肺炎支原体液体培养法检测的实验评价

目的比较液体培养法和固体培养法平行检测肺炎支原体结果的一致性;评价液体培养法检测肺炎支原体的可靠性。方法采用液体培养基和固体琼脂培养基平行检测1 648份临床标本的肺炎支原体,比较同一份标本在2种培养基上的检测结果。结果液体培养法阳性296例,阳性率为18%;固体培养法阳性244例,阳性率为14.8%;液体培养法阳性而固体培养法阴性57例;固体培养阳性而液体培养法为阴性5例。2种方法的阳性检出率比较差异有统计学意义(P<0.05)。结论肺炎支原体快速液体培养法与固体培养有较好的一致性,具有方便、简单、准确且可以用于早期检测等优点,适合临床大批量标本筛查。需结合患者临床症状等排除真菌和耐药菌造成的假阳性。     

对两种新生隐球菌选择性培养基的比较研究孙钒*

比较鸟籽琼脂 (GASA ,Guizotiaabyssinicaseedagar)和咖啡酸玉米琼脂 (CACA ,Caffeicacidcommealagar)对新生变种和格特变种的培养效果 ,再同时用两种培养基分离鸽粪和澳洲赤桉标本中的新生隐球菌。结果表明 ,CACA对新生隐球菌的培养和选择性分离效果与GASA相同 ,能够用于新生隐球菌的选择性分离。     

幽门弯曲菌MLE无血培养基的研究

用MLE无血培养基与MHC(M-H血巧克力琼脂)培养基同时进行已知幽门弯曲菌(Cp)的培养及胃粘膜活检标本中Cp的分离试验。结果表明,16株已知菌在MLE培养基上生长的菌落比MHC上丰茂,特点明显,两者的菌体形态及生化反应一致。200份胃粘膜活检标本用MLE培养基分离阳性者114例,阳性率为57.0％,用MHC分离阳性者101例,阳性率为50.5％,符合率为86.5％,分离阳性率MLE高于MHC(p<0.05)。我们认为MLE无血培养基可代替含血培养基用于培养分离Cp。     

国家标准测定食品细菌总数培养基的改进研究

应用我国国家标准营养琼脂（GB4789 2－94简称NA）与美国食品药品管理局（FDA）标准平析以（简称SA）两种培养基对动植物食品中细菌总数进行检测对比,结果表明FDA标准平板比GＢ营养琼脂效果较好,前比后的检出率高出２３．９％,且菌落大而明显,为此,对这两种培养基进行优化筛选试验,并优选出Ｃ８培养基,扩大试验结果表明C8培养基的检出率较GB营养琼脂及FDA标准平板分别高出35．8％和9．5％。     

858例血感染患者病原菌分布及耐药情况

目的:通过对某地区中心医院收集的临床血感染患者感染病原菌的分析,了解该地区血感染患者病原菌构成、分布及耐药特点,为临床治疗提供参考和指导。方法:收集2012年6月至2013年8月期间在某院就诊的858例血感染患者血液标本,采用BACTEC9050全自动血培养仪培养,采用VITEK 2 Compack系统和K-B琼脂纸片扩散法对阳性标本进行菌种鉴定和药敏检测。结果:血培养结果显示,在858份血培养标本中共检出阳性标本109份,每份标本都只检出一种病菌,总检出率为12.7%,革兰阳性菌占64.22%(70/109),革兰阴性菌占33.03%(36/109),真菌占0.35%%(3/109);药物敏感试验结果显示:葡萄球菌对青霉素、红霉素和复方新诺明耐药率40%;肠杆菌科细菌对氨苄西林和氯霉素耐药率40%;非发酵菌科细菌对氨苄西林,头孢他啶,头孢噻肟和氯霉素耐药率40%。结论:目前本地区临床血感染患者革兰阳性菌感染率高,以金黄色葡萄球菌和凝固酶阴性葡萄球菌为主,治疗可以首选糖肽类抗菌药物;革兰阴性菌以大肠杆菌和绿脓杆菌为主,对氨苄西林、氯霉素耐药率高,大肠杆菌对头孢类抗生素的耐受较绿脓杆菌低,两种细菌感染治疗可以考虑选择单环-内酰胺类抗生素。及时准确的血培养结果及药敏试验可为临床合理选择抗菌药物提供重要依据。     

香石竹的叶片培养及植株再生

１植物名称香石竹（Ｄｉａｎｔｈｕｓｃａｒｙｏｐｈｙ－ｌｌｕｓ）,别名康乃馨。２材料类别无菌苗叶片。３培养条件以ＭＳ为基本培养基。分化培养基附加：（１）６－ＢＡ１．０ｍｐ·Ｌ－１（单位下同）＋ＮＡＡ０．３;（２）６－ＢＡ１．０＋ＮＡＡ０．１;（３）６－ＢＡ１．０＋ＮＡＡ０．０５。增殖培养基附加６－ＢＡ０．５＋ＮＡＡ０．１。分化和增殖培养基均加蔗糖３％、琼脂０．７％,ｐＨ５．８。生根培养基为１／２ＭＳ附加ＮＡＡ０．１,蔗糖２％,琼脂０．６％,ｐＨ５．８。培养温度为（２５±１）℃,光照１２ｈ·ｄ－１,…     

玉香梨的组织培养与快速繁殖

１植物名称沙梨品种玉香梨（Ｐｙｒｕｓｐｙｒｉｆｏｌｉａ　ｃｖ．　ｙｕｘｉａｎｇ）。２材料类别侧芽。３培养条件（１）丛芽诱导培养基：１／２ＭＳ大量元素＋ＭＳ微量元素＋铁盐＋有机成分＋６－ＢＡ２ｍｇ·Ｌ－１（单位下同）＋ＮＡＡ０．２＋ＧＡ３２＋３％蔗糖＋０．８％琼脂,固体培养;（２）增殖培养基：１／２ＭＳ＋ＢＡ１．５＋ＮＡＡ０．２＋ＧＡ３１＋３％蔗糖＋０．８％琼脂,固体培养;（３）生根培养基：１／２ＭＳ＋ＩＢＡ１０＋１．５％蔗糖＋０．６％琼脂,固体培养８ｄ后转入不含任何激素的同类培养基中的二步生根法…     

同位素与生物素标记的HCMV DNA探针检测婴儿肝炎综合症血和尿标本的比较

用~(32)P和生物素标记的克隆化的人巨细胞病毒(HCMV)AD169株DNA片段作探针,采用DNA-DNA斑点杂交法,对照检测了27例婴儿肝炎综合症患者(血清学检测为非甲非乙型肝炎者)临床血、尿标本的HCMV-DNA。其中16份血标本呈阳性,占59％;9份尿标本呈阳性,占33％。初步结果表明,血标本中HCMV DNA检出率比尿标本检出率高26％。标记的~(32)P探针可检测10pg同源DNA,生物素探针可检测50Pg同源DNA,均不与其它疱疹病毒及未感染的人胚肺细胞DNA杂交。将其中26份血标本的HCMV DNA杂交结果与抗HCMV IgM ELISA检测结果相比较,符合率为65％。     

应用分值计算法优选SS琼脂配方的研究

用分值计算法对四批ＳＳ琼脂质量检测,分值均小于质控标准分值５６．１２５。主要问题是抑制大肠杆菌（Ｅ.ｃｏｌｉ）生长和促鼠伤寒沙门氏菌（Ｓｔｙｐｈｉｍｕｒｉｕｍ）,痢疾志贺氏菌（Ｓ．Ｄｙｓｅｎｉｃｒｉａｅ）生长的能力不够。调整ＳＳ琼脂配方中各试剂的用量进行筛选,结果表明：０．５％胆盐抑制大肠杆菌（Ｅ/ｃｏｌｉ）能力达到分值质控要求,但对鼠伤寒沙门氏菌（Ｓｔｙｐｈｉｍｕｒｉｕｍ）和痢疾志贺氏菌（Ｓ．Ｄｙｓｅｎｉｃｒｉａｅ）的生     

非洲菊的组织培养与快速繁殖

１植物名称非洲菊（Ｇｅｒｂｅｒａｊａｍｅｓｏｎｉｉ）,又名扶郎花。２材料类别无菌种子萌发的幼苗。３培养条件（１）种子萌发培养基：１／２ＭＳ＋０．７％琼脂（或脱脂棉）;（２）诱导分化培养基：ＭＳ＋６－ＢＡ１．５ｍｐ·Ｌ－１（单位下同）＋ＩＢＡ０．２;（３）芽增殖培养基：ＭＳ＋６－ＢＡ０．５;（４）生根培养基：ＭＳ＋ＮＡＡ２。上述（２）、（３）、（４）培养基内琼脂均为０．７％,蔗糖３％,ｐＨ５．８～６．０。培养条件为室温和自然光照,种子萌发前为暗培养。４生长与分化情况４．１萌发种子用纱布袋包装,无菌…     

Comparison of various culture methods (Skirrow medium, a blood-free medium and a filtration system enriched in Bolton and Preston broths) for isolation of Campylobacter spp. from raw meat samples

We compared six procedures and investigated the optimal method for isolation of Campylobacter spp. from raw meat samples. Ninety-nine meat samples were enriched in Bolton broth and Preston broth, followed by plating on Skirrow, mCCDA, and blood agar (a membrane filter on its surface) media, respectively. Thirty-nine of 99 samples were positive and 71 Campylobacter were isolated by one or more methods. More than one species of Campylobacter were obtained in 8 (20.5 %) of 39 positive samples and two genotypes were yielded on the same medium (11 samples, 28.2 %) by pulsed-field gel electrophoresis (PFGE) genotyping. Enrichment by Preston broth was significantly better than by Bolton broth (P?<?0.05). Moreover, the latter failed to detect Campylobacter jejuni strains. Skirrow medium was significantly less efficient than mCCDA medium and membrane filtration method (P?<?0.05). Overall, the combination of PC (primary enrichment in Preston broth, followed by selective enrichment on mCCDA agar), PF (primary enrichment in Preston broth, followed by membrane filtration culture onto blood agar), and BF (primary enrichment in Bolton broth, followed by membrane filtration culture onto blood agar) methods provided the optimum isolation rate of Campylobacter spp.     

Broth versus solid agar culture of swab samples of cadaveric allograft musculoskeletal tissue

As part of the donor assessment protocol, bioburden assessment must be performed on allograft musculoskeletal tissue samples collected at the time of tissue retrieval. Swab samples of musculoskeletal tissue allografts from cadaveric donors are received at the microbiology department of the South Eastern Area Laboratory Services (Australia) to determine the presence of bacteria and fungi. This study will review the isolation rate of organisms from solid agar and broth culture of swab samples of cadaveric allograft musculoskeletal tissue over a 6-year period, 2006–2011. Swabs were inoculated onto horse blood agar (anaerobic, 35 °C) and chocolate agar (CO₂, 35 °C) and then placed into a cooked meat broth (aerobic, 35 °C). A total of 1,912 swabs from 389 donors were received during the study period. 557 (29.1 %) swabs were culture positive with the isolation of 713 organisms, 249 (34.9 %) from solid agar culture and an additional 464 (65.1 %) from broth culture only. This study has shown that the broth culture of cadaveric allograft musculoskeletal swab samples recovered a greater amount of organisms than solid agar culture. Isolates such as Clostridium species and Staphylococcus aureus would not have been isolated from solid agar culture alone. Broth culture is an essential part of the bioburden assessment protocol of swab samples of cadaveric allograft musculoskeletal tissue in this laboratory.     

Improving the success of culturing Helicobacter pylori from gastric biopsies

Factors influencing the successful isolation of Helicobacter pylori from human gastric biopsies were studied. Within 24 h, each of the gastric biopsies was inoculated onto chocolate blood agar media and incubated for up to 2 weeks. Among 63 (70%) culture positive cases in 90 patients, 58 (64%) cases were culture positive for both specimens, while five (6%) cases were culture positive in only one biopsy. Of the 63 positive cultures, 51 H. pylori strains (81%) grew on both media with and without antibiotics. Eight strains (13%) grew only on medium without antibiotics, while four isolates (6%) were obtained only from medium with antibiotics. These results support the previous histological observation of patchy colonization of H. pylori in the stomach. The success rate for culture of H. pylori from gastric biopsies increased when two biopsies were taken and inoculated on chocolate blood agar media with and without antibiotics.     

人体肠道细菌寡营养培养组条件的优化研究

【目的】探讨寡营养对人体肠道细菌培养组的条件。【方法】通过稀释富集培养基、固体平板和增菌肉汤培养基成分获得寡营养培养基。对健康人粪便样本分别用原液(0)、5、10、20、30和40倍稀释的富集培养基(添加羊血和瘤胃液的血培养瓶)连续增菌,在不同时间点(第0、3、6、9、15、27、30天)吸取增菌液,用YCFA (yeast casitone fatty acid)固体培养平板分离菌落;用YCFA增菌肉汤增菌后再次挑取单菌落,利用基质辅助激光解吸/电离飞行时间(matrix-assisted laser desorption/ionization time-of-flight mass spectrometry,MALDI-TOF)质谱和16S rRNA基因测序鉴定菌株。通过比较上述6种寡营养条件分离肠道菌群的效果,选取富集培养基原液、稀释10倍和30倍这3 种条件下分离效果较好的富集条件,与同样稀释倍数条件的固体平板和增菌肉汤分别组合成9种培养基条件,进一步优化肠道菌群的培养组条件。【结果】在6种寡营养富集培养基中,未稀释(原液)、10 倍和30倍稀释的富集培养基分离细菌的种类比其他...     

An easy method for detection of nasopharyngeal carriage of multiple Streptococcus pneumoniae serotypes

In this paper, a simplified method for detection of pneumococcal carriage and for revealing the presence of several serotypes in a nasopharyngeal sample is evaluated. Enrichment broth was used for transportation and for the initial culturing of samples. All specimens were examined directly by the capsular reaction test for the presence of any of the 91 known pneumococcal serotypes. Sub-culturing on blood agar was used for isolation of the pneumococcal strains detected in the primary broth culture. A total of 693 nasopharyngeal swabs were obtained among children, their parents and employees in day care centres. Pneumococci were observed in 363 samples and 36 of these (9.9%) contained more than one serotype (multiple carriages). Two persons carried 3 different serotypes simultaneously. A significant increase in the positive sampling rate (5.8%) was achieved by using the simplified method compared to conventional streaking of the swabs directly on blood agar (p<0.0001).     

Cell surface hydrophobicity and charge of Staphylococcus aureus and coagulase-negative staphylococci from bovine mastitis

The effects of seven growth media on cell surface hydrophobicity of a collection of Staphylococcus aureus and coagulase-negative staphylococci isolated from bovine mastitis were compared in the salt-aggregation test. Thirty-three per cent of Staph. aureus strains showed extremely high cell surface hydrophobicity (auto-aggregated) and 28% were moderately hydrophobic while 26% were hydrophilic after growth on horse blood agar at 37°C for 18 h. There were great variations in the proportion and degree of the hydrophobicity depending on the medium used. Cultivations on/in capsule-inducing media caused a shift from a high to a low degree of hydrophobicity, although a microscopically detectable capsule or slime layer was seen in only one strain. This strain and encapsulated reference strains had a hydrophilic cell surface and migrated faster in free zone electrophoresis than cells of unencapsulated strains. Cells of strains grown on staphylococcus medium 110 agar migrated faster than those grown on horse blood agar regardless of their capsule production. Coagulase-negative staphylococci showed uniformly hydrophilic cell surface after cultivation on horse blood agar, but not when grown in tryptic soy broth or proteose peptone broth. It was concluded that most of the Staph. aureus strains from bovine mastitis under a variety of growth conditions in stationary phase culture constantly expressed hydrophobic cell surface.     

Comparison of microbiological culture methods in screening allograft tissue. Swab versus nutrient broth

In order to reduce the risk of transmission of infectious diseases through transplantation of tissue allografts, one should examine tissues for the presence of microorganisms. However, there are no detailed tissue banking guidelines describing the culture method or incubation time to be used. Therefore, we compared two culture methods--blood agar plate versus Wilkins Chalgren broth--and three incubation times--2, 7 or 14 days for their performance. The ultimate aim is to use the optimal setting as standard operating procedure (SOP) for tissue allograft cultures. From 70 consecutive donors, 919 tissue samples were taken. All 919 tissue samples were incubated on blood agar as well as in Wilkins Chalgren broth for 7 days. 567 of these 919 tissue samples were left to incubate up to 14 days. Wilkins Chalgren broth yielded 24.5% (139/567) positive cultures after 14 days of incubation. This was slightly more than the growth on blood agar after 14 days (22.9%--130/567) (p=n.s.) and significantly more than the growth in Wilkins Chalgren broth after 7 days of incubation (21.9%--124/567) (p<0.05). Based on these results, Wilkins Chalgren broth has been implemented as the SOP. Since the yield of positive cultures increased from 2 to 7 days of incubation in broth (1.8 times) and the variability of species cultured from 7 to 14 days of incubation shifted towards mostly microorganisms known to be common contaminants, we established the cut-off at 7 days of incubation in Wilkins Chalgren broth.     

Cell surface hydrophobicity and charge of Staphylococcus aureus and coagulase-negative staphylococci from bovine mastitis

The effects of seven growth media on cell surface hydrophobicity of a collection of Staphylococcus aureus and coagulase-negative staphylococci isolated from bovine mastitis were compared in the salt-aggregation test. Thirty-three per cent of Staph. aureus strains showed extremely high cell surface hydrophobicity (auto-aggregated) and 28% were moderately hydrophobic while 26% were hydrophilic after growth on horse blood agar at 37 degrees C for 18 h. There were great variations in the proportion and degree of the hydrophobicity depending on the medium used. Cultivations on/in capsule-inducing media caused a shift from a high to a low degree of hydrophobicity, although a microscopically detectable capsule or slime layer was seen in only one strain. This strain and encapsulated reference strains had a hydrophilic cell surface and migrated faster in free zone electrophoresis than cells of unencapsulated strains. Cells of strains grown on staphylococcus medium 110 agar migrated faster than those grown on horse blood agar regardless of their capsule production. Coagulase-negative staphylococci showed uniformly hydrophilic cell surface after cultivation on horse blood agar, but not when grown in tryptic soy broth or proteose peptone broth. It was concluded that most of the Staph. aureus strains from bovine mastitis under a variety of growth conditions in stationary phase culture constantly expressed hydrophobic cell surface.     

Agarose soy casein digest medium for replacement of blood agar for potency determinations of live Pasteurella vaccines.

Blood agar, prepared with Trypticase (BBL Microbiology Systems, Cockeysville, Md.) soy agar and 5% defibrinated bovine blood, is used for testing the potency of live Pasteurella multocida and Pasteurella haemolytica vaccines, but its potential for variation makes it undesirable to use in a standard assay method. Tests done with RPMI 1640 and Trypticase soy medium indicated that the benefits obtained by adding defibrinated blood to the Trypticase soy agar medium were more likely due to neutralization of toxic components than to the presence of transferrin or iron as growth factors. Reduction of toxic components in the Trypticase soy agar medium was accomplished by replacing agar with agarose and by autoclaving glucose as a separate solution to produce the replacement medium. The replacement medium was prepared by autoclaving three separate solutions--Trypticase soy broth without glucose; glucose; and agarose--cooling to 55 degrees C, and mixing and then pouring the mixtures into petri dishes. The growth obtained with this medium as judged by determination of the number of CFU and the colony sizes of P. multocida or P. haemolytica was equal to or better than those obtained with blood agar.