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1.
大肠杆菌O157:H7的疫病生态学研究进展及其控制对策   总被引:1,自引:0,他引:1  
大肠杆菌O157:H7是一种感染剂量低,致病性强,临床上无特效治疗药物的新型肠道致病菌,在世界范围内多次爆发流行,已构成严重的公共卫生问题。对大肠杆菌O157:H7的基本生物学特征,从非生物因素(包括传染源、气候等因素)和生物因素两个方面综述了影响大肠杆菌O157:H7传播的环境因子,最后从病原菌的监测诊断技术、废弃物堆肥处理、加强食品以及饮水安全管理等方面提出了控制大肠杆菌O157:H7的对策,最终为大肠杆菌O157:H7的疫病生态学研究提供了理论基础和参考依据。  相似文献   

2.
应用测试片快速检测食品中的大肠杆菌O157:H7   总被引:1,自引:1,他引:0  
目的应用大肠杆菌O157:H7测试片快速检测食品中的大肠杆菌O157:H7。方法对大肠杆菌O157:H7测试片的各项指标及影响因素进行测试,并将其应用于食品检测。结果大肠杆菌O157:H7测试片的检测灵敏度高,其对纯菌的检测低限可达3cfu/mL;特异性较强,与鼠伤寒沙门氏菌、志贺氏菌等21种非目的菌无交叉反应;快速,24h内可报告阴性检测结果。应用该测试片检测各种食品206份,检测结果与SN标准的符合率达到98.5%。结论应用测试片检测食品中的大肠杆菌O157:H7具有快速、方便、经济、无需昂贵设备等优点。该测试片可适应于食品中大肠杆菌O157:H7的快速初筛。  相似文献   

3.
为了研制一种快速、准确诊断大肠杆菌O157:H7的PCR方法,试验根据GenBank收录的大肠杆菌O157:H7全基因组序列,设计并合成了2对可扩增rfbE(O157)和Flic(H7)基因片段的特异性引物,建立双重PCR快速检测大肠杆菌O157:H7的方法,并研制出检测试剂盒。结果表明:该检测试剂盒对大肠杆菌O157:H7能特异性地扩增出rfbE和Flic基因的目的片段;对大肠杆菌O157:H30、O157:H9、O157:H25、O157:H19只能扩增出rfbE而不能扩增出Flic基因的目的片段;对大肠杆菌DH5α、猪链球菌2型和副猪嗜血杆菌没有扩增出任何目的片段;细菌DNA的最低检测极限是50 pg;在-20℃条件下保存1,3,6,9,12个月后,其敏感性都没有发生改变,均能检测到50 pg的大肠杆菌O157:H7DNA模板。  相似文献   

4.
通过将羧化聚苯乙烯乳胶与大肠杆菌O157:H7 Intimin蛋白单克隆抗体进行偶联,对最佳偶联抗体IgG量、最佳偶联时间等条件进行优化,建立了快速检测大肠杆菌O157:H7的乳胶凝集试验方法。结果表明,该方法具有快速、敏感、特异、简便等优点,适于临床应用,为大肠杆菌O157:H7的快速诊断提供了技术支撑。  相似文献   

5.
为了了解新疆不同牛场中大肠杆菌O157:H7的流行情况,对新疆乌鲁木齐、伊犁地区、塔城地区多个牛场采集的粪样及水样样本的检测,样品经EC肉汤增菌、免疫磁珠富集后,用SMAC平板和MUG培养基进行初步筛选,再用rfb E和fli C基因对筛选后的疑似大肠杆菌O157:H7菌株做PCR检测,同时结合生化试验的方法进行符合性检验。结果从新疆3个地区不同牛场的208份样品分离得到5株肠出血性大肠杆菌O157:H7,检出率为2.4%。其中粪样检出5株,检出率为2.4%;水样未检出。生化试验结果表明,5株分离出的大肠杆菌O157:H7菌株与国标规定的O157:H7特性相同。表明,部分被检牛场存在大肠杆菌O157:H7威胁,应该引起重视。  相似文献   

6.
根据GenBank公布的大肠杆菌O157:H7的Flic(H7)基因序列进行同源性比较分析,选择保守序列设计一对特异性扩增引物,通过优化反应条件,建立一个用于大肠杆菌O157:H7快速定量检测的实时定量PCR方法.该方法的最低检测极限是103CFU/mL,敏感性比常规PCR提高10倍.方法重复性好、特异性强,重复性检测的变异系数均小于2%;只能检测大肠杆菌O157:H7,对非大肠杆菌O157:H7血清型细菌、猪链球菌2型、副猪嗜血杆菌无反应.利用此方法对模拟样本进行定量检测,其结果与平板细菌计数基本一致,表明此方法可作为大肠杆菌O157:H7快速诊断和疫情监测的一种快速、准确、简便的检测工具.  相似文献   

7.
大肠杆菌O157:H7出血性肠炎是近年引起世界广泛关注的肠道传染病。自1983年美国Riley等首次报道大肠杆菌O157:H7出血性肠炎以来,相继在英国、加拿大、日本等国出现暴发性流行病例。大肠杆菌O157:H7是肠出血性大肠杆菌(EHEC)的一个主要血清型,牛、羊、  相似文献   

8.
五重PCR检测大肠杆菌O157:H7菌体抗原和毒素基因   总被引:4,自引:0,他引:4  
针对O157:H7大肠杆菌O抗原抗血清同多种细菌有交叉反应,而单一毒素基因并不是O157:H7所独有这一特点,建立了五重和四重PCRA-法,同时检测大肠杆菌O157:H7的O抗原、H抗原和4种毒素基因,可在较短的时间内检测、鉴定大肠杆菌O157:H7,并有较高的特异性,解决了传统细菌检测方法耗时长、灵敏度低,并有交叉反应等问题。  相似文献   

9.
以出血性大肠杆菌O157:H7对家兔进行试验感染,观察了大肠杆菌O157:H7对家兔的影响,通过玻片凝集试验分析了粪中大肠杆菌O157:H7的变化,结果表明,大肠杆菌O157:H7可导致兔的肠道反应和腹泻,在腹泻期排出的菌体最高,并可维护较长时间的带菌,说明家兔依然是志贺样毒素大肠杆菌的易感性动物、应引起高度重视。  相似文献   

10.
猪源大肠杆菌O157:H7广西分离株的鉴定   总被引:1,自引:0,他引:1  
为了解大肠杆菌O157:H7是否在猪场存在,采集有拉稀症状猪的粪便,接种到新生霉素mEC增菌肉汤,增菌培养后转接到O157:H7显色培养基上,对可疑菌株用因子血清和PCR方法做进一步鉴定,实验结果证实得到5株大肠杆菌为O157:H7血清型。动物试验发现,5个分离菌株对小白鼠的致死率为33.3%—100%之间。调查结果证实广西猪群中存在强致病性的O157:H7血清型大肠杆菌。  相似文献   

11.
The aims of the study were to determine the prevalence of enterohemorrhagic Escherichia coli O157:H7 (EHEC O157) and other Shiga toxin-producing E. coli (STEC) in feces of white veal calves in an operation in Ontario, to evaluate exposure of the calves to EHEC O157, and to investigate the milk replacer diet and antimicrobial resistance as factors that might influence the prevalence of EHEC O157. Feces from three cohorts of 20-21 calves were collected weekly for 20 weeks and processed for isolation of EHEC O157:H7 and detection of STEC by an ELISA. Exposure to EHEC O157 was also investigated by measuring IgG and IgM antibodies to the O157 lipopolysaccharide (O157 Ab) in sera by ELISA. The prevalences of EHEC O157 were 0.17% of 1151 fecal samples and 3.2% of 62 calves, and for STEC were 68% of 1005 fecal samples and 100% of 62 calves. Seroconversion to active IgG and IgM O157 Ab responses in some calves was not associated with isolation of EHEC O157. The milk replacer contained low levels of antibodies to EHEC antigens and without antimicrobial drugs, it did not inhibit the growth of EHEC O157 in vitro. Two E. coli O157:H7 that were isolated were totally drug sensitive whereas 60 commensal E. coli isolates that were examined were highly resistant. Antibodies in milk replacer that might be protective in vivo, and susceptibility to antimicrobial agents in the milk replacer may contribute to the low prevalence of EHEC O157 in white veal calves.  相似文献   

12.
A multiplex real-time PCR (R-PCR) assay was designed and evaluated on the ABI 7700 sequence detection system (TaqMan) to detect enterohemorrhagic Escherichia coli (EHEC) O157:H7 in pure cultures, feces, and tissues. Three sets of primers and fluorogenic probes were used for amplification and real-time detection of a 106-bp region of the eae gene encoding EHEC O157:H7-specific intimin, and 150-bp and 200-bp segments of genes stx1 and stx2 encoding Shiga toxins 1 and 2, respectively. Analysis of 67 bacterial strains demonstrated that the R-PCR assay successfully distinguished EHEC O157:H7 serotype from non-O157 serotypes and provided accurate profiling of genes encoding intimin and Shiga toxins. Bacterial strains lacking these genes were not detected with this assay. The detection range of the R-PCR assay for the three genes was linear over DNA concentrations corresponding from 10(3) to 10(8)CFU/ml of EHEC O157:H7. The R-PCR allowed construction of standard curves that facilitated quantification of EHEC O157:H7 in feces and intestinal tissues. Detection sensitivity of the R-PCR assay ranged from 10(4) to 10(8)CFU/g of feces or tissues without enrichment. Enrichment of feces in a non-selective broth for 4 and 16h resulted in the detection of levels (from 10(0) to 10(3)CFU/g of feces) considered sufficient for infection in humans. The R-PCR assay for eae(O157:H7), stx1, and stx2 proved to be a rapid test for detection of EHEC O157:H7 in complex biological matrices and could also potentially be used for quantification of EHEC O157:H7 in foods or fecal samples.  相似文献   

13.
Enterohaemorrhagic Escherichia coli (EHEC) infections in humans are an important public health concern and are commonly acquired via contact with ruminant faeces. Cattle are a key control point however cross-protective vaccines for the control of EHEC in the bovine reservoir do not yet exist. The EHEC serogroups that are predominantly associated with human infection in Europe and North America are O157 and O26. Intimin and EHEC factor for adherence (Efa-1) play important roles in intestinal colonisation of cattle by EHEC and are thus attractive candidates for the development of subunit vaccines. Immunisation of calves with the cell-binding domain of intimin subtypes beta or gamma via the intramuscular route induced antigen-specific serum IgG1 and, in some cases salivary IgA responses, but did not reduce the magnitude or duration of faecal excretion of EHEC O26:H- (Int(280)-beta) or EHEC O157:H7 (Int(280)-gamma) upon subsequent experimental challenge. Similarly, immunisation of calves via the intramuscular route with the truncated Efa-1 protein (Efa-1') from EHEC O157:H7 or a mixture of the amino-terminal and central thirds of the full-length protein (Efa-1-N and M) did not protect against intestinal colonisation by EHEC O157:H7 (Efa-1') or EHEC O26:H- (Efa-1-N and M) despite the induction of humoral immunity. A portion of the serum IgG1 elicited by the truncated recombinant antigens in calves was confirmed to recognise native protein exposed on the bacterial surface. Calves immunised with a mixture of Int(280)-gamma and Efa-1' or an EHEC O157:H7 bacterin via the intramuscular route then boosted via the intranasal route with the same antigens using cholera toxin B subunit as an adjuvant were also not protected against intestinal colonisation by EHEC O157:H7. These studies highlight the need for further studies to develop and test novel vaccines or treatments for control of this important foodborne pathogen.  相似文献   

14.
Enterohaemorrhagic Escherichia coli (EHEC) has been detected in both wild and domesticated rabbits in other countries. The aim of this study was to determine whether the most pathogenic E. coli serotype, O157:H7, occurs in the Dutch domesticated rabbit population and thus could form a public health risk. To this end, faecal samples were collected from rabbits from two rabbit farms and 741 rabbits of different breeds and origin and analysed for E. coli O157:H7, using a combination of enrichment, immunomagnetic separation, selective culture, and PCR. E. coli O157:H7 was not detected in any of the samples. The results indicate that Dutch domesticated rabbits probably do not play a role in the infection of humans with E. coli O157:H7.  相似文献   

15.
Over a 12 month period, 588 cattle faecal samples and 147 farm environmental samples from three dairy farms in southeast Queensland were examined for the presence of Shiga-toxigenic Escherichia coli (STEC). Samples were screened for Shiga toxin gene (stx) using PCR. Samples positive for stx were filtered onto hydrophobic grid membrane filters and STEC identified and isolated using colony hybridisation with a stx-specific DNA probe. Serotyping was performed to identify serogroups commonly associated with human infection or enterohaemorrhagic Escherichia coli (EHEC). Shiga-toxigenic Escherichia coli were isolated from 16.7% of cattle faecal samples and 4.1% of environmental samples. Of cattle STEC isolates, 10.2% serotyped as E. coli O26:H11 and 11.2% serotyped as E. coli O157:H7, and the E. coli O26:H11 and E. coli O157:H7 prevalences in the cattle samples were 1.7 and 1.9%, respectively. Prevalences for STEC and EHEC in dairy cattle faeces were similar to those derived in surveys within the northern and southern hemispheres. Calves at weaning were identified as the cattle group most likely to be shedding STEC, E. coli O26 or E. coli O157. In concurrence with previous studies, it appears that cattle, and in particular 1-14-week-old weanling calves, are the primary reservoir for STEC and EHEC on the dairy farm.  相似文献   

16.
Little is known of the association of enterohemorrhagic Escherichia coli O157:H7/NM (EHEC O157) with disease in naturally infected calves, although cattle have been known as a major source for EHEC O157 outbreaks in humans. In this study, we investigated the occurrence of EHEC O157 in calves associated with/without diarrhoea to examine if EHEC O157 is involved in calf diarrhoea and to characterize the isolates. Four hundred and ninety eight diarrhoeic and non-diarrhoeic young calves from 115 different farms were examined. Of 244 diarrhoeic calves, 24 (9.8%) were positive for EHEC O157, and of 254 non-diarrhoeic calves, 7 (2.8%) were positive. EHEC O157 was recovered from 12/76 (15.79%) of diarrhoeic calves less than 2-week-old, and no EHEC O157 was detected in this age group of non-diarrhoeic calves. This implicates EHEC O157 as a possible cause of the disease in naturally infected neonatal calves. The occurrence of EHEC O157 was relatively lower in the older calves (aged older than 8 weeks) and no significant difference was found in the occurrence rates between these diarrhoeic and non-diarrhoeic calves. PCR analysis of virulence markers revealed that the isolates carried various virulence genes such as Ehly, eae, stx1 and stx2, which underlines the potential importance of these attributes for the infection, colonization and possible pathogenesis of calf diarrhoea.  相似文献   

17.
Enterohaemorrhagic Escherichia coli (EHEC) infections in humans are frequently associated with direct or indirect contact with ruminant faeces and may result in haemorrhagic colitis and severe renal and neurological sequelae. Broadly cross-protective vaccines for control of EHEC do not yet exist and the molecular mechanisms that influence bacterial persistence in the intestines of ruminants are incompletely understood. We sought to determine the role in colonisation and protective efficacy of EspA, which forms a filamentous extension of the locus of enterocyte effacement-encoded type III secretion system that injects EHEC proteins into enterocytes. A non-polar deletion of espA severely impaired the ability of E. coli O157:H7 to colonise the intestines of calves. Vaccination of calves with highly purified recombinant EspA induced high-titre antigen-specific IgG1 (also reactive to native EspA) and salivary IgA responses, however these responses did not protect calves against intestinal colonisation by E. coli O157:H7 upon experimental infection.  相似文献   

18.
This study was aimed to understand the relationship of virulence gene distribution and genetic evolution between cattle originated Shiga toxin-producing Escherichia coli (STEC) and human originated enterohaemorrhagic Escherichia coli (EHEC) O157. This experiment collected 18 strains STEC in a dairy farm from Jiangsu province and 9 STEC reference strains (human, sheep, swine and avian), according to the method of U.S. Centers for Disease Prevention and Control Center (PulseNet), using the XbaⅠ enzyme digestion and pulsed field gel electrophoresis (PFGE) analysis, virulence genes were detected in some STEC isolates. The virulence gene distribution of O157 from different origin was remarkably different. The cattle originated STEC O157 and the human originated EHEC O157:H7 (EDL933W) had the most similar virulence gene distribution. In contrast, virulence genes were lack in cattle STEC O18 and O26, even though the cattle STEC O18 and O26 had the similar genotype as human EHEC O157:H7 (EDL933W). PFGE of Xba Ⅰ digested chromosomal DNA from 27 isolates of STEC exhibited 22 profiles. In general,the Dice coefficients of different originated STEC ranged from 72% to 100%.Cattle STEC O157 had a high similarity with two strains of human originated EHEC O157, while a low similarity was demonstrated between cattle STEC O157 and STEC O157 of swine and avian. The Dice coefficients of the cattle STEC O157 and the two strains of human EHEC O157 ranged from 83% to 95%. The Dice coefficients of cattle STEC O26 (Ⅶ,Ⅷ) and the two strains of human EHEC O157 were more than 82%. Therefore, it was concluded that the cattle STEC O157 and human EHEC O157 had a closer relationship in terms of virulence gene distribution and in genetic evolution.  相似文献   

19.
为了探讨牛源产志贺毒素大肠杆菌(Shiga toxin-producing Escherichia coli,STEC)分离株在毒力基因分布和遗传进化方面与人源EHEC O157菌株之间的关系,本试验选择收集来自江苏某奶牛场的STEC菌株18株以及人源、羊源、猪源、禽源STEC参考菌株9株,参照美国疾病预防控制中心PulseNet推荐的方法,运用XbaⅠ酶进行酶切并完成脉冲肠凝胶电泳(PFGE)分型和聚类分析;同时对部分STEC菌株进行毒力基因检测。结果表明,经毒力基因检测,不同来源的O157菌株毒力基因分布不尽相同,其中牛源STEC O157与参考株EHEC O157∶H7(EDL933W)的基因排谱最为相近;牛源STEC O18和O26的基因排谱与参考株EHEC O157∶H7(EDL933W)类似,但存在部分基因的缺失。对27株不同来源的STEC分离株进行PFGE,产生了22种不同的酶切图谱。总体来看,不同来源的STEC Dice相似性系数在72%~100%之间。牛源O157分离株与猪源及禽源O157菌株的相似度偏低,而与两株人源O157分离株的相似度偏高,Dice相似性系数在83%~95%之间,牛源O26(克隆群Ⅶ、Ⅷ)与人源O157的相似性系数 > 82%。显然,从牛群中分离到的部分STEC菌株与人源EHEC O157具有较近的遗传进化关系。  相似文献   

20.
Escherichia (E.) coli serotype O157:H7 is a globally distributed human enteropathogen and is comprised of microorganisms with closely related genotypes. The main reservoir for this group is bovine bowels, and infection mainly occurs after ingestion of contaminated water and food. Virulence genetic markers of 28 O157:H7 strains were investigated and multilocus enzyme electrophoresis (MLEE) was used to evaluate the clonal structure. O157:H7 strains from several countries were isolated from food, human and bovine feces. According to MLEE, O157:H7 strains clustered into two main clonal groups designated A and B. Subcluster A1 included 82% of the O157:H7 strains exhibiting identical MLEE pattern. Most enterohemorrhagic E. coli (EHEC) O157:H7 strains from Brazil and Argentina were in the same MLEE subgroup. Bovine and food strains carried virulence genes associated with EHEC pathogenicity in humans.  相似文献   

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