食品中沙门菌整合子与耐药性关系研究

目的：研究食品中沙门菌的整合子携带情况及整合子与耐药性之间的关系。方法：PCR检测细菌总DNA中1、2、3类整合酶基因，确定细菌携带整合子的情况。整合子阳性菌档进一步检测整合子的可变区，分析插入基因盒的序列。结果：69株大肠菌中25株携带1类整合子，集中分布在对4种以上抗生素耐药的菌株中。插入基因盒主要是dfr(甲氧苄氨嘧啶)和aadA(氨基糖甙类)基因盒，各种基因盒组合中最常见的是difrl—aadA1。结论：细菌的多重耐药性与整合子携带高度一致，但是单个菌株的耐药谱与整合子的耐药基因盒缺乏对应关系。     

食品中大肠埃希菌整合子与耐药性的关系研究

目的：研究食品中大肠埃希菌的整合子携带情况及其与耐药性之间的关系。方法：PCR检测细菌总DNA中1、2、3类整合酶基因，确定细菌携带整合子的情况。阳性菌株进一步检测整合子的可变区，分析插入基因盒的序列。结果：50株大肠杆菌中19株携带1类整合子，2株携带2类整合子，集中分布在对4种以上抗生素耐药的菌株中。插入基因盒主要是dfr和aadA类基因盒，各种基因盒组合中最常见的是dfr17-aadA5。结论：细菌的多重耐药性与整合子携带高度一致，但是单个菌株的耐药谱与整合子的耐药基因盒缺乏对应关系。     

健康人肠道大肠埃希菌整合子和耐药基因盒的研究

目的：研究健康人肠道大肠埃希菌的整合子携带情况与整合子携带的基因盒种类及其与耐药性之间关系.方法：PCR检测Ⅰ类整合子,确定细菌携带整合子的情况.整合子阳性菌株进一步检测整合子的可变区,分析插入基因盒的序列.结果：150株大肠埃希菌中有38株携带Ⅰ类整合子,集中分布在对3种以上抗生素耐药的菌株中.插入基因盒集中在甲氧苄氨嘧啶耐药基因（dfr）和氨基糖甙类抗生素耐药基因（aadA）.结论：细菌的多重耐药性与整合子携带高度一致,但是细菌耐药谱和其整合子无对应关系.     

南通地区食源性沙门菌多重耐药性与整合子的相关性研究

目的通过对南通地区食源性沙门菌整合子的检测及耐药性分析,初步探讨整合子、基因盒与耐药性的相关性。方法采用纸片扩散法(kirby-bauer)对67株食源性沙门菌进行耐药性分析,从中选取多重耐药性菌株,提取纯化不同菌株共有的质粒,应用PCR法扩增整合子并进行分类;随机选取1例PCR产物进行DNA测序。结果 67株南通地区食源性沙门菌中31.3%具有多重耐药性;Ⅰ类整合子阳性14株,未发现Ⅱ类、Ⅲ类整合子;10株沙门菌携带质粒,质粒数目为1个~3个,大小为0.5 kb~4 kb;Gen Bank数据库BLAST软件分析1例PCR产物(1.5 kb片段)含有3个开放阅读框,分别与dfr A1、sat2和aad A1耐药基因盒对应。结论南通地区食源性沙门菌对氨苄西林、环丙沙星和哌拉西林等药物出现较高的耐药性;耐药基因可由Ⅰ类整合子携带,并可通过接合试验发生转移,这对细菌多重耐药性的产生和传播转移起到非常重要的作用。     

整合子及其相关基因盒在临床产ESBLs肺炎克雷伯菌的分布

目的检测Ⅰ~Ⅲ类整合子及Ⅰ类整合子相关基因盒在产ESBLs肺炎克雷伯菌临床分离株中的分布,分析整合子对细菌耐药性的影响。方法用K-B纸片扩散法对108株产ESBLs肺炎克雷伯菌临床分离株进行药敏试验;采用聚合酶链反应(PCR)分析108株产ESBLs肺炎克雷伯菌株的Ⅰ~Ⅲ类整合子;对Ⅰ类整合子阳性菌进行整合子相关基因盒检测。结果108株菌中有58株(53.70%)含有Ⅰ类整合子,未检测到Ⅱ类整合子及Ⅲ类整合子阳性菌;在Ⅰ类整合子阳性菌中,有52株携带Ⅰ类整合子相关基因盒,占89.66%;分离自同一科室的部分菌株携带大小相同的基因盒;Ⅰ类整合子阳性菌株的耐药率高于整合子阴性的菌株。结论Ⅰ类整合子及整合子相关基因盒在产ESBLs肺炎克雷伯菌临床分离株中分布广泛,整合子在细菌耐药中发挥作用。     

尿液分离大肠埃希菌Ⅰ类整合子分子特征及其与耐药性的关系

目的 了解尿液分离大肠埃希菌中Ⅰ类整合子及相关基因盒的分布,分析整合子与细菌耐药性的关系.方法 选取108株临床非重复分离自尿液标本中的大肠埃希菌,经全自动细菌分析系统鉴定并检测其对临床常用抗菌药物的耐药性,聚合酶链反应(PCR)检测Ⅰ类整合酶基因,对Ⅰ类整合酶阳性菌株用PCR扩增并测序分析可变区基因盒种类及可变区启动子类型.结果 108株临床非重复分离自尿液大肠埃希菌中有60株占56.0％检测到Ⅰ类整合酶基因;共检测出5种不同长度的可变区片段:8株750 bp片段不含基因盒,2株1500 bp片段为aadA2,34株2200 bp片段为dfrA 17-aadA5,4株2500 bp片段为dfrA 12-orfF-aa dA2,2株2800 bp片段为aacA4-cmlA1,其中1株大肠埃希菌同时扩增出2200、2500 bp的可变区片段,有11株Ⅰ类整合酶基因阳性大肠埃希菌未扩增出可变区;可变区启动子大多为相对较弱的启动子PcH1、PcW;Ⅰ类整合子阳性菌株对多种抗菌药物的耐药率高于Ⅰ类整合子阴性菌株.结论 Ⅰ类整合子及相关基因盒在尿液分离大肠埃希菌中分布广泛,与细菌耐药性关系密切,所携带的整合子具有较强的捕获外源性耐药基因盒的能力.     

健康人群大肠埃希菌Ⅰ类整合子与耐药性关系

目的 研究健康人群大肠埃希菌中Ⅰ类整合子携带情况及分子特征,分析其与细菌耐药性关系.方法 采用Vitek32细菌分析仪和纸片扩散法,检测150株大肠埃希菌抗生素敏感性,PCR检测总DNA中Ⅰ类整合子,扩增产物进行限制性核酸内切酶分析,测序确定携带基因盒种类,脉冲场凝胶电泳对整合子阳性菌株分子分型.结果 150株大肠埃希菌中,83株菌耐受>3种抗生素,耐多药率55.3%;超广谱β-内酰胺酶(ESBLs)阳性5株,阳性率3.3%;携带Ⅰ类整合子42株,扩增子150～3 000 bp,,有8种带型;耐药基因盒主要是dfr和aadA类,最常见基因盒排列dfr17～aadA5;整合子分布在不同基因型菌株中.结论 健康人携带的大肠埃希菌中广泛存在Ⅰ类整合子,与细菌耐多药性密切相关;分子分型结果提示整合子能在细菌种内水平传播,整合子是介导细菌耐药性的重要分子机制.     

致泌尿系统感染中大肠埃希菌Ⅰ类整合子的研究

目的：分析泌尿系统感染中大肠埃希菌Ⅰ类整合子的流行情况和分子特征。方法：应用全自动细菌分析仪Microgcan WalkAway40和纸片扩散法，对40株大肠埃希菌进行抗生索敏感性测定，PCR扩增细菌总DNA上的Ⅰ类整合子。对扩增产物进行测序。并分析其中的基因盒。利用脉冲场电泳（PFGE）对菌株进行分子分型。结果：22株细菌含有Ⅰ类整合子，整合子大小为1000、1200、1700和3000bp，22株细菌各含1-2个整合子。1000bp整合子含基因盒aadA1。1200bp整合子含基因盒dfr1—OFRx，1700bp整合予含基因盒dfr17-aadA5，3000bp整合子两端分别含基因盒aadB、cmlA1，40株细菌被分为不同的基因型，在不同基因型菌株中发现了相同的整合子。结论：整合子在泌尿系统感染大肠埃希中广泛存在，整合子是介导细菌耐药性的重要分子机制，基因分型结果提示整合子在细菌种内水平传播。     

佛山市沙门菌多重耐药及第Ⅰ类整合子携带情况分析

目的 了解佛山市沙门菌的多重耐药以及第I类整合子的携带情况,为研究沙门菌耐药机制提供参考。方法 收集2015—2017年佛山市食源性疾病监测中分离的390株沙门菌作为本研究的样本菌株,按照Kauffman-White的分类标准进行沙门菌血清分型,采用半定量药敏检测法测定沙门菌对14种抗生素的最小抑菌浓度（ MIC）,采用聚合酶链反应（PCR） 检测第I类整合子相关的整合酶intI基因。结果 390株沙门菌共分为9个血清群、43个血清型,其中以B群的鼠伤寒沙门菌变种（I4,5,12：i：-）检出数最多（136株,占34.87%）。药敏试验结果显示,耐药率最高的是氨苄西林,耐药率为69.23%,其次为四环素,耐药率为67.95%。390株沙门菌中共检出329株耐药菌株,耐药率为84.36%;其中多重耐药菌株252株,多重耐药率为64.62%（252/390）。37.95% （148/390） 的沙门菌检测出第Ⅰ类整合子,54.37%（137/252）多重耐药菌株检出第Ⅰ类整合子。结论 佛山市沙门菌血清型别多样,耐药率及整合子携带率均较高,提示应从基因水平对沙门菌耐药性及其传播机制进行监测 。     

Ⅰ类整合子与铜绿假单胞菌多重耐药相关性研究

目的了解Ⅰ类整合子在临床分离的铜绿假单胞菌多重耐药株及敏感株中分布的差异,初步阐明Ⅰ类整合子是否与铜绿假单胞菌多重耐药性形成相关。方法应用聚合酶链反应（PCR）法分别检测临床分离的67株多重耐药铜绿假单胞菌和62株敏感株中Ⅰ类整合子的阳性率,并进一步用PCR证实Ⅰ类整合子阳性菌中整合子相关基因盒的存在。结果铜绿假单胞菌多重耐药株中,32株（47.76%）为Ⅰ类整合子阳性;敏感株中仅1株（1.61%）为Ⅰ类整合子阳性,多重耐药株的Ⅰ类整合子分布显著高于敏感株（χ2=36.02,P〈0.05）。PCR进一步证实多重耐药铜绿假单胞菌携带Ⅰ类整合子基因盒,而敏感株携带空整合子。结论多重耐药铜绿假单胞菌具有较高的Ⅰ类整合子阳性率,Ⅰ类整合子可能与铜绿假单胞菌的多重耐药性相关。     

Antimicrobial resistance in isolates of Salmonella spp. from pigs and the characterization of an S. Infantis gene cassette

One hundred and seven Salmonella isolates of various serotypes were investigated for resistance to a panel of nine antimicrobial agents by standardized methods. Thirty four isolates were susceptible to the selected antimicrobial agents. Thirty-six (of 107) were resistant to three or more antimicrobial agents and defined as multidrug resistant (MDR). Salmonella Typhimurium was the most resistant serotype. All resistant isolates were examined for the presence of class 1 integrons. Thirty-two integron-associated gene cassettes (of varying sizes) were identified. A 1,000-bp amplicon similar to that flanking the distal region of the Salmonella Genomic Island (SGI)-1 in Salmonella Typhimurium was detected in a majority of the S. Typhimurium isolates in this study. In contrast, a 1,800-bp amplicon was identified in all Salmonella Infantis isolates. This amplicon was completely characterized in one isolate. The presence of class 1 integrons in Salmonella spp. in pigs may be important if these zoonotic pathogens were to enter the food chain.     

High Prevalence of Class 1 to 3 Integrons Among Multidrug-Resistant Diarrheagenic Escherichia coli in Southwest of Iran

Objectives

Horizontal transfer of integrons is one of the important factors that can contribute to the occurrence of multidrug-resistant (MDR) bacteria. This study aimed to determine the prevalence of integrons among MDR Escherichia coli strains isolated from stool specimens and investigate the associations between the existence of integrons and MDR properties in the southwest of Iran.

Methods

There were 164 E. coli strains isolated from January 2012 to June 2012. Fecal specimens identified as E. coli by the conventional methods. Subsequently the antibiotic resistance was assessed using Clinical and Laboratory Standard Institute criteria. The presence of class 1–3 integrons and embedded gene cassettes was verified using specific primers by multiplex polymerase chain reaction assay.

Results

Among a total of 164 studied samples, 69 (42.07%) isolates were multidrug resistant. Class 1 and class 2 integrons were present in 78.26% and 76.81% MDR isolates, respectively. For the first time in Iran, class 3 integron was observed in 26.09% MDR isolates. Significant correlations were identified between: class 1 integron and resistance to amikacin, gentamicin, chloramphenicol, ampicillin, tetracycline, nalidixic acid, and co-trimoxazole; class 2 integron and resistance to aminoglycosides, co-trimoxazole, cefalexin, ampicillin, and chloramphenicol; and class 3 integron and resistance to gentamicin, kanamycin, and streptomycin.

Conclusion

Our results indicate that integrons are common among MDR isolates and they can be used as a marker for the identification of MDR isolates. Therefore, due to the possibility of a widespread outbreak of MDR isolates, molecular surveillance and sequencing of the integrons in other parts of the country is recommended.     

Occurrence and distribution of various genetic structures of class 1 and class 2 integrons in Salmonella enterica isolates from foodborne disease patients in Korea for 16 years

We have investigated the distribution of integrons among 752 multidrug-resistant Salmonella isolates from human febrile and/or diarrheal patients during 1992-2007 and analyzed their genetic characteristics. Here, we report extensive integron analysis results within human isolates during the last 16 years. The gene or gene cassette(s) in the class 1 integrons found in the isolates were dfrA7, dfrA12-orfF-aadA2, aadA2, bla(PSE1), dfrA1-aadA1, dfrA17-aadA5, bla(OXA1)-aadA1, aadB-aadA1, aadA22, aadA1, and aac6'Ib-bla(OXA1)-aadA2. Class 2 integrons harboring dfrA1-sat2-aadA1 gene cassette were also found in four isolates. Twenty-nine isolates including one Salmonella Schleissheim isolate had two integrons harboring aadA2 and bla(PSE1) in their variable regions of 1.0 and 1.2?kb amplicons, respectively, which have been also found in Salmonella genomic island 1 (SGI1) of multidrug-resistant Salmonella Typhimurium DT104. The presence of SGI1 in Salmonella Schleissheim isolate was proved by SGI1-specific polymerase chain reaction. We first report a Salmonella Schleissheim having SGI1, Salmonella Typhimurium and Salmonella Heidelberg having the class 2 integron with dfrA1-sat2-aadA1 cassettes, Salmonella London with the aac6'Ib-bla(OXA1)-aadA2 gene cassette, Salmonella Chailey with the gene cassette of aadA22, and coexistence of two class 1 integrons carrying aadA22 and dfrA12-orfF-aadA2 in Salmonella Typhimurium.     

菌落杂交法鉴定沙门菌第一类整合子

目的:建立一种快速而经济的鉴定细菌中整合子的新方法,分析整合子介导的细菌耐药性。方法:以第一类整合酶基因intI1为探针,采用地高辛进行标记,菌落杂交的方法来鉴定第一类整合子阳性菌株。结果:发现在23株沙门菌中,四株第一类整合子阳性菌株,与采用第一类整合酶基因intI1 PCR扩增方法所得到的结果相一致。结论:菌落杂交法可以简单快速地鉴定细菌中整合子,这对研究整合子的特性及整合子在细菌耐药性中的介导作用具有重要意义。     

致血液系统感染大肠埃希菌中Ⅰ类整合子参与耐药的研究

目的：探讨致血液系统感染大肠埃希菌中Ⅰ类整合子介导耐药性的分子机制。方法：应用全自动细菌分析仪Microscan WalkAway40和纸片扩散法，对16株大肠埃希菌进行抗生素敏感性测定，PCR扩增细菌总DNA上的Ⅰ类整合子，对扩增产物测序并分析其中的基因盒。结果：8株细菌含有Ⅰ类整合子，整合子大小分别为600bp、1700bp和2500bp，600bp整合子含基因盒dfr2d，1700bp整合子含基因盒dfr17-aadm5。结论：整合子在介导细菌耐药性方面发挥着重要作用。     

Serotypes, antibiotic resistance, and class 1 integrons in Salmonella isolates from pediatric cases of enteritis in Tehran, Iran

The present study was conducted to investigate serotype distribution, antimicrobial resistance patterns, carriage of class 1 integron, and clonality of Salmonella strains isolated from patients aged 0-12 years in Tehran, Iran, during 2007-2008. A total of 139 Salmonella isolates were studied. Salmonella serotypes Enteritidis, Infantis, and Typhimurium included 84.9% of isolates, Enteritidis accounting for 41.7%. The most prevalent resistances were to doxycycline (64.7%), nalidixic acid (61.2%), tetracycline (51.8%), and streptomycin (42.8%). Fifty-three (38.1%) isolates contained class 1 integron. Eight different gene cassettes were identified, aadA1 being the most frequently encountered. Pulsed-field gel electrophoresis showed that integron-positive Salmonella strains belonging to serotypes Infantis, Enteritidis, and Typhimurium were attributed to two, three, and five different pulsotypes, respectively. The findings indicated that the distribution and drug resistance pattern of most prevalent Salmonella serotypes were broadly similar to that reported globally from human isolates. Presence of class 1 integrons was common among Salmonella serotypes in Tehran, Iran. Concurrent clonal expansion and horizontal transmission events seem to contribute to increase in drug resistance prevalence among Salmonella serotypes.     

耐碳青霉烯酶类肺炎克雷伯菌中整合子及相关基因盒的分布

目的 了解耐碳青霉烯类肺炎克雷伯菌整合子及相关基因盒的分布,分析整合子与耐碳青霉烯类肺炎克雷伯菌多药耐药的关系.方法 采用革兰阴性杆菌药敏卡检测耐碳青霉烯类肺炎克雷伯菌对抗菌药物的敏感性,聚合酶链反应(PCR)法检测Ⅰ、Ⅱ、Ⅲ类整合酶基因(intⅠ1、intⅡ2、intⅢ3)及Ⅰ类整合子可变医基因盒,并分析可变区上游启动子的类型.结果 74株耐碳青霉烯类肺炎克雷伯菌中有67株检测到Ⅰ类整合酶基因,检出率为90.5％,2株检测到Ⅱ类整合酶基因,检出率为2.7％,未检测到Ⅲ类整合酶基因阳性菌株;55株(74.3％)成功扩增出Ⅰ类整合子可变区,主要携带甲氧苄啶及早期使用的氨基糖苷类抗菌药物耐药基因,可变区启动子大多为弱启动子;携带Ⅰ类整合子菌株对常用抗菌药物的耐药率与Ⅰ类整合子阴性菌株之间的差异无统计学意义.结论 Ⅰ类整合子及相关基因盒在耐碳青霉烯类肺炎克雷伯菌中分布广泛.所携带的整合子具有非常强的从周围环境中捕获耐药性基因盒的能力.