筛选试验在检测致病性气单胞菌中的应用

[目的]用分步筛选的方法检测致病性气单胞菌，以简化实验步骤，加快检测速度，减轻工作强度。[方法]初筛项目为氧化酶、无盐、6％NaCl胨水3试验，复筛项目为肌醇、精氨酸双水解酶、靛基质、赖氨酸脱羧酶等4项试验，凡氧化酶、靛基质阳性；无盐生长；6％NaCl胨水不生长；肌醇阴性；精氨酸双水解酶、赖氨酸脱羧酶不定的菌株为疑似气单胞菌，再进一步作系统生化鉴定试验，确定其种类。[结果]13种2024份海产品经初筛，检出疑似气单胞菌2668株，检出率为131．8％，其中542份样品同时检出2种以上疑似气单胞菌，进一步复筛筛去非气单胞菌880株，提高筛选率32．0％，系统生化试验后，确定致病性气单胞菌1667株，符合率为93．2％。[结论]分步筛选检测是一种较实用的过筛分离方法。与常法相比，简化了检测步骤，缩短了检测时间，减轻了工作量，节约了人力、物力，基本上解决了分离难的问题，取得了较满意的效果。     

从自来水中检出一株类志贺邻单胞菌

1990年9月,我们采用碱性胨水二次增菌和改良DC琼脂分离法,从局管内某单位采集的自来水水质监测样品中,检出一株类志贺邻单胞菌。 该分离菌株为革兰氏阴性短杆菌,在碱性胨水中均匀混浊生长,在改良DC琼脂上为光滑湿润、蓝色扁平菌落;在SS琼脂上为光滑湿润、无色透明菌落。主要生化特性为氧化酶试验阳性,O-F试验为F型,动力阳性,发酵葡萄糖产酸不产气;肌醇、靛基质、精氨酸双水解酶、鸟氨酸脱羧酶、赖氨酸脱羧酶、0％NaCl胨水、3％NaCl胨水为阳性;蔗糖、甘露糖、阿拉伯糖、VP、水扬素、七叶苷水解、枸椽酸盐、7％NaCl胨水、10%NaCl胨水     

鲜活小海产品中致病性弧菌和气单胞菌筛检方法的应用

目的：建立一套对致病性弧菌和气单胞菌同时筛选鉴定的检测方法。方法：将氧化酶阳性的可疑菌落先用革兰染色和OF试验初筛，再以0／129、无盐胨水和6％NaCl胨水试验复筛。结果：初筛剔除干扰菌67．57％（398／589）；复筛试验后，干扰菌的剔除率达到了88．79％（523／589），并能初步分出弧菌和气单胞菌。结论：该法能有效剔除干扰菌，简化步骤，且能对两种细菌实现同时筛选。     

从腹泻病人大便检出1株与肠出血性大肠菌O157交叉凝集的舒伯特气单胞菌

20 0 0年 7月 ,从就诊重症腹泻患儿稀便中检出 1株与肠出血性大肠菌 (ＥＨＥＣ)Ｏ157交叉凝集的舒伯特气单胞菌。将粪便按常规接种于ＳＳ、麦康凯琼脂和血琼脂平板 ,35℃ 2 4ｈ后 ,均生长出扁平、无色、中等大小菌落的优势菌 ,血琼脂平板呈溶血。挑取单个菌落染色为革兰氏阴性杆菌 ,动力活泼。菌株氧化酶、触酶阳性 ,葡萄糖产酸不产气 ,麦芽糖、明胶、甘露糖、Ｖ Ｐ、枸橼酸盐、硝酸盐还原、赖氨酸脱羧酶、精氨酸脱羧酶阳性 ,甘露醇、靛基质、尿素、七叶苷、鼠李糖、乳糖、蔗糖、纤维二糖、阿拉伯胶糖、水杨素、山梨醇、肌醇、鸟氨酸脱羧酶…     

甲鱼嗜水气单胞菌分离鉴定与药敏试验

从2只濒死甲鱼和1只刚死亡的甲鱼中分离出6株纯一优势细菌。该菌能在24h内对小白鼠致死,人工感染试验,症状与自然发病相似,结果表明分离菌为病原菌。该菌为G阴性杆菌,有动力,葡萄糖产酸产气,硝酸盐、赖氨酸脱羧酶、精氨酸双水解酶、七叶苷、水杨素、靛基质、V-P阳性,根据菌落形态、生化特性及API生化鉴定仪进一步鉴定,应为嗜水气单胞菌。药敏试验结果表明,该菌对氨基苷类、喹诺酮类及第三代头孢菌素类药物敏感。     

15株类志贺氏邻单胞菌的鉴定报告

近年来,我们从人和外环境中分离到15株类志贺氏邻单胞菌,其中来自腹泻病人的5株,来自食物中毒者的2株,来自健康查体者的3株,来自淡水的5株。 15株菌均为革兰氏阴性杆菌,两端钝圆,氧化酶、靛基质、动力、MR、硝酸盐还原、精氨酸、赖氨酸、鸟氨酸阳性,V—P、硫化氢、西蒙氏枸橼酸盐阴性,分解葡萄糖产酸不产气,分解肌醇,不分解甘露醇、阿拉伯胶糖、蔗糖、鼠李糖、棉子糖。4株迟缓发     

从腹泻病人和观赏动物分离出类志贺毗邻单胞菌

类志贺毗邻单胞菌是近年来新发现的一种重要肠道致病菌。本文从腹泻病人和观赏动物粪便中检出类志贺毗邻单胞菌。本菌为革兰氏阴性杆菌,一端有数根鞭毛,能运动。本菌的生化特性为氧化酶阳性,赖氨酸、鸟氨酸脱羧酶阳性,发酵肌醇。不发酵蔗糖、甘露糖、明胶和七叶苷,不能在TCBS和6%NaCl琼脂中生长。病人菌株和动物菌株的形态、培养及生化特征和耐药谱几乎完全相同。     

宁波市贝(甲)壳类海产品中致病性气单胞菌的分布

目的了解市售新鲜贝（甲）壳类海产品中致病性气单胞菌的携带及分布。方法用分步筛选的方法，筛检测贝（甲）壳类海产品中疑似致病性气单胞菌，再对疑似菌株进行系统生化定型。结果从13类2431份市售贝（甲）壳类海产品中检出致病性气单胞菌8种1946株，检出率为80．05％。检出的致病性气单胞菌以嗜水气单胞菌数量最多，占总数的38．26％；海产品带菌率最高为海瓜子，达145．31％。牡蛎、淡菜、海螺3类海产品检出的致病性气单胞菌种类和数量相对较少，同一样品中检出2种以上致病性气单胞菌有542份，占22．23％。结论检测发现市售新鲜贝（甲）壳类海产品携带致病性气单胞菌较普遍，贝壳类的致病性气单胞菌携带率高于甲壳类。结果提示，市售新鲜贝（甲）壳类海产品是一类诱发气单胞菌疾病的危险因子，而本地居民对该海产品的食用习惯多为生食或半生食，进一步增加了食用不安全性。因此，建议食用时应注意烹饪方法，以减少气单胞菌等疾病的发生。     

余杭市首次检出一株哈达尔沙门氏菌

患者,男,47岁。肠炎。大便黄色糊状,镜下白细胞少许,培养得一株哈达尔沙门氏菌(S.hadar)。 培养特性与形态染色:为革兰氏阴性小杆菌、无芽胞、无荚膜、有鞭毛,在肉汤中呈均匀混浊生长,在S.S琼脂上37℃24小时菌落直径2毫米,中心黑色,边缘光滑、整齐、透明,血琼脂上不溶血。 生化反应:氧化酶阴性,还原硝酸盐为亚硝酸盐,O—F试验为F型,分解葡萄糖产酸产气,甘露醇、山梨醇、肌醇、阿拉伯糖、鼠李糖、麦芽糖、木糖、甘露糖、蕈糖、纤维二糖、蜜二糖,赖氨酸脱羧酶、精氨酸脱羧酶、鸟氨酸脱羧酶、精氨酸双水解酶,甲基红、西蒙氏枸橼酸盐、粘液酸、醋酸钠、酒石酸、硫化氢阳性。葡萄糖酸盐、蔗糖、乳糖、棉子糖、ONPG、靛基质、卫矛醇、水杨苔、丙二酸盐、氰化钾、V—P、明胶、苯丙氨酸脱氨酶、侧金盏花醇、尿素酶阴性。 噬菌体(江西省卫生防疫站提供的肠杆菌科分     

用直接分离法检测贝(甲)壳类海产品中的致病性气单胞菌

[目的 ]了解贝 (甲 )壳类海产品中致病性气单胞菌的携带状况。 [方法 ]对市售贝 (甲 )壳类海产品标本用直接分离和 1%氯化钠胨水增菌后分离分别检测致病性气单胞菌。 [结果 ]从 40 0份样品中 ,直接分离法检出致病性气单胞菌 7种 480株 ,增菌分离法检出 6种 45 2株 ,检出率分别为 12 0 .0 %和 113.0 % ,两者符合率为 94.2 % ,差异无显著性 (P>0 .0 5 )。检出的致病性气单胞菌以嗜水气单胞菌为主 ,分别为 45 .4%和 46 .9%。 [结论 ]直接分离法能应用于贝 (甲 )类海产品中致病性气单胞菌的检测     

EAggEC生化特征及耐药性研究

目的 探讨EAggEC的生化特征及抗药性。方法 从门诊腹泻标本中分离EAggEC菌株，并作DNA探针和粘附试验、生化试验及药物敏感试验，对生化反应及药敏结果进行统计分析。结果 被检菌生化特征基本相似、氧化酶阴性、触酶阳性、IMVC反应为典型的 --，但有5％(2／40)菌株不分解山犁醇。而药敏试验情况表明，对乙酰螺旋霉素、麦迪霉素、林可霉素、多粘菌素、四环素等耐药，对环丙沙星、复达新、丁胺卡那敏感，可作为当前治疗的首选药。结论 本研究提示了5个监测点分离的EAggEC生化特征和耐药性基本情况。     

A comparison of ten USEPA approved total coliform/E. coli tests

Since 2002, the United States Environmental Protection Agency (USEPA) has approved ten enzyme-based total coliform and E. coli detection tests for examination of drinking water. These tests include: Colilert, Colilert-18, Colisure, m-Coli Blue 24, Readycult Coliforms 100, Chromocult, Coliscan, E * Colite, Colitag and MI Agar. The utility of the enzyme based test systems is based on both the ability of the test to detect the target organisms at low levels and the ability of the test system to suppress the growth of non-target organisms that might result in false positive results. Differences in the ability of some of these methods to detect total coliform and E. coli, as well as suppress Aeromonas spp., a common cause of "false positive" results, have been observed. As a result, this study was undertaken to elucidate the strengths and weaknesses of each method. Water samples were collected from three geographically and chemically diverse groundwaters in Wisconsin. One-hundred milliliter aliquots were individually spiked with both low concentrations (one to ten organisms) and high concentrations (fifty to one-hundred) of each of five different total coliform organisms (Serratia, Citrobacter, Enterobacter, E. coli, & Klebsiella). These spiked samples were used to test the capability of ten enzyme-based test systems to both detect and enumerate the spiked organisms. In addition, 100 ml samples were independently spiked with two different strains of Aeromonas spp. at six different levels, to assess the ability of each enzyme-based test to suppress Aeromonas spp. Analysis of the data indicated that wide variability exists among USEPA approved tests to detect and quantify total coliforms, as well as suppress Aeromonas spp.     

Faecal contamination indicators, Salmonella, Vibrio and Aeromonas in water used for the irrigation of agricultural products

The faecal contamination indicators (total coliforms, faecal coliforms, Escherichia coli, enterococci) and the genera Salmonella, Vibrio, Aeromonas were investigated in water samples used for irrigation. During 4 months, 52 samples were taken. The methods used were: multiple tube fermentation method for faecal contamination indicators and membrane filtration techniques for salmonella, aeromonas and vibrio. Two samples were positive for Salmonella spp., fourteen for Aeromonas spp. and no samples for Vibrio spp. No correlation was found between aeromonas and the indicators of faecal contamination. Regarding Aeromonas spp., 21.6% of the strains were adhesive and 12.6% cytotoxic: this confirms the possible role of aeromonas in human pathologies. These results are important to determine the quality of irrigation water in relation to human health. In fact, the spray or sprinkler irrigation produces bioaerosol, which can contaminate the crops that are likely to be eaten uncooked. In addition, the flood or furrow irrigation represents a risk to field workers.     

参龙健脑胶囊微生物限度检查的方法学验证研究

目的建立参龙健脑胶囊的微生物限度检验方法。方法按《中国药典》2005年版规定,分别采用常规法、离心加薄膜过滤法对样品进行微生物限度检查,根据5株阳性对照菌的试验组、菌液组、供试品对照组的结果进行其方法学验证试验研究。结果参龙健脑胶囊具有抑菌活性,离心加薄膜过滤法能有效地去除其抑菌活性。结论用该法进行微生物限度检查,可以客观地反映药物中微生物的污染状况,以达到检测目的。     

浦东新区腹泻样本中气单胞菌的分离鉴定及耐药性分析

目的:了解上海浦东新区感染性腹泻病人气单胞菌的感染及耐药情况,为相关防控策略的制定提供科学依据。方法:对2010年上海市浦东新区2个监测点,1245件急性腹泻样本进行气单胞菌检测,阳性菌进行药物敏感试验。结果:从1245件标本中检出气单胞菌93株,阳性率为7.47%,其中嗜水气单胞63株,温和气单胞26株,豚鼠气单胞4株。6月-8月份为高峰期。药敏试验结果:80%以上菌株对庆大,环丙沙星,阿米卡星、头孢噻肟敏感,对氨苄西林,复方新诺明、四环素、头孢噻吩、萘啶酸不同程度耐药。结论:气单胞菌感染存在季节性差异,与夏秋季腹泻关系密切。持续对气单胞菌及其耐药性的监测对细菌性腹泻病的预防与治疗具有指导意义。     

大肠杆菌O157胶体金免疫层析快速筛查方法的建立

目的建立一种大肠杆菌O157的胶体金免疫层析快速筛查方法。方法利用胶体金免疫层析技术,采用双抗体夹心法检测大肠杆菌O157,对该法进行敏感性、特异性和食品样品的适用性分析。结果该法能在15分钟内完成检测,检测不同的肠杆菌科细菌(非O157型大肠杆菌、沙门菌、志贺菌、变形杆菌、柠檬酸杆菌、肠杆菌、沙雷菌、耶尔森菌)、葡萄球菌、李斯特菌、气单胞菌、弧菌等共24种30株常见细菌,未发现有交叉反应,显示该法特异性良好。检测大肠杆菌O157的最低检出浓度为1×105cfu/ml。方法适用于奶粉、面粉、淀粉、咖啡粉、饼干、蛋糕、果冻、燕窝和果汁等食品样品的检测。结论新建立的大肠杆菌O157胶体金免疫层析试验简便、快速,特异性和灵敏度较好,适用于现场样品的快速筛查。     

Aeromonas detection and their toxins from drinking water from reservoirs and drinking fountains

Aeromonads are inhabitants of aquatic ecosystems and are described as being involved in intestinal disturbances and other infections. A total of 200 drinking water samples from domestic and public reservoirs and drinking fountains located in São Paulo (Brazil), were analyzed for the presence of Aeromonas. Samples were concentrated by membrane filtration and enriched in APW. ADA medium was used for Aeromonas isolation and colonies were confirmed by biochemical characterization. Strains isolated were tested for hemolysin and toxin production. Aeromonas was detected in 12 samples (6.0%). Aeromonas strains (96) were isolated and identified as: A. caviae (41.7%), A. hydrophila (15.7%), A.allosacharophila (10.4%), A. schubertii (1.0%) and Aeromonas spp. (31.2%).The results revealed that 70% of A. caviae, 66.7% of A. hydrophila, 80% of A. allosacharophila and 46.6% of Aeromonas spp. were hemolytic. The assay for checking production of toxins showed that 17.5% of A. caviae, 73.3% of A. hydrophila, 60% of A. allosacharophila, 100% of A. schubertii, and 33.3% of Aeromonas spp. were able to produce toxins. The results demonstrated the pathogenic potential of Aeromonas, indicating that the presence of this emerging pathogen in water systems is a public health concern.     

A comparison of ten USEPA approved total coliform/E. coli tests

Since 2002, the United States Environmental Protection Agency (USEPA) has approved ten enzyme-based total coliform and E. coli detection tests for examination of drinking water. These tests include: Colilert^®, Colilert-18^®, Colisure^®, m-Coli Blue 24^®, Readycult^® Coliforms 100, Chromocult^®, Coliscan^®, E*Colite^®, Colitag™ and MI Agar. The utility of the enzyme based test systems is based on both the ability of the test to detect the target organisms at low levels and the ability of the test system to suppress the growth of non-target organisms that might result in false positive results. Differences in the ability of some of these methods to detect total coliform and E. coli, as well as suppress Aeromonas spp., a common cause of “false positive” results, have been observed. As a result, this study was undertaken to elucidate the strengths and weaknesses of each method. Water samples were collected from three geographically and chemically diverse groundwaters in Wisconsin. One-hundred milliliter aliquots were individually spiked with both low concentrations (one to ten organisms) and high concentrations (fifty to one-hundred) of each of five different total coliform organisms (Serratia, Citrobacter, Enterobacter, E. coli, & Klebsiella). These spiked samples were used to test the capability of ten enzyme-based test systems to both detect and enumerate the spiked organisms. In addition, 100 ml samples were independently spiked with two different strains of Aeromonas spp. at six different levels, to assess the ability of each enzyme-based test to suppress Aeromonas spp. Analysis of the data indicated that wide variability exists among USEPA approved tests to detect and quantify total coliforms, as well as suppress Aeromonas spp.     

Aeromonas detection and their toxins from drinking water from reservoirs and drinking fountains

Aeromonads are inhabitants of aquatic ecosystems and are described as being involved in intestinal disturbances and other infections. A total of 200 drinking water samples from domestic and public reservoirs and drinking fountains located in S?o Paulo (Brazil), were analyzed for the presence of Aeromonas. Samples were concentrated by membrane filtration and enriched in APW. ADA medium was used for Aeromonas isolation and colonies were confirmed by biochemical characterization. Strains isolated were tested for hemolysin and toxin production. Aeromonas was detected in 12 samples (6.0%). Aeromonas strains (96) were isolated and identified as: A. caviae (41.7%), A. hydrophila (15.7%), A.allosacharophila (10.4%), A. schubertii (1.0%) and Aeromonas spp. (31.2%). The results revealed that 70% of A. caviae, 66.7% of A. hydrophila, 80% of A. allosacharophila and 46.6% of Aeromonas spp. were hemolytic. The assay for checking production of toxins showed that 17.5% of A. caviae, 73.3% of A. hydrophila, 60% of A. allosacharophila, 100% of A. schubertii, and 33.3% of Aeromonas spp. were able to produce toxins. The results demonstrated the pathogenic potential of Aeromonas, indicating that the presence of this emerging pathogen in water systems is a public health concern.     

中山地区人群HIV筛查与确证试验的比较

目的了解中山市普通人群和献血者人类免疫缺陷病毒(HIV)感染状况,提高实验室的检测水平。方法 2008～2012年期间,利用胶体金和ELISA技术筛查中山市普通人群和献血者HIV抗体,反应性标本用免疫印迹试验(WB)确证。结果共检测275 512例标本,初筛阳性率,普通人群0.11%(52/45 264),献血者0.12%(280/230 248);WB确证阳性符合率,普通人群为36.5%(19/52),献血者为12.5%(35/280);普通人群和献血者中HIV确证阳性率为0.04%(19/45 264)、0.02%(35/230 248);确证阳性者gp160、gp120、p24出现频率100%。结论中山普通人群和献血者感染趋势总体较为为平稳,普通人群HIV送检确证符合率较献血者高,HIV确证结果可以间接评价初筛试剂盒质量。