目的 基于环介导等温核酸扩增法(LAMP)建立副溶血性弧菌tlh基因的快速检测方法.方法 以副溶血性弧菌tlh基因为扩增的靶序列分别设计外引物、内引物和环引物各1对,通过引物特异性识别tlh基因上的6个独立区域在恒温条件下扩增靶序列,快速检测副溶血性弧菌.并对该法进行灵敏度、特异度试验,将该方法与荧光定量PCR法检出率进行统计学比较.结果 LAMP法最低检出限为10· fg/μl,灵敏度是荧光定量PCR的10倍;特异度实验中除检出2株副溶血性弧菌外,6株非副溶血性弧菌均未检出;LAMP法与荧光定量PCR法检出率差异无统计学意义(P＞0.05).结论 LAMP法具有高特异性、高效性、快速简便易检测等特点,适合现场快速检测,在食物中毒暴发处置中有深远的经济价值.

目的 联合应用荧光定量PCR/RT-PCR及脉冲场凝胶电泳(PFGE),对一起学校突发食物中毒事件进行病原检测和溯源分析,为突发中毒事件的应急处置提供参考.方法 采用现场流行病学调查方法对疫情经过进行调查,采集病例粪便/肛拭样本及剩余食品样本,进行细菌学培养、分离和鉴定.提取样本RNA,以荧光定量PCR/RT-PCR检测检测沙门菌、志贺菌、致泻大肠埃希氏菌、副溶血性弧菌、金黄色葡萄球菌诺如病毒核酸.对检出的副溶血性弧菌分离株进行血清学分型及PFGE分子分型分析,以BioNumerics软件进行同源性分析.结果 共采集43份样本,经常规培养鉴定和荧光定量PCR/RT-PCR分析,副溶血性弧菌阳性6份,分离株血清型均为O3:K6,除1株来自食品样本的菌株与其他菌株同源性为52.1％外,其余5株同源性为100.0％.结论 本起食物中毒事件是副溶血性弧菌污染导致.实时荧光PCR/RT-PCR方法结合PFGE分型技术可以快速、准确确定和溯源食物中毒的病原.

Objective To explore and found the quality control methods and affecting factors in isolation and identification of Staphylococcus aureus,so as to increase the isolation and identification accuracy of Staphylococcus aureus. Methods Food samples were selected from supermarkets,agricultural fairs and restaurants in Zhenjiang. Staphylococcus aureus was determined according to the standard GB / T 4789. 10—2010. National surveillance of food-borne pathogens Manual. Meanwhile,taking internal quality control in isolation and identification of Staphylococcus aureus,standard strains and reagents. All isolated strains should be submitted to Jiangsu CDC for reidentification as external quality control. Results From 2011 to 2013,a total of 37 Staphylococcus aureus were detected,including 32 coagulase test positive,5 coagulase test negative. External quality control was in line with the standard with rate of 100%( 37 /37). Conclusion National standard detection method for Staphylococcus aureus in food should be perfected,variety of detection methods should be taken to improve the detection accuracy of the strain identification results.

目的 了解2012年镇江市市售食品中食源性致病菌的污染情况.方法 从镇江市主要超市、农贸批发市场、餐饮单位(包括饭店、酒店及宾馆)采集食品样品共180份,按照GB 4789-2010《食品卫生微生物学检验标准》、国家食源性致病菌监测方案对样品进行副溶血性弧菌、金黄色葡萄球菌、蜡样芽孢杆菌、单核细胞增生李斯特菌、沙门菌检测.结果 检测的180份样品中,不合格率为28.89％(52/180),副溶血性弧菌、金黄色葡萄球菌、蜡样芽孢杆菌、单核细胞增生李斯特菌、沙门菌检出率分别为64.00％(32/50)、11.54％(15/130)、5.00％ (2/40)、2.86％ (4/140)、0.00％(0/140).结论 应加强食品的监测力度,确保市民的食品安全,防控食源性疾病的暴发.

Objective To analyze the molecular epidemiology traits of Vibrio parahaemolyticus in Zhenjiang and surrounding areas. Methods 40 Vibrio parahaemolyticus strains were collected from the samples of food poison incidents occurred in Zhenjiang and surrounding areas in 2011. Biochemical identification,tdh,trh and toxRS / new gene detection, serotyping and pulse-field gel electrophoresis( PFGE) were applied to these strains. Results The positive rate of tdh,trh, toxRS / new and orf8 were 85. 0%,5. 0%,50. 0% and 42. 5%,respectively. The prevalent serotype was O3∶ K6( 32 /40).The similarity of PFGE type between group A and B was more than 80%. Conclusion Strains from different regions in Zhenjiang and surrounding areas were mainly O3∶ K6 pandemic clone and highly homologous.

目的 了解镇江市食品中食源性致病菌的污染状况,为食源性疾病的监测提供科学依据.方法 依照国标和《江苏省食源性致病菌监测工作手册》,对采集样品进行病原菌分离鉴定.结果 180份食品样品,共检出致病菌50株,总检出率为27.78％;四大类食品中污染率最高的是生食动物性水产品,检出率为52.00％;熟肉制品、熟制米面制品的检出率依次为24.29％、17.50％;8种致病菌中,副溶血性弧菌检出率最高为52.00％,其次是金黄色葡萄球菌、蜡样芽胞杆菌、单核细胞增生李斯特氏菌,检出率分别为14.55％、7.50％、4.55％.结论 镇江市主要消费食品中生食动物性水产品、熟肉制品、熟制米面制品均存在不同程度的污染,应采取有效的控制措施,加强食品卫生监督管理,减少食源性疾患发生.

目的 观察低浓度含氯消毒剂对口腔综合治疗台水路消毒效果,为制定相关标准提供科学依据.方法 采用现场实际消毒效果检测方法,对三氯异氰尿酸消毒剂消毒口腔综合治疗台水路的效果进行了现场试验观察.结果 分别用有效氯含量为50、20和10 mg/L的三氯异氰尿酸泡腾片水溶液冲洗消毒以蒸馏水为水源的综合治疗台水路,消毒效果合格率依次为97.2％、91.7％和80.6％.用相同浓度的该消毒液冲洗消毒以市政供水为水源的综合治疗台水路,消毒效果合格率依次为94.4％、66.7％和61.1％.结论 用有效氯50 mg/L的含氯消毒剂水溶液冲洗消毒以蒸馏水或市政供水为水源的综合治疗台水路,均可达到消毒效果要求.

2011年11月1日,江苏省镇江市京口区某饭店发生群体性食物中毒,42人先后出现腹痛、腹泻等消化道不良反应,送至医院接受治疗,经流行病学调查和实验室检测,证实此次事件是由副溶血性弧菌引起的. 1 临床表现 42名患者均出现呕吐、腹痛、腹泻等消化道反应,其中3人有发热症状;大便为黄色水样便,每日4～8次.42人均进行了临床三大常规检查,结果显示尿常规、便常规均正常,仅血常规中白细胞和中性粒细胞增加.发病时间为3～8小时,病人经抗菌和输液治疗后,于次日全部痊愈.

目的 了解镇江市公共场所集中空调嗜肺军团菌的污染状况.方法 2012年采集市区7家公共场所集中空调的冷却塔水14份,并进行分离培养与鉴定.结果 14份样品中有5份检出军团菌,阳性率为35.7％,不同场所的阳性率依次为医院(66.7％)、商场超市(25.0％)、宾馆饭店(0).不同处理方法的检出率分别为酸处理(35.7％)、热处理(21.4％)、直接过滤(7.1％).检出5株嗜肺军团菌中,3株为LP7型,2株为LP1型.结论 镇江市医院集中空调内嗜肺军团菌污染最为严重,应加强管理,定期清洗空调系统.

A diverse collection of 261 Staphylococcus aureus strains from human, animal, food, and environmental sources were tested for the presence and type of SCCmec elements, antibiotic susceptibility to various antibiotics, and non-ß-lactam antibiotic resistance genes. About 18.39% (48/261) of strains were methicillin-resistant S. aureus (MRSA) including 29.75% (36/121) human strains of which 29 strains were hospital-acquired MRSA (HA-MRSA) and 7 strains were community-associated MRSA (CA-MRSA) and 19.67% (12/61) animal strains that all were CA-MRSA strains. The percentage of CA-MRSA strains from animals was significantly higher than that from human (p<0.01). Most of MRSA strains and a part of methicillin-susceptible S. aureus (MSSA) strains harbored unique combinations of non-ß-lactamase genes aac(6')/aph(2″), aph(3')-III, ant (4',4″), ermA, ermC, mrsA, tetM, and tetK. Antibiotic resistance genes were detected more frequently in HA-MRSA strains than in CA-MRSA strains (p<0.01). MRSA strains and MSSA strains had 22 and 39 antibiotic profiles to 15 tested antibiotics, respectively. The resistant proportion was higher in HA-MRSA strains than in CA-MSSA strains for various antibiotics, as well as higher in MRSA strains than in MSSA strains. Animal MRSA reservoirs (particularly pigs and cows) might represent an important source of human CA-MRSA. CA-MRSA strains might acquire more different resistance genes gradually, depending on the selective pressure of antibiotics in different regions or environments. CA-MRSA is not yet endemic in China, but could be prevalent in future, contributing to its acquiring more resistance genes and huge animal sources. Infection with multidrug-resistant MSSA strains acquired from food, animal, and human sources might also become a significant problem for human medicine, which warrants further study.

The levels of chemical pollutants in different type drinking water,which lead to rural cancers high incidence clusters distribution,were determined with analytical chemical methods and the trap(PT) and gas chromatography-mass spectrometry(GC-MS) instrument.Relationship of levels of chemical pollutants dose and cancer standardized mortality was tested with Spearman rank correlation.The results of the water quality tests showed that the CODMn,BOD5 and NH3-N levels in some of water samples from river water,pond water,and shallow well water exceeded the Chinese Class Ⅲ surface water standards.The microorganism indicators exceeded the drinking tap water criteria.NH3-N and NO3-N,NO2-N,which are the two types of precursors to synthetic NAD,and the NDMA and other direct carcinogen were tested.Thus it can conclude that river water,pond water and shallow well water in Z city rural can't server as drinking water or dinking water sources.The pollutions of different types of drinking water in wet season were higher than that in the dry season(P 0.05).The relationship between agestandardized cancers mortality and levels of drinking water pollutions dose in drinking water were positively correlated with grade(P 0.05).Total cancers mortality showed a dose-response trend with NO2-N,NDMA,or NDEA concentration in some villages.Direct drinking contamination surface water is one of the main reasons led to the high cancers incidence and clusters distribution of the City rural.

Objective To identify the cause of a food poisoning occurred in Zhenjiang district,and provide reference method for prevention of such mass incidents. Methods Biochemical identification,tdh and trh gene detection,serological typing and pulse-field gel electrophoresis( PFGE) were applied to the strains. Results All strains had virulent gene. The positive proportion of tdh and trh were 100. 0% and 7. 7% respectively. Strains isolated from patients all had tdh gene,only one had trh gene. Strains isolated from foods all had tdh gene,but not trh gene. The serological type of strains was O3: K6. Strains have high homology. Conclusion The PFGE technology can be effectively applied to tracing suspected sources of food contamination.

Objective:To study the distribution characteristics of systemic functional genes such as tdh,toxRS/new,orf8,HU-а,VP2905,MTase in O3:K6 pandemic clone,pathogenic and nonpathogenic strains of Vibrio parahaemolyticus.Methods:922 strains,including 180 O3:K6 strains,42 pathogenic strains and 700 nonpathogenic strains,were tested with PCR to analyze the distribution of these systemic functional genes based on our past studies.Results:Most of the O3:K6 pandemic clone strains had these systemic functional genes while only a few pathogenic and nonpathogenic strains harbored few of these genes.Conclusion:The systemic functional genes mainly existed in pandemic strains and played a great role in the forming course of pandemic clone and its infectious mechanism to human,while only individual genes existed in very few pathogenic and nonpathogenic strains occasionally.It could be confirmed as a pandemic strain when the strain had four or more than six systemic functional genes.

Objective To understand the molecular epidemiology traits of Vibrio parahaemolyticus in Jiangsu and surrounding areas.Methods Biochemical identification,tdh and trh gene detection,serological typing and pulse-field gel electrophoresis(PFGE) were applied to 63 Vibrio parahaemolyticus strains isolated from food poisoning incidents occurred in Jiangsu and surrounding areas in 2010.Results The positive rates of tdh and trh were 79.4%(50/63) and 1.6%(1/63) respectively.The main serological type of strains isolated from patients was O3∶K6(26/57),and that from food was O1∶KUT(4/6).The patterns of PFGE showed that the similarity of groups from A to E was more than 60%.Conclusion Strains isolated from different regions in Jiangsu and surrounding areas were in high homology.The food poisoning incidents occurred in these areas might be contaminated by Vibrio parahaemolyticus strains from the same source.

Objective:To understand the homology traits of Vibrio parahaemolyticus in Jiangsu.Methods:79 Vibrio parahaemolyticus strains were collected from the samples of food poison incidents in Jiangsu from 2009 to 2010.Biochemical identification,tdh and trh gene detection,serological typing and pulse-field gel electrophoresis(PFGE) were applied to the strains.Results:The positive rates of tdh and trh were 82.3% and 3.4% respectively.The main serological type of clinical strains was O3:K6,while that of food strains was O1:KUT.PFGE results showed that the similarity of group A to B was more than 60%.Conclusion:Strains of different regions and different time in Jiangsu have high homology,and food poison incidents occurred in this area may have the same pollution source.

Objective To understand the situation of pollutants in agricultural products from two regions in Zhenjiang with different cancer incidence,so as to provide evidence for reducing risk factors of cancer.Methods Lead,cadmium,mercury,aflatoxin and pesticides in agricultural food products were examined.Results Lead,cadmium,mercury were detected in 38,44 and 21 of 91 agricultural samples,respectively.There was no difference on the detection rate of metal pollutants between two regions.The amount of lead,mercury detected was not significantly different,but the amount of cadmium detected was different between two regions(P<0.01).Alatoxin B1 was detected in 2 samples,and B2,G1,G2 were not detected in 24 raw grain samples.Pesticide residues were detected in 11,30,3 and 3 from 24,48,12 and 7 raw grains,vegetables,fruits and tea samples,respectively.The pesticides detected were dimethoate,chlorpyrifos,methamidophos,dichlorvos,malathion,carbofuran,dicofol,chlorpyrifos-methyl and atrazine,etc.The detection rate of pesticide residues in agricultural food products was not different between the two regions,but the amount of pesticides detected was different between the two regions(P<0.05).The detection rate of pesticides in raw grains and vegetables were significantly different between the two regions(P<0.05).There was a positive correlation(r= 0.97) of organophosphorus pesticides with the cancer prevalence in the two regions,which showed a dose-effect relationship.Conclusion The contamination of cadmium and pesticide to agricultural food products might have some influence on the prevalence of cancer.

Objective To analyze the epidemiological features of handfoot-mouth disease(HFMD) in Zhenjiang from 2009 to 2010. Methods All the data of HFMD cases in Zhenjiang were analyzed by descriptive epidemiological method. Results The incidence rate of HFMD was highest in the period of April to June and in the age group of 1～3 years.Intriguingly,there was a slight predominance for boys and for children living in rural areas in HFMD infection. Conclusions The incidence of HFMD in Zhenjiang shows significant epidemiological features of region,season and population.It is important to take active prevention and control measures focused on the children in rural areas in the popular seasons.

<正>2009年,安徽、山西等多家医疗机构相继发生因血液透析液污染引发丙肝事件,为杜绝感染事件的发生,卫生部紧急下发了《血液净化标准操作规程(2010版)》[1]。为了解镇江市医疗机构血液透析现状,从2010年始,每季度对镇江市各级医疗机构透析液和透析用水进行检测,现将检测情况报告如下。

Objective:To phylogenetically analyze enterovirus 71(EV71) strains isolated from Zhenjiang in 2009 by amplifying vp1 gene.Methods:The throat specimens were selected from different patients of HFMD,and 11 nucleotide sequences of EV71 vp1 regions were successfully determined by RT-nested-PCR and sequencing.EV71 genotypes were analyzed using bioinformatic programs.Results:The sequence analysis showed that the 11 strains belong to C genotype,C4a subgenotype.Conclusion:The strains isolated from Zhenjiang are closely related to those isolated from Fuyang.All of them belong to C4a subgenotype and C4a subgenotype widely distributes and spreads in mainland.So it is important to enhance EV71 gene analysis and search after evolution track for EV71 in China.

Objective To establish a rapid,accurate method to identify the pathogen of hand,foot and mouth disease (HFMD) .Methods A multiplex real-time fluorescent quantitative PCR with TaqMan-LNA probe for identifying enterovirus 71 (EV71) and universal enterovirus (EV-U) simultaneously was developed.The standard curve was produced by RNA standard containing the target sequence to quantify the virus.The specificity,sensitivity and reproducibility of the developed RT-PCR method were estimated.Results The specificity of real time RT-PCR to detect CoxA16,CoxB2,EV71 and other human RNA virus were detectable by the developed method and the specificity was 100%,the sensitivity reached to 102 copies/μl.The repeat test for the standard of EV71 and EV-U showed that the coefficients of variation (CV) were 0.97%-2.02% for within-run and 0.89%-2.13% for between-run respectively.Conclusion The developed multiplex real-time PCR can be used for the diagnosis of HFMD because of its high specificity,sensitivity and reproducibility.