环境采样标本倾注平皿和平板划线两种方法接种结果对比分析

目的 探讨倾注平皿法和平板划线法接种效果的灵敏性和检出细菌的阳性率.方法 将2010年12月-2011年3月环境采样标本219份和透析用水样本84份,分别采用1 ml倾注平皿法和100μl平板划线法接种,培养48 h后判读结果,比较细菌生长标本数和阳性率.结果 消毒液、物体表面、手和透析用水标本采用倾注平皿法和平板划线法培养出的阳性率分别为0、0、17.19％、1.72％、25％和0、1.33％、15.63％、5.17％、35.71％,经t检验,仅透析用水采用两种方法接种的细菌生长数差异有统计学意义(P＜0.05),但其阳性率差异无统计学意义;其他标本采用两种方法接种培养的结果阳性率及细菌生长计数差异均无统计学意义.结论 实际工作中平板划线法简单、易行,可操作性强,可代替倾注平皿法.     

医院微生物室检验人员职业暴露与自我防护

在微生物检验工作中,完善防护设备,严格执行各种操作规程,加强微生物室工作人员自我防护意识很重要. 1 微生物室检验工作中存在的危害 1.1 病原微生物危害主要指病原微生物造成的实验室污染,由于每天要接种和处理大量具有潜在传染性的标本,这就增加了工作人员接触标本发生感染的机会.另外,在灼烧接种环时,标本飞溅、稀释和移加菌液过程中产生的气溶胶,不仅增加了工作人员的呼吸道感染机会,也污染了工作台面和地面.     

细菌培养框研制及应用

细菌培养标本根据要求接种于不同的培养基,放入培养箱中经18～24 h后,观察每个培养皿细菌的生长及菌落的形态,并进行革兰染色,这是细菌室每日必须的工作流程,为了培养出致病菌,1份标本均要选择≥2种的培养皿,培养标本多时,所种的培养皿也很多,面对培养箱中杂乱放置的培养皿,想寻找与一张化验单相对应的培养皿,颇费功夫.     

3818例临床标本高渗培养细菌的探讨

目的　加深对开展高渗培养有利于临床标本细菌 L型的检出及能弥补普通培养的不足的重要性的认识。　方法　普通培养和高渗培养。　结果　高渗培养基中细菌的检出情况 ,3 818份标本中总检出率为 3 9.86% ,其中尿高渗培养的阳性率高于血液 ;高渗培养基中检出的细菌分布情况 ,15 2 2例培养阳性标本中共检出细菌 19种 ,尿中最多为肠球菌 ,其次为金黄色葡萄球菌等 ;3 4 2份标本同时进行普通培养和高渗培养结果显示 ,高渗培养细菌检出率明显高于普通培养 ;高渗培养基中细菌 L型及细菌型的检出情况表明 ,细菌 L型只占少数 ,大部分是典型的细菌型。　结论　同时开展高渗培养可提高临床标本细菌检出率 ,减少漏诊。加强培养基质量、标本接种量及方法等方面的质控 ,有利于临床标本细菌L型的检出     

血培养阳性标本直接细菌鉴定和药敏分析

目的：对血培养阳性标本直接细菌鉴定和药敏进行分析。方法研究我院收治的发热合并全身感染患者86例,在专用接种水中混掺患者的血培养标本,并在鉴定板上接种,通过直接法鉴定,同时把阳性标本转种平板进行分离培养,根据常规法对分纯后的菌落实施上机鉴定及药敏实验,然后进行对比。结果在86例样品当中,直接法和常规法符合率：细菌鉴定总符合率为96.4%,其中革兰阴性菌95.8%,革兰阳性球菌97.1%;两种方法的细菌结果均有着较高的准确率,细菌鉴定准确率在95.7%以上。结论血培养阳性标本直接细菌鉴定和药敏实验能够准确的早期诊断菌血症患者,值得推广。     

血培养阳性标本细菌鉴定和药敏试验的临床研究

目的探讨血培养阳性标本细菌鉴定和药敏试验在临床应用中的可行性。方法选取2009年2月至2011年来我院接受菌血症治疗患者的血培养阳性标本培养液80例,均匀混于接种水中,并接种于鉴定板上依次进行生化微管法细菌鉴定和药敏试验,比较检测结果的符合率。结果 80例样品中,生物微管法细菌鉴定结果准确率均高于84.61%,χ2检验结果为P<0.05,无统计学意义;药敏试验结果的符合率均高于95.45%,χ2检验结果为P<0.05,无统计学意义。结论血培养阳性标本细菌鉴定和药敏试验可为菌血症患者进行准确的早期诊断,适于在基层医院进行推广使用。     

农村呼吸道感染的首选药物——162株痰液致病菌的药敏实验结果分析

随着抗生素的广泛使用，耐药细菌在临床上成普遍现象，合理使用抗生素显得非常重要，现将我院近年来临床疾液致病菌药敏实验结果分析如下。１材料与方法标本是我院１９９３年５月－１９９８年５月的临床标本，分离出病原菌１６２株。疾液接种于血平皿上，３５℃孵育４８ｈ...     

2011年沈阳市结核菌分离培养及影响因素的研究

目的分析2011年沈阳市肺结核细菌分离培养的情况及影响因素。方法针对结核菌分离培养，从标本收集、实验室操作、试剂培养基质控、实验室设备设施及生物安全等方面进行分析。结果通过对影响肺结核细菌分离培养因素的有效控制达到了培养阳性率大于90%，污染率小于5%的要求。结论肺结核细菌分离培养的影响因素一是实验室设备设施、试剂培养基质量等，二是人员的操作技术，业务素质，更为重要的是相应的操作规程、管理工作的严格执行。     

痢疾杆菌培养的改良方法

大便标本做痢疾杆菌培养,阳性率低,是从事细菌检验工作的同志经常遇到的一个问题。为了提高痢疾培养的阳性率,特作了增大样品量及接种环的改良,现将改良后观察情况介绍如下: 1.自制接种器用一支16号自行车条,加工成一个带柄的底边长4cm的等腰三角形,柄长6—8cm(如图1)或制成三角形缺一个腰边的形状(如图2)     

应用微菌落技术对细菌进行快速定量测定

本文利用微菌落技术建立了一种对细菌进行快速定量测定的方法，其基本方法为将待测标本定量接种于醋酸纤维素薄膜上，经３７℃５ｈ培养，染色后对标本区的微菌落作显微镜计数，用本法与平板计数法同时对大杆菌和金黄色葡萄球菌悬液进行测定，得到很好的相关性，对７２份尿液标本测定结果作统计学分析，二者差别无显著性。     

Safe inoculation of blood and bone marrow for liquid culture detection of mycobacteria

BACKGROUND: Needlestick injuries confer an unnecessary risk of occupational bloodborne infections such as human immunodeficiency virus (HIV), hepatitis B virus (HBV) and hepatitis C virus (HCV) infections. After an accidental needlestick injury, procedures for inoculation of liquid culture media for rapid detection of Mycobacterium tuberculosis complex and other mycobacteria from blood and bone marrow specimens were reviewed. AIM: To identify a safer transfer device, which could replace the ordinary syringe in inoculation of liquid culture vials. METHODS: We identified a transfer device to transfer blood or bone marrow specimens from bedside tubes into liquid culture vials. CONCLUSION: The changed procedure will reduce the risk of needlestick accidents and be of benefit to other microbiological laboratories using the same or similar inoculation techniques.     

人芽囊原虫在DMEM单相培养基中体外培养方法的建立

目的建立人芽囊原虫(Blastocystis hominis,B.h)在DMEM单相培养基中的体外连续培养方法,为进一步研究其诊断、生活史、致病机制奠定基础。方法比较不同pH值、血清种类、血清浓度及接种量等条件下虫体生长繁殖情况及影响因素。结果使用DMEM培养基、接种量大于105/管、pH7.0~8.0、10%~30%小牛血清(或人、马血清),青、链霉素及二性霉素B,于37℃条件下厌氧培养,每三、六或五天转种一次,可达到体外长期培养B.h的目的。结论使用DMEM单相培养基可用于人芽囊原虫的诊断及体外长期培养。     

解脲、人型支原体固体和液体培养基方法比较

目的：探讨泌尿生殖道支原体培养试验中假阳性、假阴性结果的原因及改进的对策，研究临床检测解脲（Uu）和人型支原体（Mh）培养的方法。方法：对208例泌尿生殖道拭子直接用固体培养分离Uu和Mh，同时接种液体培养基作为对照。观察并记录24 h、48 h、72h固体培养基长出支原体菌落的阳性例数和液体培养阳性例数，并用统计学方法分析两种方法的差异。结果：固体培养基直接分离培养结果与液体培养在不同时间段所得到的结果，阳性率无统计学差异，99%的菌落于24 h～48 h之间可辨认。结论：固体培养基直接分离Uu和Mh具有鉴别准确，选择性强和菌落特征明显的优点，与液体培养相比更适用于支原体的临床检测。     

Mouse foot-pad inoculation as an aid to the isolation of Leishmania spp. from patients

Portions of splenic or subcutaneous saline aspirates from suspected visceral or cutaneous leishmaniasis patients were inoculated into NNN media with an overlay of Schneider's medium or Schneider's medium alone for routine parasitological diagnosis. The remaining portions of the aspirates were used for preparing Giemsa-stained smears and for subcutaneous inoculation into hind foot-pads of Balb/c mice. Saline aspirates obtained from the foot-pads 2-14 d after inoculation were inoculated into Schneider's medium and examined for promastigotes. Parasite isolation was achieved from 90% of confirmed leishmaniasis patients by either culture method alone. Mouse foot-pad aspiration demonstrated parasites in 95% of all patients, and in over 80% of the confirmed cases of leishmaniasis. Combined culturing and aspirate smear examination was more efficient than foot-pad inoculation alone for the demonstration of leishmanial infection. Foot-pad aspiration does not entail killing animals and was sensitive for parasite isolation; it may be a useful short-term adjunct to existing parasite isolation methods, especially under field conditions where the risks of culture contamination may be high.     

Removal of faecal coliform in high pH ponds using rapid growth alga biomass

A simple device for an efficient waste water treatment is described. The procedure uses a massive microalga culture as a source of OH^‐, which increases the pH of the medium killing the bacterial population. In the device the alga culture remains separated from the waste water by a mesh of synthetic material or a ceramic permeable to OH^‐.     

Chinese hamster ovary cells produce an enzyme that Nicks heat-labile enterotoxin from enterotoxigenic Escherichia coli

Chinese hamster ovary (CHO) cells were examined for production of an enzyme that nicked the polypeptide chain of the heat-labile enterotoxin from enterotoxigenic Escherichia coli between the A1 and A2 fragments of its A subunit. Serum-free culture medium prepared each day after CHO cell inoculation was concentrated 100 times and its proteolytic activity for formation of the A1 fragment was examined by Western blotting with anti-LT A antibody. The A subunit was detected in culture medium on day 6 after cell inoculation, although not in media on day 1 or 3, indicating that CHO cells produced a nicking enzyme. This nicking enzyme had an optimal pH of about 7.5 and an apparent Mr. of 120,000, as seen by Superose 12 TM gel filtration with an FPLC system. The activity of this enzyme was strongly inhibited by diisopropyl fluorophosphate and phenylmethylsulfonyl fluoride, but not by p-chloromercuribenzoic acid, EDTA or ethyleneglycol bis (beta-aminoethylether)-N,N-N',N'-tetraacetic acid, suggesting that this enzyme was a serine protease. The activity was not stimulated by plasminogen or fibrin. These findings suggest that the nicking enzyme was different from proteases such as elastase, collagenase and plasminogen activator, which are probably also secreted by fibroblast-like CHO cells.     

Effect of uterine fluid from mice with IUDs,and various chemical compounds,on mouse eggs in culture

Neat uterine fluid from the IUD horn of pro-oestrous mice is slightly more toxic than control horn fluid to mouse morulae and 2-cell eggs in culture. The toxic action is completely eliminated when IUD-horn fluid is diluted with culture medium. The effects of various chemical compounds that may be elevated in the IUD horn were also tested in the egg culture system. Only a few of the compounds were found to be toxic. The significance of these results in relation to the contraceptive effect of the device is discussed.     

Effect of dietary fiber on a guinea pig intestinal anaerobe, Bacteroides ovatus

The purpose of this study was to determine the effect of fibrous plant components on the growth of intestinal bacteria. An anaerobe was isolated from the guinea pig cecum and identified as Bacteroides ovatus. The organism was grown anaerobically in two types of media and shown to require hemin or protoporphyrin IX. Treatment of the media with water-insoluble fractions of alfalfa, cabbage, spinach or wheat bran inhibited growth of the culture. Inhibition occurred whether the residue remained in the medium during culture growth or was removed before inoculation. A chelating resin also removed an essential component of the medium. Both treatments depleted the medium of hemin and addition of hemin restored growth and acid production. Treatment of the water-insoluble residues of alfalfa or cabbage with dilute NaOH decreased their inhibitory effects. This suggests that plant cell wall components possess unique and labile chemical properties. Dietary fiber may inhibit the growth of some anaerobic species in the lower intestinal tract by making hemin unavailable.     

Contribution of PCR in the diagnosis of extrapulmonary tuberculosis

Extrapulmonary tuberculosis is usually more difficult to diagnose than pulmonary tuberculosis. It often involves inaccessible sites and it is paucibacillary. OBJECTIVE AND METHOD: In this study, we tried to analyze the performance of various bacteriological methods used to diagnose 51 cases of extrapulmonary tuberculosis in an infectious diseases ward. RESULTS AND COMMENTS: The culture was positive for 55% of patients. The new amplification methods used were very disappointing for the testing of nonrespiratory samples. The sensitivity of PCR was 32% compared to that of diagnostic culture which remains the most sensitive reference method. Liquid media allow for rapid growth and limit the contaminations. The combined inoculation of liquid and solid medium increases the sensitivity and helps to identify the pathogen.     

Salmonella isolation with Rappaport's medium after pre-enrichment in buffered peptone water using a series of inoculum ratios

The ability of malachite green/magnesium chloride broth (Rappaport's medium) to isolate salmonellas from 25 ml quantities of sewage-polluted natural water was investigated. Samples were first pre-enriched in buffered peptone water and varying volumes of inoculum from the pre-enrichment culture were inoculated into Rappaport's broth. Inoculum ratios in the range 1:2000 to 1:10 were examined. The inoculum ratio denotes the ratio of the volume of inoculum to the volume of fluid medium into which it is introduced. Optimum results were obtained with the 1:2000 ratio, although the salmonella isolation rate was only slightly less with the 1:500 and 1:100 ratios. The 1:2000 inoculum ratio was obtained with a graduated loop holding approximately 0.005 ml of fluid. Use of a loop for inoculation has advantages in speed of performance and safety of manipulation.