多位点酶电泳法在菌痢暴发中的应用

从广州、江门两地分离到７９株Ｆ２ａ，采用多位点酶电泳（ＭＥＥ）技术对该菌的６种代谢酶进行了检测。结果表明，７９株Ｆ２ａ共分为１０个电泳型（ＥＴ），存在一个优势型（ＥＴ１）和一个次优势型（ＥＴ２），以及８个非优势型。该现象说明，ＥＴ１型是引起菌痢暴发和流行的优势克隆。用ＭＥＥ方法检测表型一致的Ｆ２ａ，可发现二者之间具有差异。     

福氏2a志贺菌肠毒素ShET1/ShET2基因分型在菌痢暴发研究中的应用

目的 对福氏2a志贺菌肠毒素ShET1(Shigella enterotoxinl)和肠毒素ShET2(Shigella enterotoxin2)进行基因分型，提高菌痢暴发流行时同源克隆的鉴定分析水平。方法 对在菌痢暴发期间分离的8株福氏2a志贺菌用PCR法检测志贺菌肠毒素ShET1/ShET2基因，进行基因分型和同源克隆鉴定。结果 8株福氏2a志贺菌可分为3种基因型，即2株ShET1(-)/ShET2( ),1株ShET1( )/ShET2(-)，5株ShET1( )/ShET2( )。结论 本次菌痢暴发部分患可能并非真正的暴发病例，之所以被归入暴发事件之中，可能是一种与暴发事件在空间、时间上偶合的散发病例。     

菌痢暴发的分子流行病学分析

分析１９９２、１９９４年广州、江门两地菌痢暴发期间分离的８４株福氏２ａ志贺氏菌（广州暴发株３０株、散发株２８株、江门暴发株１２株、病例接触者分离株１４株）的质粒图谱及质粒ＤＮＡ限制性酶切图谱特征。结果表明：发生于不同地点、不同时间且传染来源不相同的两起菌痢暴发，由福氏２ａ志贺氏菌的同一克隆引起，该克隆可能是广东地区福氏２ａ志贺氏菌的优势克隆。江门菌痢暴发期间，自密切接触者分离的同型菌株分属３个不同的克隆，提示部分病例接触者感染的福氏２ａ志贺氏菌并非来自病例。比较表型分析（生化反应、血清学分型）与质粒分析，证实表型分析难以鉴别来源不同的同型克隆，而质粒分析提供了简便、敏感、特异鉴定不同来源同型克隆的有效手段。     

PCR检测技术在志贺菌引起的腹泻疫情中的应用

目的：通过采用PCR方法对某次疫情中分离的福氏1 a志贺菌进行保守基因检测及毒力基因分析研究,探讨在感染性腹泻疫情中,PCR技术快速检测志贺菌的应用。方法：对标本中分离的7株福氏1 a志贺菌,用PCR方法检测侵袭性质粒抗原基因（ipaH）、志贺肠毒素1（ShET-1）和志贺肠毒素2（ShET-2）的基因检测,并进行基因分型。结果：7株F1 a菌均检测出ipaH基因,ShET-1和ShET-2,基因型为ShET1（＋）/ShET2（＋）,整个实验可在3 h内完成。结论：该所建立的PCR检测方法准确、快速、简便、特异性强,为其应用于环境、食品及临床腹泻样本中志贺菌的检测打下良好基础。ShET-1/ShET-2基因分型对福氏志贺菌暴发流行中的同源性分析有重要意义。     

1998年沈阳地区从业人员痢疾杆菌的带菌率及菌型分布调查

志贺氏菌属（Ｓｈｉｇｅｌｌａ）感染在我国仍居腹泻病因的首位,在人群中传染源数量甚多,而且轻型症状称无症状带菌占比例较高。但目前关于沈阳市健康人群体检带菌情况及菌型分布尚没有报道。现就１９９８年沈阳地区从业人员健康体检结果报告如下。材料与方法　（１）样品来源：沈阳地区从业人员健康体检１６４６３０份。（２）方法：肛拭（采便棒）３～４ｃｍ深度旋转半圈,直接分离于ＳＳ和ＥＭＢ琼脂平板各一块,３７℃,１８～２４ｈ培养。挑取可疑菌落穿三糖铁斜面,３７℃１８～２４ｈ培养,符合志贺氏菌属特征者接种生化培养基进…     

济源市福氏志贺菌毒力基因分型与分析

目的：对河南省济源市2002—2005年分离之福氏志贺菌株进行基因分型和分析。方法：应用多重PCR方法检测志贺菌肠毒素1基因（ShET-1B）、志贺菌肠毒素2基因（ShET-2）、侵袭性质粒抗原H基因（ipaH）和侵袭相关基因（ial），根据产物分子量定性判断结果。结果：90株福氏志贺菌具有2种毒力基因型，ShET-1阳性率达到98．9％。结论：济源市福氏志贺菌毒力基因型具有高度的一致性。     

志贺菌毒力基因的多重PCR鉴定

目的：建立志贺菌肠毒素1基因（ShET-1B）、志贺菌肠毒素2基因（ShET-2）、侵袭性质粒抗原H基因（ipaH）和侵袭相关基因（捌）的多重PCR鉴定方法，实现志贺菌多基因同时检测。方法：选择志贺菌的4种毒力基因作为靶基因，应用Touchdown PCR方法，在一个反应管中同时进行扩增，根据产物分子量定性判断结果。结果：多重PCR同时检测与单基因PCR分别检测4种毒力基因两种方法具有相同的灵敏度与特异性。624株志贺菌具有6种毒力基因型，571株福氏志贺菌（包括11个血清型）中556株ShET-1B阳性。结论：多重PCR鉴定志贺菌毒力基因具有简便、快速的特点，适用于毒力基因鉴定和流行病学调查研究。ShET-1B基因可能广泛存在于福氏志贺菌之中。     

三基因PCR检测产毒性大肠杆菌和志贺菌的研究

三基因ＰＣＲ检测产毒性大肠杆菌和志贺菌的研究赵敏俞守义王红王雅贤吴敏在发展中国家，每年大约有五百万左右的儿童和婴儿死于急性腹泻［１］。这种疾病的致病因素主要是产毒性大肠杆菌（ＥＴＥＣ）和志贺菌。ＥＴＥＣ在生长繁殖过程中释放出耐热肠毒素（ＳＴ）和不耐热...     

２００４－２０１４年宁波市细菌性痢疾流行特征及 病原学分析

摘要：目的　了解宁波市细菌性痢疾流行特征, 为防控工作提供依据。方法　采用ＳＰＳＳ１７．０ 和Ｅｘｃｅｌ ２００３软件,对宁波市２００４－２０１４年细菌性痢疾疫情资料进行数据统计处理,采用Ｐｅａｒｓｏｎχ ２ 检验,犘＜ ０．０５为差异有统计学意义,分离和收集志贺菌进行鉴定分型、耐药性和致病力检测。结果　宁波市细菌性 痢疾年发病率在１．７８／１０万～４２．５９／１０万,平均为１８．６９／１０万。全年均可发病,发病高峰为８～９月。志 贺菌流行株以Ｂ群志贺菌为主,Ｄ 群志贺菌为次,呈福氏２ａ血清型、福氏ｃ亚型和宋内氏志贺菌格局。菌 株耐药严重,多为多重耐药株;携带毒力基因种类多、毒性强。结论　宁波市细菌性痢疾发病率逐年下降, 已维持在较低水平,但菌株的血清型在转换,耐药性和毒力显示其有较强的致病性,引起暴发与流行的因 素仍未改变,提示应加强细菌性痢疾的监测与防控。 关键词：细菌性痢疾;流行病学;病原学;耐药性;毒力 中图分类号：Ｒ５１６．４　　文献标识码：Ａ　　文章编号：１００９ ６６３９ （２０１７）０５ ０３５５ ０６     

1985—1992年广西志贺氏菌菌型分布及耐药性的实验研究

本文分析了广西１４个地、市、县的１２８９株志贺氏菌的群、型，Ｂ群志贺氏菌是我区的主要流行株，占６５．６３％，分布于１４个地、市、县，其中Ｆ２ａ居首，占２４．９０％。对７８６株志贺氏菌作了１６种抗生素的耐药性监测，本区的痢疾菌对ＴＣ、ＳＭ、ＣＯＳ、ＳＤ的耐药率分别高达８７．７９％、６６．６７％、６６．１６％、和８２．１９％。     

Molecular epidemiology of Shigella infection in Central Australia

Shigellosis is endemic in Central Australia and the infections are predominantly due to Shigella flexneri 6, Shigella flexneri 2a and Shigella sonnei. Plasmid profiles of isolates collected from 1985-9, suggested that infections caused by Shigella flexneri 6 were predominantly due to a single clone, whereas those caused by Shigella flexneri 2a and Shigella sonnei were due to several genetically diverse strains, although strains with identical plasmid profiles were found in widely separated geographical areas and in different years.     

福氏志贺菌4c型暴发和散发菌株的脉冲场凝胶电泳分型

目的了解杭州地区福氏志贺菌4c型暴发菌株和散发菌株的分子分型特征。方法对2003和2005年杭州地区的2次食物中毒暴发分离的福氏志贺菌菌株（暴发1和暴发2,菌株数分别为13株和12株）和2005年1例散发腹泻患者分离的福氏志贺菌4c型离株（1株）进行生化鉴定和血清分型,PCR检测侵袭性抗原基因ipaH,改良Kirby-Bauer纸片法检测菌株的耐药谱及脉冲场凝胶电泳（pulse field gel electrophoresis,PFGE）进行分子分型。结果暴发1中分离的13株菌株均为福氏4c型,但检测的6株菌株间的PFGE图谱存在着相当的差异,Dice系数为0．78至0．92。暴发2中分离的菌株有10株福氏4c型和2株福氏x型,所有菌株的PFGE图谱几乎完全一致。暴发2分离株和暴发1的部分分离株可能存在着一定相关性（Dice系数大于0．8）,而散发菌株与2起暴发的绝大部分菌株间的距离较远（Dice系数小于0．8）。暴发1、暴发2的分离株和1株散发菌株对14种抗生素的耐药谱略呈差异,暴发2中分离的福氏4c型和x型菌株的耐药谱一致。结论PFGE能够很好地辨别福氏志贺菌4c型的两起暴发菌株和1株散发菌株。福氏志贺菌4c型菌株具有易变性,可在暴发流行过程中产生PFGE图谱的差异、血清亚型的转换、耐药谱的变化等。     

Prevalence of the genes for shigella enterotoxins 1 and 2 among clinical isolates of shigella in Israel

Two enterotoxins, shigella enterotoxin 1 (SHET1) and shigella enterotoxin 2 (SHET2) have been recently characterized and are believed to play a role in the clinical manifestation of shigellosis. One hundred and twenty-one isolates of Shigella spp. of 13 different serotypes and variants and 10 isolates of enteroinvasive Escherichia coli (EIEC) isolated in Israel, were examined by polymerase chain reaction for the presence of SHET1 and SHET2 genes. SHET1 was only prevalent among isolates of S. flexneri 2a while SHET2 was found in all the serotypes that were tested except for several isolates of S. flexneri 1b that lost their virulence plasmid during storage. In addition, we found that the S. flexneri 2a vaccine strain T-32 Istrati contains the gene encoding for SHET1 but not that encoding for SHET2, suggesting that the latter is located within a large deletion occurring in the 140 Mda plasmid of this S. flexneri 2a non-invasive vaccine strain.     

福氏志贺菌4c亚型的分子生物学特征分析

目的 了解杭州地区2003-2007年从爆发和散发细菌性痢疾患者中分离之福氏志贺菌4c亚型(F4c)菌株的分子生物学特征.方法 对24株F4c菌株以生化反应和血清分型进行鉴定,利用PCR法检测侵袭性质粒抗原H基因(ipaH)及脉冲场凝胶电泳(PFGE)进行分型分析.结果 24株菌株全部为F4c,均具有ipaH毒力基因.用PFGE分型方法将24株菌株分成17种不同的带型,Dice系数为0.71～1.00.2005年、2006年、2003年三次爆发(分别简称为爆发1、2、3)中各自分离到的菌株基因型紧密相关.三次爆发中的部分菌株可能存在一定的相关性(Dice系数>0.8).散发菌株和爆发1、3菌株间的距离较远(Dice系数<0.8),和爆发2菌株间可能存在一定的相关性(Dice系数>0.8).结论 F4c菌株具有ipaH毒力基因,PFGE提示杭州地区近年F4c三起爆发菌株可能属共同的克隆群.     

Immunogenicity and characterization of WRSF2G11: a second generation live attenuated Shigella flexneri 2a vaccine strain

Recent clinical trials involving live attenuated Shigella vaccine strains SC602 and WRSS1 have revealed that deletion of the virG(icsA) gene dramatically reduces virulence in human volunteers. These strains can be given at low oral doses and induce a strong, and in some cases, protective immune responses. However, residual vaccine associated reactogenicity suggests that further attenuation is required. A recent clinical trial indicated that the set and sen enterotoxin genes contribute to the symptoms of fever and diarrhea observed with live Shigella vaccine strains. Based on these findings, a Shigella flexneri 2a vaccine candidate, WRSf2G11, with deletions in the virG(icsA), set and sen genes has been constructed using the lambda red recombinase system. The immunogenicity and protective efficacy of WRSf2G11 compares favorably with SC602 following either intranasal (IN) or ocular (OC) immunization of guinea pigs. Taken together, these data indicate that second generation virG-based Shigella vaccine strains which lack enterotoxin genes, such as WRSf2G11, will likely show lower levels of reactogenicity without hampering the robust immune responses achieved with previous live vaccines.     

Electropherotyping of plasmid DNA of different serotypes of Shigella flexneri isolated in Bangladesh

One hundred and twenty-five Shigella flexneri strains, isolated during January-December 1984, at the Dhaka treatment centre of the International Centre for Diarrhoeal Disease Research, Bangladesh, were serotyped using absorbed rabbit antisera specific for all type- and group-factor antigens, as well as a group of ten mouse and rat monoclonal antibodies. Electropherotypes of the plasmid deoxyribonucleic acid (DNA) were also determined. S. flexneri 2a was the predominant serotype followed by 3b, 1a, and 2b. The recently described E1037 antigen was also found in three strains of S. flexneri serotype 6. Electropherotyping of the plasmid DNA showed that three plasmids of approximately 140, 2.7, and 2 megadalton (MDa) were present, respectively, in 97, 97 and 94% of the 125 strains. Additional plasmids of various other sizes were also present in different serotypes except in serotype 2a. The additional plasmids again appeared to be specific for that particular serotype. For example, all 12 strains of S. flexneri 2b harboured an additional plasmid of approximately 1 MDa. Thus, electropherotyping of plasmid DNA of different serotypes of S. flexneri might be useful to differentiate S. flexneri from other species of Shigella and in identifying different serotypes of S. flexneri. Therefore, the common plasmids, plus the additional plasmids, could be used to identify epidemic, as well as sporadic, subclones of S. flexneri strains.     

郑州地区农村83株痢疾杆菌菌型分布和耐药性调查

对1981年在郑州郊区检出的83株痢疾杆菌作了菌型分布与耐药性调查，志贺氏Ⅰ型66株，占79.5%；福氏志贺氏菌14株，占16.9%，宋内氏志贺氏菌2株，占2.4%；鲍氏志贺氏菌1株，占1.2%。认为志贺氏I型为当地优势菌种，与当地近年来菌群菌型分布明显不同。其中67株痢疾菌作了药敏感试验，对庆大霉素、卡那霉素、新霉素、呋喃妥因和痢特灵较敏感。但对常用抗菌药物金霉素、土霉素、磺胺嘧啶耐药高达92.5~100%。由于对常用抗菌药物耐药性逐渐增强多种耐药菌株广泛存在，给菌痢防治带来一定困难，因此对耐药性的动态变化应予充分重视。