腹泻患儿粪便中沙门菌的血清型、药敏分析及脉冲场凝胶电泳分子分型研究

目的 研究2019年北京市朝阳区0-10岁腹泻患儿粪便中分离出的沙门菌的血清型、PFGE分子分型研究及耐药特点。方法 对分离自腹泻患儿病例的47株沙门菌进行血清分型,采用微量肉汤稀释法进行27种抗生素的药敏实验;采用PFGE脉冲场凝胶电泳进行指纹图谱分型研究。结果 47株沙门分为9种血清型,优势血清型两种,分别为肠炎沙门菌23株占48.94%,鼠伤寒沙门菌14株占29.79%。47株沙门菌对磺胺异恶唑耐药率(57.45%)最高,其次为氨苄西林(48.94%)和链霉素(48.94%)。23株肠炎沙门菌可分成8个PFGE指纹图谱,15株鼠伤寒沙门菌可分成14个PFGE指纹图谱。结论 北京市朝阳区0-10岁儿童食源性沙门菌血清主要为肠炎沙门菌和鼠伤寒沙门菌,各血清型的耐药性有所不同,PFGE指纹图谱呈多样性。     

一起肠炎沙门菌引起的校园食源性疾病暴发事件调查溯源分析

目的 调查一起肠炎沙门菌引起的学校食源性疾病暴发事件,并对危险因素开展分析溯源,为预防类似事件的发生提供科学依据。方法 运用描述性流行病学方法分析事件的流行病学特征;运用病例对照研究调查可疑食物;通过环境卫生学方法调查追溯食品污染的过程;对病例、食物、环境样品进行病原菌分离、血清分型和脉冲场凝胶电泳（PFGE）检测。结果 本起暴发事件共报告病例71例,流行曲线符合持续同源暴发模式的特点。病例对照、环境卫生调查和病因溯源研究提示学校超市售卖的三明治是引起本次事件的危险食品（OR=302.09,95%CI=75.18~1 213.97）。采集的病例、食品、环境样品中有8份（12.70%,8/63）检出了肠炎沙门菌,分离菌株的PFGE图谱条带完全一致,证实了学校超市出售的三明治使用的沙拉酱在制作过程中被肠炎沙门菌污染,是导致这次食源性疾病暴发的原因。结论 本次事件是由肠炎沙门菌污染食品引起的食源性疾病暴发,可疑食品为被自制沙拉酱污染的三明治。建议禁止各类未经充分热处理的含生鲜鸡蛋成分的食品进入学校,相关行政部门应加强对该类食品的卫生监督。     

某福利院一起肠炎沙门菌食源性疾病的调查分析

目的 对某福利院一起肠炎沙门菌食源性疾病进行调查和溯源,为研究相关食源性疾病提供参考。方法采用流行病学、食品卫生学和脉冲场凝胶电泳(PFGE)同源性分析等方法,分析本次食源性疾病事件。结果 确认本次事件中病例23名,患病率5.65%(23/407);现场采集病例肛拭子15份和厨房工作人员肛拭子3份、留样菜品3份、水果2份以及冰棒1份,其中从11份病例肛拭子中检出肠炎沙门菌,PFGE结果显示11株肠炎沙门菌的DNA条带图谱相似性为96.4%,聚类分析为同一型,结合流行病学调查,初步判断菌株来自同一克隆系。结论 综合流行病学、食品卫生学和实验室检测结果,确定为一起肠炎沙门菌引起的食源性疾病,福利院应加强对特殊人群的饮食安全管理,制定相应的食源性疾病突发事件应急处理预案,防止此类事故再发生。     

两起肠炎沙门菌所致食物中毒的病原学研究及溯源分析

目的探讨两起同地区连续发生的肠炎沙门菌食物中毒分离株之间的分子流行病学关系。方法参照GB 4789.4—2010《食品安全国家标准食品微生物学检验沙门氏菌检验》方法进行病原分离培养,对检出菌株进行表型鉴定;用VITEK-ⅡCompact全自动微生物分析系统检测菌株抗生素敏感性;采用脉冲场凝胶电泳(PFGE)技术对检出菌株进行分子分型和流行病学特征分析。结果共检出23株肠炎沙门菌,其中从25份患者肛拭样本中检出17株,8份从业人员肛拭样本中检出2株,8份留样食物中检出3株,厨具涂抹样本中检出1株。23株肠炎沙门菌血清抗原式均相同(9∶g,m∶-);生物学性状和药敏试验结果基本一致;PFGE图谱带型完全一致(相似度为100%),且与当地散发病例PFGE图谱带型相似。结论综合流行病学调查、病原学检测和分子分型结果,证实这两起食物中毒是由同一来源的肠炎沙门菌污染所引起。     

温州市食品中沙门菌污染状况及特征分析

目的了解温州市食品中沙门菌的污染状况,分析分离的沙门菌血清型分布、耐药性及脉冲场凝胶电泳(PFGE)分子分型特征。方法依据GB 4789.4—2016《食品安全国家标准食品微生物学检验沙门氏菌检验》进行菌株分离鉴定及血清学分型,采用微量肉汤稀释法进行药敏试验,PFGE法进行分子分型。结果 6类食品2 039份样品中,37份样品检出沙门菌,检出率为1.8%,其中生禽肉和生畜肉检出率较高,分别为6.9%(20/290)和3.4%(10/290)。37株沙门菌分属16种血清型,居前三位分别为鼠伤寒沙门菌、德尔卑沙门菌和肠炎沙门菌。81.1%(30/37)的菌株对17种抗生素产生不同程度的耐药,呈现24种耐药谱,多重耐药率为56.8%(21/37)。PFGE图谱分为31种PFGE带型,呈多态性。结论沙门菌在温州市食品中存在一定的污染率,耐药情况形式严峻,PFGE图谱的聚集性与沙门菌的血清型有一定的联系,与耐药谱之间的关联性并不明确。     

2017—2019年绍兴市沙门菌病人分离株的耐药性与分子分型分析

目的 了解绍兴市2017—2019年沙门菌病人分离株的分子分型特征及耐药情况。方法 收集245株分离自绍兴市食源性腹泻病例中的沙门菌,进行血清学分型,采用微量肉汤稀释法进行药敏检测,使用脉冲场凝胶电泳(PFGE)对分离株进行分子分型,利用BioNumerics V7.1软件进行聚类分析。结果 245株沙门菌可分为60种血清型,优势血清型为鼠伤寒沙门菌(32.24%)、肠炎沙门菌(10.20%)和伦敦沙门菌(6.94%)。225株分离株对25种抗生素存在不同程度的耐药,其中氨苄西林(77.14%)、四环素(73.88%)和链霉素(66.53%)的耐药率较高,且多重耐药率达76.33%。Xba Ⅰ酶切后的鼠伤寒沙门菌、肠炎沙门菌和伦敦沙门菌分别含48种、11种和12种不同PFGE指纹图谱。结论 绍兴市沙门菌血清型种类繁多,PFGE指纹图谱呈多样性,抗生素耐药呈现较集中且主要为氨苄西林-四环素-链霉素(AMP-TET-STR)。     

一起肠炎沙门菌引起的食物中毒调查分析

目的查明本次食物中毒事件的发生原因和可疑危险因素,同时对致病菌药物敏感性进行分析。方法通过发病时间和用餐时间结合病例临床特征分析查找可疑餐次,采集可疑食物和病例等标本进行实验室检测,食物及病例阳性标本做脉冲场凝胶电泳(PFGE)同源性分析,同时对致病菌做耐药分析。结果共确认81例病例,症状主要为发热≥38℃(100.00%,81/81)、腹痛(97.53%,79/81)、腹泻≥3次(100.00%,81/81)、呕吐(69.14%,56/81)等,食用被鸡蛋壳污染的蛋炒饭是发病的危险因素,厨师在制作蛋炒饭时,附有肠炎沙门菌的鸡蛋壳没有清洗干净,炒饭是将蛋液浇到米饭上,加热不彻底,最后导致肠炎沙门菌污染蛋炒饭。病例粪便检测为肠炎沙门菌阳性,和食物蛋炒饭中分离的肠炎沙门菌PFGE图谱完全一致,且与生鸡蛋壳涂抹样品中分离的肠炎沙门菌相似度达96%。头孢噻肟、环丙沙星、复方新诺明(TMP/SMZ)等对肠炎沙门菌具有较好的药物敏感性,萘啶酸耐药性强。结论食用被肠沙门菌污染的蛋炒饭是导致本次食物中毒的主要原因,建议加强对餐饮机构厨师的监督管理,规范卫生操作流程,提高卫生安全意识,防止类似事件再次发生。     

2016—2020年辽宁省沙门菌耐药性及分子分型研究

目的 分析辽宁省2016—2020年食源性疾病监测中沙门菌分离株的耐药情况及分子分型特征,为沙门菌引起感染性腹泻的防控、临床抗生素使用提供可靠科学依据。方法 对辽宁省2016—2020年临床腹泻病例分离的90株沙门菌进行血清学鉴定,应用脉冲场凝胶电泳（PFGE）进行分子分型,采用BioNumeries 7.6软件对菌株间的相似度进行聚类分析,采用微量肉汤稀释法进行药物敏感性检测。结果 90株沙门菌分为13种血清型,以肠炎沙门菌和鼠伤寒沙门菌为主。PFGE聚类分析得到54种带型,各带型间相似度为51.4%～100%,每种带型包含1～10株菌,同一血清型菌株的PFGE带型相似度较高,且存在多次聚集现象。药敏结果显示90株沙门菌呈现36种耐药谱,氨苄西林耐药率最高（66.7%,60/90）,其次为萘啶酸（62.2%,56/90）,头孢西丁全部敏感。多重耐药率达48.9%(44/90),其中鼠伤寒沙门菌和肠炎沙门菌多重耐药率分别为87.5%和45.8%。结论 辽宁省沙门菌引起腹泻病例呈散发态势,菌株多重耐药趋势明显,且耐药率较高,耐药谱广泛,应进一步加强分子溯源及耐药性监测。     

云南省肠炎沙门菌及德尔卑沙门菌脉冲场凝胶电泳分子分型及耐药性研究

目的 了解云南省沙门菌患者中分离最多的肠炎沙门菌及食品中分离最多的德尔卑沙门菌脉冲场凝胶电泳(pulse field gel electrophoresis, PFGE)分子分型及耐药状况。方法 参照中国细菌性传染病分子分型实验室监测网络Pulse Net China的沙门菌PFGE分子分型方法进行分子分型。分析耐药板最低抑菌浓度(minimal inhibitory concentration, MIC)值, 根据美国临床实验室标准化协会(Clinical and Laboratory Standard Institute, CLSI)的相应标准获得S、I、R结果。结果 56株肠炎沙门菌呈11种PFGE带型, 有4种优势带型, 对氨苄舒的耐药率最高, 为57.14%, 对亚胺培南最敏感。27株德尔卑沙门菌呈25种PFGE带型, 对复合磺胺的耐药率最高, 为64.29%, 对亚胺培南最敏感。结论 肠炎沙门菌分子分型有明显的优势带型, 本地区肠炎沙门菌对氨苄舒的耐药率最高。德尔卑沙门菌分子分型呈多样分布, 复方磺胺是本地区德尔脾沙门菌最耐药抗生素。2种沙门菌都对亚胺培南最敏感。     

2008—2018年四川省阿贡纳沙门菌分子分型及耐药分析

目的 了解2008—2018年四川省阿贡纳沙门菌(Salmonella agona)脉冲场凝胶电泳(PFGE)型别分布和耐药状况,为阿贡纳沙门菌引起的暴发预警、溯源调查及临床治疗提供依据。方法 对经过生化和血清学鉴定的61株阿贡纳沙门菌进行PFGE分子分型分析,采用微量肉汤稀释法测定14种抗生素对菌株的最小抑菌浓度。结果 61株阿贡纳沙门菌分离株共分为41个PFGE型,从不同年份、不同地区临床病例中分离到的部分菌株具有相同的PFGE型别,1株猪肉分离株与部分临床分离株具有相同的PFGE型别。28株阿贡纳沙门菌对14种抗生素均敏感,其余33株菌存在不同程度的耐药,其中四环素耐药率最高,为47.54%(29/61);多重耐药株共16株,占比为26.23%(16/61)。全部分离株均对亚胺培南敏感。结论 2008—2018年四川省阿贡纳沙门菌分离株PFGE型别呈多样性,在四川省为散发流行状态。耐药情况较严重,耐药性存在上升趋势。     

A pulsed field gel electrophoresis (PFGE) study that suggests a major world-wide clone of Salmonella enterica serovar Enteritidis

Since human infections by Salmonella enterica serovar Enteritidis (Salmonella Enteritidis) have been increasing world-wide over the past years and epidemiological studies have implicated the consumption of meat, poultry, eggs and egg products, elucidation of the predominant subtypes for this Salmonella spp. is important. In this study, 107 poultry and food isolates of Salmonella Enteritidis obtained from Germany were analyzed by pulsed field gel electrophoresis (PFGE), and the subtypes were compared with those of the 124 human isolates obtained in Taiwan. Results showed that for these 107 poultry and food isolates, when XbaI, SpeI and NotI were used for chromosomal DNA digestion followed by PFGE analysis, a total of 19, 20 and 19 PFGE patterns, respectively, were identified. Of them, 51 (47.7%), 52 (48.6%) and 42 (39.3%) strains belong to a single pattern of X3, S3 and N3, respectively, and 34 strains belong to a pattern combination of X3S3N3, which was the major subtype. When PFGE patterns of these 107 German isolates were compared with those of the 124 human isolates obtained in Taiwan, pattern combination of X3S3N3 was found as the most common pattern shared by isolates from both areas. PT4 is a major phage type for German and Taiwan isolates. Although most of the X3S3N3 strains are of this phage type, some strains of other PFGE patterns are also of this phage type. Since strains used in this study were unrelated, i.e., they were isolated from different origins in areas geographically far apart from each other, the PFGE study suggests a major world-wide clone of S. enterica serovar Enteritidis.     

苏州某市一起肠炎沙门氏菌食物中毒事件调查分析

目的用现场流行病学方法,分析本次食物中毒事件的原因和可疑危险因素,指导临床救治并提出预防措施。方法通过查阅医院就诊记录和医务室门诊记录、考勤记录、员工订餐记录等主动搜索病例,开展病例个案调查和现场卫生学调查,采集相关样品进行实验室检测等,判断引起本次食物中毒事件的可疑餐次和食物。结果通过访谈、个案调查、实验室检查等共搜索到可能病例67人(包括确诊病例27人),临床特征主要为腹泻(83.58%,56/67)、发热(79.10%,53/67)、腹痛(76.12%,51/67)等。可疑食品是3月3日晚餐的回锅肉,59份肛拭子分离到25份肠炎沙门氏菌,其中一株分离自厨师。经脉冲场电泳(pulsed-fieldgel electrophoresis, PFGE)分型, 25株肠炎沙门氏菌同源性为100%。经药敏分析显示, 25株肠炎沙门氏菌对头孢噻肟、头孢他啶具有较好的药物敏感性;对氨苄西林、氯霉素耐药性强。结论食用被肠炎沙门氏菌污染的回锅肉是导致本次事件的主要原因,建议监管部门加强对集体配餐单位的监督管理,提高从业人员食品安全意识,防范类似事件的发生。     

Antimicrobial resistance and phage and molecular typing of Salmonella strains isolated from food for human consumption in Spain

The aims of this study were to ascertain the population structure and antimicrobial susceptibility of Salmonella enterica serovars isolated in 2002 from food in 16 Spanish regions. Serovars were characterized by serotyping, phage typing, antimicrobial susceptibility, and pulsed-field gel electrophoresis (PFGE) typing, and 264 nonrelated strains were selected for further analysis. The main sources were eggs and their derivatives (21.6% of strains), poultry and related products (16.6%), and seafood (16.3%). High serotype diversity was detected (51 serotypes); the most common were Enteritidis (n = 96, 36.3%) and Typhimurium (n = 53, 20.1%), followed by a miscellaneous group of 49 different serotypes (n = 115, 43.5%). A 15% increase in Salmonella Enteritidis isolation was observed. Common phage types for Salmonella Enteritidis were PT1 (41.6% of isolates), PT4 (9.4%), PT6 (9.4%), and PT6a (9.4%), and common types for Salmonella Typhimurium were DTU302 (18.8%), DT104 (15.1%), and DT104B (13.2%). Salmonella Enteritidis strains were categorized into eight PFGE types with a similarity of 81 to 96%, and 73.9% of the strains were grouped into just one cluster. Salmonella Typhimurium isolates were divided into 13 PFGE types with a similarity of 64 to 86%, and one predominant clone contained 41.5% of the strains. Resistance rates for Salmonella Enteritidis, Salmonella Typhimurium, and the miscellaneous group were, respectively, 8.3, 69.8, and 13.9% for ampicillin, 3.1, 52.8, and 59% for streptomycin, 40.6, 22.6, and 10.4% for nalidixic acid, 15.6, 71.7, and 31.1% for tetracycline, 7.3, 18.8, and 9.5% for trimethoprim-sulfamethoxazole, 0, 50.9, and 4.3% for chloramphenicol, and 6.2, 71.7, and 17.4% for multiple (at least four) antimicrobials. All the strains remained susceptible to other beta-lactams and fluoroquinolones. Surveillance of S. enterica isolated from food is strongly recommended to reduce community exposure to antimicrobial resistant strains.     

一起纽波特沙门菌引起的食物中毒检测及菌株同源性分析

目的运用脉冲场凝胶电泳(PFGE)对一起食物中毒事件中分离的菌株进行同源性分析,为查明事件原因提供依据。方法采集患者、食品加工人员和剩余食物样本进行病原分离及鉴定,对分离到的菌株进行PFGE及耐药性分析。结果从采集的13份样本中检出4株纽波特沙门菌,其中1株来自病人样本,1株来自食品加工人员(厨师)样本,剩余牛肉、鸭肉各1株。4株菌株经PFGE分析,同源性为100%。4株分离菌株耐药谱相同。结论此次食物中毒由纽波特沙门菌引起,PFGE分型揭示菌株之间的流行病学联系,为事件的分析和溯源提供分子流行病学证据。     

Antimicrobial susceptibilities and epidemiological analysis of Salmonella enteritidis isolates in Korea by phage typing and pulsed-field gel electrophoresis

A total of 81 isolates of Salmonella Enteritidis were analyzed by antibiotic susceptibility, phage typing, and pulsed-field gel electrophoresis (PFGE). Thirty-two isolates came from broiler carcasses and pig feces, and 49 isolates were from humans in Seoul and suburbs of Seoul, Korea. Antibiotic resistance was most prevalent among human isolates. Of human isolates, 89.8% were resistant to more than two antibiotics, while 64.7% of poultry isolates and 13.3% of pig isolates showed multiple resistance to more than two antibiotics. The most common phage type (PT) was PT1, followed by PT30 or 33, PT21 and PT20a. The isolates showed six PFGE patterns with XbaI or SpeI digestion, and five PFGE patterns with NotI digestion. But a single pattern, PFGE X1, S1, or N1, was predominant and the rest of the PFGE patterns differed by only one or two bands. Results indicated the spread of a genetically related clone of Salmonella Enteritidis in foods and humans in Korea and that phage typing as well as PFGE may offer an improved level of discrimination for the epidemiological investigation of Salmonella Enteritidis.     

Iceberg lettuce as suggested source of a nationwide outbreak caused by two Salmonella serotypes, Newport and Reading, in Finland in 2008

A nationwide outbreak of Salmonella enterica serotypes Newport and Reading occurred between 17 October and 28 November 2008 in Finland. A total of 77 culture-confirmed Salmonella Newport and 30 Salmonella Reading cases, including one case with a double infection, were reported. All strains isolated from the patients were subtyped using serotyping, microbial resistance profiling, and pulsed-field gel electrophoresis (PFGE). Here, the PFGE patterns of the studied Salmonella Newport strains were identical, whereas four different PFGE profiles were found among the Salmonella Reading strains. Two elderly patients died within 2 weeks of the onset of symptoms. Three geographical clusters of cases with an epidemiological link were identified. The traceback investigation suggested that the factor connecting the cases was ready-chopped iceberg lettuce available for mass catering use. However, none of the tested food, environmental samples, or the samples taken from the staff of the processing plant contained Salmonella bacteria. Tracing back to outbreak sources with a short shelf life can be complex.     

2014—2019年成都市新都区鼠伤寒沙门菌耐药特性和分子分型研究

目的 了解新都区2014—2019年分离的54株鼠伤寒沙门菌耐药情况和分子分型特征.方法 对从病例监测、食物中毒和食品监测分离的54株鼠伤寒沙门菌用微量肉汤稀释法进行药敏试验,用脉冲场凝胶电泳(pulsed field gel electrophoresis,PFGE)进行分子分型.结果 54株鼠伤寒沙门菌耐药率为98...     

一起由B群沙门菌污染聚餐食品所致食物中毒调查分析

目的调查一起农村婚宴聚餐引起食物中毒的致病因子、致病食品及其污染来源,采取有效措施控制事件蔓延,预防今后类似事件的发生。方法开展现场流行病学调查,制定病例定义并主动搜索病例,采用描述流行病学方法分析本次事件病例的流行病学特征。通过个案调查访谈,对参加聚餐的108人开展病例对照研究,分析可能的致病食品。现场勘查和访谈厨师,了解婚宴菜品的制作过程、原料来源和所用水源等,并采集病例肛拭子、婚宴剩余食品、饮用水样进行实验室检测。结果本次食物中毒罹患率为21.6%(138/640),临床表现以腹泻、腹痛、发热为主。单因素分析结果显示94.4%(51/54)的病例和66.7%(36/54)的对照食用过由灌肚、里脊肉片、炸排骨、萝卜丝四种食品组成的凉菜拼盘,食用凉菜拼盘增加发病风险(比值比=8.50, 95%置信区间:2.32～31.02)。叉生分析结果显示同时食用拼盘中两种以上凉菜将增加发病风险(比值比=9.25, 95%置信区间:2.46～34.82)。4份病例肛拭子和1份凉菜拼盘中灌肚均检出B群沙门菌。5株检出的沙门菌脉冲场凝胶电泳(PFGE)指纹图谱为同一带型,提示病例和食品分离株在分子水平具有高度的同源性,为同一暴露源。结论本次事件是一起由B群沙门菌污染聚餐食品导致的食物中毒。凉菜拼盘是主要的致病食品,食品加工卫生环境与制作过程不规范操作是导致本次食物中毒发生的危险因素。当前农村自办宴席仍存在诸多食源性疾病发生的风险环节,应加强监管。     

ISOLATION AND CHARACTERIZATION OF NONTYPHOID SALMONELLA FROM HOSPITAL FOOD HANDLERS IN BEIJING, CHINA

Salmonella enterica isolates recovered from stool samples of healthy food handlers in two hospital dining halls in Beijing, China were characterized by serotyping, susceptibility to 15 antimicrobials and pulsed field gel electrophoresis (PFGE). Our data showed that 9.5% (29/305) of food handlers in these two dining halls were Salmonella carriers. Twenty-nine isolates were grouped into five serotypes including Agona, Derby, Enteritidis, Infantis and Senftenberg. All six Enteritidis isolates were resistant to chloramphenicol, nalidixic acid and tetracycline. Identical PFGE patterns were identified from food handlers in the same dining hall that indicated the possible transmission of Salmonella among food handlers and to customers. Our findings underscore the importance of food hygiene education and regular health examination among food handlers in dining halls in China.

PRACTICAL APPLICATIONS

Food handlers in dining halls should be considered as a high-risk group for Salmonella carrier state in China. Food hygiene education and regular health examination among food handlers in dining halls should be carried out.