降低,但均高于TSA培养基和血琼脂平板培养基。2.EBPG标准方法与选用R2A培养基37癈培养48h的方法具有较好的一致性,可以互相替代。结论我们建议采用EBPG建议的透析用水细菌培养方法,或者通过适当提高温度、缩短时间来改进EBPG建议的方法从而更方便临床使用。     

肺炎支原体快速检测方法的建立及应用评价

目的制备抗肺炎支原体(MP)的特异性单克隆抗体,利用单克隆抗体建立MP胶体金快速检测方法。方法采集急性呼吸道感染患儿咽拭子标本,分离培养并鉴定MP,纯化MP抗原,制备抗MP单抗,利用单抗建立MP胶体金检测试纸条。取疑似MP感染患者的咽拭子标本,分别使用荧光定量PCR法和胶体金快速检测法对其进行检测,观察并统计检查结果。结果成功分离培养出MP菌株,灭活并纯化出MP抗原,利用该抗原免疫小鼠,通过杂交瘤技术制备出抗MP单克隆抗体19株,从中选出效价高、特异性较好的两株单克隆抗体(MP-5和MP-19)作为原材料制备MP胶体金快速检测试纸条。胶体金试纸条最低检测限为20ng。分别采用胶体金试纸条和荧光定量PCR法对临床标本进行检测,结果显示,本研究制备的MP胶体金快速检测法的灵敏度为88.2%,特异度为82.6%。结论制备出抗MP的特异性单克隆抗体,已初步建立MP胶体金快速检测法,并为临床MP感染患者的快速诊断提供帮助。