电泳,凝胶,脉冲场

摘要： 目的 分析烟台市5起伤寒病原学特征及分子流行病学关联.方法 2018年自烟台市5例伤寒确诊病例样本和流行病调查样本分离获得6株伤寒沙门菌菌株,病例发病时间自2018年5月26日至7月24日,分别分布于烟台牟平区水道镇、烟台蓬莱区登州街道、烟台龙口东莱街道、烟台牟平区文化街道、烟台芝罘区福莱山街道.采用常规细菌分离方法和Xba I/Bln I双酶切脉冲场凝胶电泳(PFGE)技术对伤寒杆菌进行溯源分析,同时开展viaB毒力基因检测和27种临床常用抗生素敏感试验对伤寒杆菌进行病原学研究.结果 6株伤寒杆菌经PFGE-Xba I和PFGE-Bln I电泳分为4个PFGE模式,其中3株菌同源性100％;1株多耐药菌(外籍患者),1株单耐药菌(外省滞留史患者),1株完全敏感菌,同一PFGE模式的3株菌药敏表型一致,除对氨基糖甙类和喹诺酮类部分抗生素中介外,对其他抗生素均敏感;除外籍患者标本分离株viaB基因阴性外,其他菌viaB基因均阳性.结论 烟台寒杆菌对临床常用抗生素敏感性良好,仍需重视伤寒散发病例发生和伤寒输入感染.

摘要： 目的 探讨水产品中分离出的副溶血弧菌与腹泻患者分离出的副溶血弧菌的相关性.方法 通过血清学分型、聚合酶链反应检测、药敏试验和脉冲场凝胶电泳同源性分析348份水产品中分离出的副溶血弧菌与重庆市南岸区25例腹泻患者分离出的副溶血弧菌的相关性.结果 从348份水产品中分离鉴定出21株副溶血弧菌,检出率为6 .0%;从重庆市南岸区25例腹泻患者样本中分离鉴定出25株副溶血弧菌. 46株分离株以O4抗原血清型为主,占56 .5%(26/46) ;46株分离株均携带不耐热溶血毒素毒力基因,腹泻患者的分离株还携带相对耐热直接溶血毒素(TDH )或相对耐热直接溶血毒素毒力基因;46株分离株对头孢唑啉全部耐药,对氨苄西林部分耐药或中度敏感;腹泻患者同源性较高,水产品带型多、相似度低,亲缘关系不紧密.结论 重庆市南岸区水产品和食品监测样品的副溶血弧菌污染率不高,水产品中副溶血弧菌分离株致病力较弱,腹泻患者的分离株致病力较强,血清型、耐药性和同源性无明显相关性,只有突发公共卫生事件同批次腹泻患者的分离株同源性存在极高的相关性.

摘要： 针对一起于2016年6-9月发生在江阴市新桥镇的伤寒疫情开展暴发确认和传播因素分析.首先收集分离自确诊病例的14株伤寒沙门菌;再采集与疫情相关的外环境样本65份和食品样本13份,运用实时荧光PCR法检测样本中伤寒沙门菌特异基因,采用常规分离培养法分离伤寒沙门菌.对所有伤寒沙门菌分离株进行药敏试验及脉冲场凝胶电泳(PFGE)分析.从外环境样本(K熟食店加工厂井水)中分离得到1株伤寒沙门菌.15株伤寒沙门菌均对萘啶酸耐药,经脉冲场凝胶电泳后共获得2种带型,其中13株病例分离株和环境分离株的PFGE指纹图谱完全一致,另外l株病例分离株的PFGE带型存在1个条带的差异.结合流行病学调查和实验室检测结果,判定本次暴发疫情是由同一伤寒沙门菌遗传克隆的传播而导致,食品加工厂应属于是传播环节之一.

摘要： 目的 评价基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)技术在碳青霉烯耐药肺炎克雷伯菌(CRKp)同源性分析中应用的可行性.方法 回顾性分析.收集2017年1月至12月全军细菌耐药监测网上海长海医院临床标本分离的肺炎克雷伯菌非重复菌株57株,新疆乌鲁木齐市第一医院临床标本分离的肺炎克雷伯菌非重复菌株36株;纳入标准:对亚胺培南、美罗培南或厄他培南当中任一药物耐药菌株并鉴定质谱蛋白峰数大于100的菌株,共计39株;以脉冲场凝胶电泳(PFGE)结果为依据,将待测菌株分为以下三组:高度同源组(包括产OXA-48型CRKp菌株17株)、中度同源组(产KPC型CRKp菌株16株)和非同源性组(包括携带其他耐药机制的CRKp菌株8株).所有待测菌株经MALDI-TOF MS分析后,所获结果分别采用BioNumeric软件和质谱仪自带的同源性分析软件(SARAMIS数据库软件)的两种分析方法进行同源性分析.以PFGE分析结果为参考标准,评价其他三种分析方法所获结果的可靠性.结果 对高度同源组菌株,采用SARAMIS两种分析方法分析KA-A型质谱数据Identical mass值高于80％,相似度高于85％,能够获得与PFGE分析结果较一致的结果;对非同源组的菌株,SARAMIS两种分析方法分析质谱数据Identical mass值低于80％,相似度不足85％,所获分型结果与PFGE结果有较好的一致性,能够反映菌株间的差异性;对中度同源组菌株,BioNumeric软件和SARAMIS数据库软件分析质谱数据KC-A型的identity mass值不足70％;相似度不足90％,所获得的结果与PFGE结果之间均存在着较大的差异,无法获得与PFGE结果相一致的分析结果.结论 MALDI-TOF MS结果采用BioNumeric软件和SARAMIS数据库软件进行同源性分析可作为具有高度同源性和非同源菌株间同源关系的初筛方法,但仍需要其他方法加以验证;对同源关系多样的菌株,上述方法仍缺乏稳定性,不适于进行菌株间同源关系分析.%Objective To evaluate the ability of Matrix-assisted laser desorption ionization-time of flight mass spectrum (MALDI-TOF MS) on homology analysis of carbapenem-resistant Klebsiella pneumoniae(Carbapenemase-Resistant K. pneumoniae, CRKp). Methods During January 2017 to December 2017, a total of 57 strains isolatedfromChanghai Hospital, Shanghai and 36 strainsfrom Urumqi First Hospital, Xinjiang were included for retrospective study. Inclusion criteria:strains which resistant to one of the carbapenems and with more than 100 peaks in identification mass spectrum. A total of 39 isolates were selected and divided into three groups according to the PFGE typing results:(1) Highly homologous group (17 strains with blaOXA-48 gene); (2) Moderate homology group (16 strains with blaKPC gene); (3)The non-homology group (8 strains with other resistance mechanisms). Mass spectrometry results were analyzed using BioNumeric and the two classification methods in SARAMIS, respectively.The PFGE analysis results were regarded as standard when evaluate the reliability of the other three analytical methods. Results Forhighly homologous group, the analysisresults of KA-A type dientical mass by two classification methods in SARAMIS were accord with PFGE,and the identity mass value is>80％, the similarity is>85％. However, for non-homologous group, the identity mass value is<80％, the similarity is<85％ when compared with PFGE, which indicated that the method based on MALDI-TOF MS was not premium for this group of strains. For moderate homologous strains,the results of two SARAMIS methods for KC-A type havepoor consistency with PFGE, for the identity mass value is<70％, the similarity is<90％. Conclusions Based on MALDI-TOF MS using BioNumeric software and SARAMIS database software could be regarded as a screening method for phylogenetic analysis among CRKp strains with high homology. While, it should not be applied to evaluate homology of the strains with genetic diversity for lacking stability.

摘要： 目的 分析山东省动物粪便中非O157产志贺毒素大肠埃希菌(STEC)的菌株特征及耐药状况.方法 于2015—2016年,采用方便抽样方法在山东省微山县、莱州市共采集1022份新鲜动物粪便标本,共分离到24株非O157 STEC.通过血清凝集实验进行血清分型,采用微量肉汤稀释法进行药物敏感性试验,后采用双纸片协同试验进行超广谱β-内酰胺酶(ESBLs)检测.采用PCR方法检测耐药基因.采用PFGE进行分子分型并采用多位点序列分型(MLST)确定等位基因谱及菌株序列型.使用CLC Sequence Viewer及Conting Express生物信息软件分析不同ST序列群及菌株间的进化关系.结果 24株非O157型STEC菌株中23株来自猪粪,1株来自牛粪;毒力基因均为stx 2型;对磺胺异(恶)唑耐药的菌株较多,为22株,对复方磺胺异(恶)唑和萘啶酸耐药的菌株均为18株,对氯霉素耐药菌株为13株,对四环素耐药菌株为19株.存在多重耐药现象,耐药谱为:磺胺异(恶)唑-四环素-复方磺胺异(恶)唑-萘啶酸-氯霉素,对头孢吡肟和亚胺培南均为敏感.ESBLs确证实验表明,有4株非O157型STEC产ESBLs.PCR检测7种耐药基因,4种四环素类耐药基因(tetA、tetB、tetC、tetD)均有检出, ESBLs类耐药基因(blaSHV-1、bla CTX-M、bla TEM)均为阴性.24株菌分成15个PFGE型别,其聚类结果比较分散,无优势PFGE型别.MLST型别有11种,其中ST540、ST5133型别的菌株最多,各为4株菌,聚类为1个MLST基因组.结论 山东省部分地区在动物粪便中检出的非O157 STEC血清型比较分散,对常见的抗生素耐药率比较高,具有明显的分子多态性,应重点加强对这类菌株的监测和管理.%Objective To understand the infection status,characteristics and drug resistance of non-O157 Shiga toxin-producing E. coli (STEC) in animal feces in Shandong Province. Methods From 2015 to 2016,convient sampling method was used to collect 1 022 fresh feces of animals in Weishan county and Laizhou city, and 24 non-O157 STEC were isolated. The serotypes of non-O157 STEC strains were confirmed through serum agglutination test. The susceptibility was explored through the antimicrobial sensitivity experiments. ESBLs activity was confirmed by double-disc diffusion. PCR method was used to detect the resistance genes.PFGE typing was operated to assess the relatedness and variability of the strains. The multi-locus sequence typing (MLST) was adopted to get the allelic profile and ST sequence of strains. Analysis was made on the evolutionary relationship between different ST groups was made through CLC Sequence Viewer and Counting Express.Results A total of 24 non-O157 STEC were isolated from animal feces. 23 strains were from pig feces, and 1 strain was from cow feces, and the serotypes were more dispersed. All of the 24 strains carried s tx 2 genes. The highest resistance rate was sulfamethoxazole(22 strains),the mount of cotrimoxazole and nalidixic acid was 18 strains,chloramphenicol was 13 strains, tetracycline was 19,and there was a phenomenon of multiple drug resistance.The drug resistance spectrum was sulfamethoxazole tetracycline-compound novammin-naphthidine-chloramphenicol. All strains were sensitive to cefepime and imipenem.The ESBLs confirmatory test showed that 4 strains of non O157 STEC produced beta lactamase.PCR detected 7 resistance genes,and 4 tetracycline resistance genes(Tet A,Tet B, tetC and tetD)were detected.The beta lactamase resistance genes(blaSHV-1,bla CTX-M,bla TEM)were all negative.24 strains were divided into 15 PFGE types,and their clustering results were more dispersed and no dominant PFGE type. There were 11 kinds of MLST types, most of them are ST540 and ST5133 types, each of which was 4 strains,and clustered into 1 MLST genomes.Conclusion The serotypes of non-O157 STEC in animal feces O157 STEC were dispersed, and the resistant rate to common antibiotic was high. MLST typing results presents obvious polymorphism.Surveillance and manage ment of these strains should be strengthened.

摘要： 幽门螺杆菌(Hp)可以在人群中广泛传播,并会引发一系列的胃肠道疾病甚至胃癌.不同基因型的Hp感染可能会引发不同类型的疾病,而基因分型技术是研究这类问题的重要方法.本文主要介绍了多位点序列分型、脉冲场凝胶电泳、随机扩增多态DNA、扩增片段长度多态性和全基因组测序这五种基因分型技术,通过综述这几种技术在Hp基因分型中的应用进展,比较它们之间的优缺点,为今后研究Hp的致病机制、传播机制以及流行病学调查提供方法学依据.%Helicobacter pylori ( Hp ) is widely disseminated in human, and Hp infection causes various gastrointestinal diseases, including gastric cancer. Different genotypes of Hp may cause different diseases, so the genotyping is important for clinical and basic research of Hp. This article introduces the methods for Hp genotyping, including multilocus sequence typing, pulsed-field gel electrophoresis, random amplified polymorphic DNA, amplified fragment length polymorphism, and whole-genome sequencing. By reviewing the application of these techniques in Hp genotyping and comparing their advantages and disadvantages, the article provides a theoretical basis for research into the pathogenesis, antibiotic resistance, and epidemiology of Hp infection.

摘要： Objective To investigate the antimicrobial resistance and pulsed field gel electrophoresis (PFGE) patterns of S.paratyphi A strains in Zhengzhou city isolated from sentinel hospitals in 2013-2015. Methods According to Salmonella molecular typing and K-B drug susceptibility testing method published by international PulseNet bacterial infectious disease monitoring network and USA Clinical and Laboratory Standards Institute (CLSI2015), we analyzed drug sensitivity and PFGE molecular characteristics of 67 S.paratyphi A strains(11 strains in 2013, 7 strains in 2014, 49 strains in 2015) isolated from blood and stool samples in two sentinel hospitals of fever with rash syndrome surveillance system established in Zhengzhou city in 2013-2015. Results The results showed 67 strains of S.paratyphi A had different levels of resistance to 13 kinds of antibiotics, 65 strains were multi-drug resistant strains (97.0%), 5 isolates were resistant to 2-3 kinds of antibiotics (7.5%), 41 isolates were resistant to 5-8 kinds of antibiotics (61.2%),11 isolates were resistant to 9-10 kinds of antibiotics(16.4%),8 isolates were resistant to 11-12 kinds of antibiotics(11.9%). 67 strains of S.paratyphi A were divided into 10 molecular patterns (PTYA1-PTYA10) by digestion with XbaⅠ restriction endonuclease and pulsed field gel electrophoresis, each pattern contains 1-48 strains with similarity ranged from 94.31%-100%. PTYA3 contained 48 strains, which was predominant band type; PTYA1, 9 contained 6 strains; PTYA 2, 4, 5, 6, 7, 8, 10 contained 1 strains among them. Conclusion The status of drug resistance of clinical isolates of S.paratyphi A in Zhengzhou city was rather serious, PFGE patterns showed diversity and dominant characteristics. The PFGE patterns of partial strains and its corresponding anti-drug spectrum have certain relevance and cluster relationship.%目的 分析2013—2015年郑州甲型副伤寒沙门菌临床分离株的耐药状况与脉冲场凝胶电泳(PFGE)分子型别特征.方法 于2013—2015年,从郑州发热伴出疹症候群监测系统2个哨点医院内,分离经病例血培养和粪便培养的67株甲型副伤寒沙门菌,其中2013年11株,2014年7株,2015年49株.根据国际PulseNet网络实验室公布的沙门菌PFGE分型操作规程与美国临床实验室标准化协会沙门菌K-B法药敏试验方案,对67株甲型副伤寒沙门菌进行药敏试验与PFGE脉冲场凝胶电泳分子分型分析.结果 67株甲型副伤寒沙门菌对13种抗生素均有不同程度的耐药,有65株为多重耐药株(97.0%),其中耐2~3种的为5株(7.5%),耐5~8种的为41株(61.2%),耐9~10种的为11株(16.4%),耐11~12种的为8株(11.9%).经XbaⅠ酶切与脉冲场凝胶电泳后,67株甲型副伤寒沙门菌共获得10种带型(PTYA1~10),每种带型包含1~48株菌株,相似度为94.31%~100%.其中PTYA3包含48株菌,为其优势带型;PTYA1、9包含6株菌;PTYA2、4、5、6、7、8、10各包含1株菌.结论 郑州市临床分离的甲型副伤寒沙门菌耐药状况普遍比较严重,PFGE带型呈现多样性的同时又具有较显著的优势带型特点,部分带型与其对应的耐药谱具有一定的关联性与聚集性.

摘要： 目的 分析沈阳地区临床与环境分离的嗜肺军团菌(LP)的同源性.方法 收集2012年7月-2014年9月来自沈阳市2例LP感染患者、2家医院及13家公司生活用水和空调冷凝塔水中分离得到的22株LP.对菌株进行复苏并应用Asc Ⅰ内切酶进行脉冲场凝胶电泳,采用SPSS 19.0软件对指纹图谱进行聚类分析.结果 22株菌中20株为LP1型,其余2株分别为LP7型和LP8型;共分为14种基因型,其中SYC型有7株,同一血清型菌株可呈不同基因型;同一地点可分离出不同血清型和基因型的菌株;同一医院冷却塔和生活用水中分离的LP菌株有同源性.结论 沈阳地区从临床及环境分离的LP具有基因多态性;同一医院不同水系中存在的LP菌株具有同源性.%Objective To study the homology of Legionella pneumophila (LP) strains which were isolated from clinical specimens and environmental water.Methods Twenty-two Legionella strains were collected in which 2 strains were isolated from 2 cases of LP infection patients,and others were isolated from domestic water or air conditioning cooling tower water of 2 hospitals and 13 companies from July 2012 to September 2014 in Shenyang.They were recovered and analyzed by the pulse field gel electrophoresis (PFGE) with Asc Ⅰ restriction enzymes.Data were cluster analyzed using SPSS 19.0.Results Twenty of 22 strains were LP1,and others were LP7 and LP8.Twenty-two strains could be divided into 14 genotypes,and 7 strains belonged to SYC banding pattern.The same bacterial serotype presented with different genotypes.Different bacterial serotypes and genotypes of LP strains can be found in the same place,and homology could be observed between LP strains isolated from cooling tower water and domestic water within the same hospital.Conclusion In Shenyang area,LP strains which are isolated from clinical cases and environmental water have the characteristics of gene polymorphism.Legionella strains isolated from different water system in the same hospital have homology.

摘要： Objective To investigate the antimicrobial resistance status and pulsed field gel electrophoresis (PFGE) patterns of Salmonella Enteritidis (S.Enteritidis) strains in Henan province. Methods S. Enteritidis strains were isolated from seven sentinel hospitals from March 2011 to December 2013. According to molecular typing and Salmonella (Kirby-Bauer, K-B) drug susceptibility testing method published by the international PulseNet bacterial infectious disease monitoring network and USA Clinical and Laboratory Standards Institute (CLSI), we analyzed drug sensitivity of 8 kinds antibiotics and PFGE molecule characteristics of 120 S. Enteritidis isolates from seven sentinel hospitals. Results Among 120 strains of S. Enteritidis, 77 were isolated from male patients, 43 from female patients. A total of 78 strains S. Enteritidis were isolated from young children ranged from 0 to 5 years old (65.0%), including 57 strains isolated from 6 months to 2 years old (47.5%). The isolated time mainly centralized on May to October of the year, 11 strains isolated in March-April (9.2%), 48 were in May-July (40.0%),54 in August-October (45.0%), 7 in other months, with a typical summer seasonal characteristics. The resistance rate of 120 strains S. Enteritidis to ampicillin was 50.0% (n=60); to ceftazidime was 14.2% (n=17), to cefotaxime was 18.3%(n=22);to cefepime was 5.8%(n=7);to nalidixic acid was 61.7%(n=74);to ciprofloxacin was 8.3%(n=10), to norfloxacin was 5.8% (n=7); to gentamicin was 42.5% (n=51); to streptomycin was 21.7% (n=26); to chloramphenicol was 30.0% (n=36); resistance to methicillin benzyl ammonium was 11.7% (n=14), compound sulfamethoxazole resistance rate was 71.7%(n=86);the tetracycline resistant rate was 47.5%(n=57). All 120 strains of S. Enteritidis had different levels of resistance to 8 kinds of antibiotics, all strains were multidrug resistant strains, 28 isolates were resistant to 3-4 kinds of antibiotics (23.3%), 38 isolates were resistant to 5-6 kinds of antibiotics (31.7%), 39 isolates were resistant to 7-8 kinds of antibiotics (32.5%). All 120 strains of S. Enteritidis were divided into 44 molecular patterns by digestion with XbaI and pulsed field gel electrophoresis. each pattern contained 1-35 strains with similarity ranged from 54.3%-100%. EN14 and EN19 were the main PFGE types, including 35 and 29 strains respectively. Conclusion The status of drug resistance of clinical isolates of Salmonella in Henan province was rather serious, PFGE patterns showed advantages and partial strain's corresponding resistant spectrum have certain relevance and the same aggregation relationship.%目的：分析2011—2013年河南省肠炎沙门菌临床分离株的耐药状况与分子分型。方法收集2011年3月至2013年12月分离于河南省7个腹泻病多病原监测哨点医院的120株肠炎沙门菌，根据国际细菌性传染病分子分型监测网络公布的非伤寒沙门菌PFGE分型方法与美国临床与实验室标准协会沙门菌纸片法（Kirby-Bauer, K-B）药敏测试方案，对120株肠炎沙门菌进行8类抗生素的药敏测试与PFGE分子分型分析。结果120株肠炎沙门菌中，77株分离自男性，43株分离自女性；从0～5岁低龄儿童中共分离78株（65.0%），其中6月龄至2岁婴幼儿57株（47.5%）；分离时间集中于每年的5—10月份，其中3—4月份11株（9.2%），5—7月份48株（40.0%），8—10月份54株（45.0%），其余月份7株（5.8%），具有较典型的夏秋季节特征。120株肠炎沙门菌对氨苄西林的耐药率为50.0%（60株）；对头孢他啶的耐药率为14.2%（17株），对头孢噻肟的耐药率为18.3%（22株）；对头孢吡肟的耐药率为5.8%（7株）；对萘啶酸的耐药率为61.7%（74株）；对环丙沙星的耐药率为8.3%（10株），对诺氟沙星的耐药率为5.8%（7株）；对庆大霉素的耐药率为42.5%（51株），对链霉素的耐药率为21.7%（26株）；对氯霉素类抗生素的耐药率为30.0%（36株）；对甲氧苄氨嘧啶的耐药率为11.7%（14株），对复方磺胺甲唑的耐药率为71.7%（86株）；对四环素的耐药率为47.5%（57株）。120株肠炎沙门菌均对2种以上抗生素耐药，其中耐3～4种的为28株（23.3%），耐5～6种的为38株（31.7%），耐7～8种的为39株（32.5%）。经XbaⅠ酶切和PFGE电泳后，120株肠炎沙门菌共分为44种带型（EN1～EN44），各带型包含菌株数为1～35株不等，带型相似度在54.3%～100.0%之间，EN14、EN19分别包含35和29株菌，为该血清型主要优势带型。结论河南省临床分离的肠炎沙门菌耐药状况比较严重，PFGE带型具有优势带型特点的同时又与其对应的耐药谱具有一定的关联性与聚集性。

摘要： 目的：了解鲍氏志贺菌临床分离株毒力基因分布、耐药性、分子分型及菌株间流行病学相关性。方法从我院2015年6月—10月间门诊就诊腹泻病患者的粪便中分离收集9株鲍氏志贺菌。采用K-B纸片扩散法行抗生素药敏试验，聚合酶链反应技术检测毒力基因，脉冲场凝胶电泳技术及多位点序列分型确定分子分型，分析菌株间流行病学相关性。结果9株鲍氏志贺菌共分为3个血清亚型：Ⅰ型1株，Ⅱ型3株，Ⅳ型5株。9株菌均为多重耐药菌：对氨苄西林耐药率为9/9，对头孢他啶、链霉素、庆大霉素、复方新诺明、头孢噻肟、头孢曲松、四环素、诺氟沙星和左氧氟沙星的耐药率分别为1/9、4/9、4/9、4/9、5/9、5/9、6/9、6/9、6/9，未发现阿米卡星、头孢哌酮-舒巴坦和亚胺培南耐药株；ipaH的检出率为100%，sen、set1A、set1B、ial、virA、icsA、sigA的检出率均为0，pic、sepA、sat的检出率分别为4/9、5/9、7/9；9株鲍氏志贺菌通过脉冲场凝胶电泳共分为8种谱型，相似性在63.21%~100%。多位点序列分析结果显示6株为ST648，2株为ST131、1株为ST10。结论本院分离的9株鲍氏志贺菌共分3个血清亚型，多重耐药情况严重，菌株间亲缘关系相对较远，属非暴发病例。%Objective To understand genetic distribution, drug resistance, molecular typing and the epidemiological relativeness between strains of the Shigella boydii virulence. Methods Nine Shigella boydii strains were isolated form stool samples of patients with diarrhea from the Enteric Disease Clinic of the Second Hospital of Tianjin Medical University in June-October 2015. The strains were identified by biochemical test and serum agglutination test. Antibiotics susceptibility test was carried out using the Kirby-Bauer method. Polymerase chain reaction was used for detecting virulence genes. Pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST) technique were used to determine the epidemiological relationship between nine Shigella boydii strains. Results There were three subtypes in nine isolated Shigella boydii samples, including one, three and five isolates inⅠ,Ⅱ,Ⅳsubtypes respectively. All of the 9 isolates were multi-drug resistant. The resistant rate of these strains for ampicillin was 100%(9/9), and then the resistant rates of these strains for ceftazidime, streptomycin, gentamicin, trimethoprim/sulfamethoxazole, cefotaxime, ceftriaxone, norfloxacin and levofloxacin were 1/9, 4/9, 4/9, 4/9, 5/9, 5/9, 6/9, 6/9 and 6/9, respectively. All of these strains were sensitive to amikacin, cefperazone-sulbactam and imipenem. The ipaH was carried by all the testing strains, and none of the strains carried the sen, set1A, set1B, ial, virA, icsA and SigA. The detective rates of pic, sepA and sat were 4/9, 5/9 and 7/9 strains, respectively. Nine shigella boydii strains were divided into 8 PFGE types. The similarity between the spectrums of PFGE was 63.21%-100%. Multilocus sequence typing showed that six isolates were belonged to ST648, two isolates were ST131 and one isolate was ST10. Conclusion Nine isolates of Shigella boydii (divided into three subtyping) isolated from our hospital are multi-drug resistant and they have distant relationships, belonging to the dissemination of case.

摘要： 本发明涉及生物技术领域，特别涉及一种乳源革兰氏阳性菌的脉冲场凝胶电泳分型方法。该方法包括：将乳源革兰氏阳性菌裂解、DNA酶切后，采用脉冲场凝胶电泳技术对DNA片段进行分离，所述脉冲场凝胶电泳的初始转换时间为1.8～2.5s，终末转换时间为8～10s。本发明脉冲场凝胶电泳分型方法可对乳源革兰氏阳性菌进行准确分型。

摘要： 本发明公开了一种脉冲场凝胶电泳包被病原体胶块的切胶装置，该装置包括切胶平台、刀头和载胶件，所述刀头具有间距为2mm的刀片组件，无需人为对准，保证了切胶的准确性；切胶平台具有滤水结构，方便待切胶块和TE缓冲液的分离以及回收，载胶件与切胶平台配合使用，方便取出切好的小胶块，而且载胶件与切胶平台进一步引入分区的凸台后，构成多个独立的切胶区域，可以同时切割多个胶块，高效、快捷。本发明彻底改变了当前人工切割胶块误差大、速度慢的现状。

摘要： 本实用新型公开了一种脉冲场凝胶电泳后续实验用凝胶分切工具，包括定位框，所述定位框的下表面对称固定有连接板，且两个所述连接板的下端固定安装有防护壳，所述防护壳的下端安装有支腿，且支腿在防护壳的下表面均匀分布有四组，并且两个所述连接板之间卡合设置有导向面板，在原有脉冲场凝胶电泳后续实验用凝胶分切工具基础上，对现有的小刀切割方式进行了优化调整，采用了两组刀片配合的分切方式，将刀片对应套在两组微型转盘上，并通过镊子等工具依次旋转两组微型转盘，使得对应的卡块克服压紧条的摩擦阻力进行旋转，并且在压紧条逐渐增大的阻力作用下实现对卡块的定位，从而方便刀片的快速拆装。

摘要： 本实用新型涉及一种脉冲场凝胶电泳中凝胶的转移装置，它包括镂空平板(1)，所述镂空平板(1)由左右对称布置两块小平板(11)相互拼接而成，所述镂空平板(1)左右两侧设置有手柄(2)。本实用新型一种脉冲场凝胶电泳中凝胶的转移装置，能够实现琼脂糖凝胶的无破损转移，其实用性强，操作简单，可以减少胶块转移过程中对胶块的损伤，降低转移胶块的难度，从而提高操作效率，加快实验的速度。

摘要： 本实用新型公开了一种脉冲场凝胶电泳凝胶染色装置，包括盒体和防滑条，所述盒体的表面与防滑条内壁的一侧固定连接，所述盒体的底部固定连接有调节筒，所述调节筒内壁的两侧均固定连接有三角块，所述三角块的表面开设有卡槽，所述三角块的表面活动连接有支撑杆，本实用新型涉及凝胶染色技术领域。该脉冲场凝胶电泳凝胶染色装置，使得装置可以方便的调节左右两侧的高度，保证了装置在不水平的摇床表面可以通过稳定快速调节达到整体水平的效果，并且通过连接杆、辅助盒和弹性片的联合设置，使得通过支撑杆转动调节装置高度后，可以通过弹性片的弹性保证装置中的支撑杆始终与三角块紧密卡合，从而装置保持稳定。

摘要： 一种脉冲场凝胶电泳凝胶染色装置，其特征在于，它包括盒体、盒盖和密封塞；所述的盒体的侧壁外侧有一对把手，盒体的底部下面均匀布置若干个吸盘，盒体的一个侧壁下方设有连通盒体内的凸出的管口，密封塞塞紧并密封管口的通透孔；盒盖通过密封圈和卡扣盖紧密封盒体。本实用新型提供的染色盒的盒体与盒盖密封紧密，减少了染液的漏出；侧壁下方的出水孔方便染液倾倒；底部的吸盘旋钮保证盒子水平；盒体两侧的把手方便实验操作。这些措施可以减少实验误差，提高不同实验菌株脱氧核糖核酸脉冲场凝胶电泳分离后的成像效果，使菌株之间分子指纹图谱分析的平行性和可比性都显著提高。

摘要： 本实用新型公开脉冲场凝胶电泳包埋胶块的切胶器，包括切胶板，所述切胶板内部设有数个矩形凹槽，所述固定支架表面设有刻度线，固定支架的左右两侧分别设有第一切胶平台和第二切胶平台，所述第一切胶平台固定设置在固定支架上，其第二切胶平台活动连接在固定支架上，所述第一切胶平台和第二切胶平台均包括切割刀片和手持端部。有益效果在于，矩形凹槽的设置可使得对胶块进行固定，便于其切割，并通过切胶平台的设置，可进行长度调整，矩形凹槽上设有编号，与电泳时候的胶块位置一致，保证上样的准确性；结构简单，可提高切割效率，节省时间；切胶平台的整体性的结构设置，使其切出来的胶条两边光滑，利于电泳成图效果，便于后续分析。

摘要： 本实用新型公开了一种脉冲场凝胶电泳包被病原体胶块的切胶装置，该装置包括切胶平台、刀头和载胶件，所述刀头具有间距为2mm的刀片组件，无需人为对准，保证了切胶的准确性；切胶平台具有滤水结构，方便待切胶块和TE缓冲液的分离以及回收，载胶件与切胶平台配合使用，方便取出切好的小胶块，而且载胶件与切胶平台进一步引入分区的凸台后，构成多个独立的切胶区域，可以同时切割多个胶块，高效、快捷。本实用新型彻底改变了当前人工切割胶块误差大、速度慢的现状。

摘要： 本实用新型公开了一种脉冲场凝胶电泳清洗装置，解决了现有技术中plug的清洗存在耗时长、操作繁琐的问题。本实用新型包括清洁管体，所述清洁管体顶端开口，底端设置有进口管和出口管，所述清洁管体上还设置有通过开口位置处伸入清洁管体内的滤水勺；所述清洁管体的数量设置为多个，依次连接为一体结构；相邻两个清洁管体通过进口管和出口管相互连通。本实用新型通过合理的结构优化，极大地节省了操作时间，防止plug转移的损坏，并且有效避免编号错误，提高实验效率。

摘要： 本实用新型公开了一种脉冲场凝胶电泳细菌DNA包埋胶块的切割装置，包括平台、方形板、支架和切胶刀，方形板设置在平台上，并可沿平台水平直线滑动，平台上沿方形板的滑动方向设置有刻度线，方形板两端的上端横向竖直且相对设置有前挡料板和后挡料板，前挡料板和后挡料板之间放置待切割的胶块，后挡料板可沿方形板朝向前挡料板滑动靠近或远离，切胶刀通过支架竖直固定在平台上，并位于方形板的滑动轨迹的上方，切胶刀可相对于支架上下移动，以将方形板上的待切割的胶块切割成所需长度。本实用新型提供一种可快速且准确对多条胶块进行切割的脉冲场凝胶电泳细菌DNA包埋胶块的切割装置。