integron

整合子
  • 文章类型: Journal Article
    这项研究探讨了结直肠癌(CRC)患者中粘附性侵袭性大肠杆菌(AIEC)的患病率,并研究了有效的细胞内抗生素作为AIEC感染CRC患者治疗策略的潜力。考虑到整合子在细菌抗生素耐药性中的关键作用,从CRC患者中分离出的AIEC中1类和2类整合子的频率,在伊斯法罕的3个胃肠病诊所之一,伊朗被检查。从结直肠活检中分离出AIEC菌株,并使用圆盘扩散法评估其抗微生物敏感性。采用聚合酶链反应(PCR)检测intl1和intl2。使用多位点序列分型(MLST)方法对10个选定的分离株进行分型。在150个样本中,24被确定为AIEC,从CRC2(33.4%)和CRC1(29.16%)中分离出的数量最高,FH组(8.3%)和对照组(12.5%)最少。发现79.2%的AIEC菌株中的int1和45.8%的int2,并且41.6%的菌株具有两个整合子。具有int1的AIEC分离株对甲氧苄啶-磺胺甲恶唑的敏感性最高(57.9%),而int2对环丙沙星的敏感性最高(63.6%)。在AIEC分离株中观察到对利福平的抗性与整合子2的存在之间存在显着关联。此外,整合子1存在之间的显著相关性,入侵,生存,并鉴定了AIEC菌株中巨噬细胞内的复制。MLST分析显示来自CC131的ST131,整合子1是最常见的序列类型(ST)。此类菌株在CRC人群中的出现构成了严重的公共卫生威胁。STS的分布格局在研究群体之间有所不同,大流行的STs强调了检查和治疗感染这些分离株的患者的重要性。有必要进行全面的前瞻性临床研究,以评估在CRC中检测这种病理改变的预后价值,并评估针对耐药AIEC的治疗技术。比如噬菌体疗法,细菌素,和抗粘连化合物,用于CRC的预防和治疗。
    This study explores the prevalence of adherent-invasive Escherichia coli (AIEC) in colorectal cancer (CRC) patients and investigates the potential of effective intracellular antibiotics as a therapeutic strategy for CRC patients with AIEC infections. Considering the pivotal role of integrons in bacterial antibiotic resistance, the frequency of class 1 and 2 integrons in AIEC isolated from CRC patients, in one of the referenced 3 gastroenterology clinics in Isfahan, Iran was examined. AIEC strains were isolated from the colorectal biopsies and their antimicrobial sensitivity was assessed using the disc diffusion method. Polymerase chain reaction (PCR) was employed to detect intl1 and intl2. The multilocus sequence typing (MLST) method was utilized to type 10 selected isolates. Of the 150 samples, 24 were identified as AIEC, with the highest number isolated from CRC2 (33.4%) and CRC1 (29.16%), and the least from the FH group (8.3%) and control group (12.5%). int1 in 79.2% and int2 in 45.8% of AIEC strains were found and 41.6% of strains had both integrons. AIEC isolates with int1 exhibited the highest sensitivity to trimethoprim-sulfamethoxazole (57.9%), while those with int2 showed the highest sensitivity to ciprofloxacin (63.6%). A significant association between resistance to rifampin and integron 2 presence in AIEC isolates was observed. Furthermore, a significant correlation between integron 1 presence, invasion, survival, and replication within macrophages in AIEC strains was identified. MLST analysis revealed ST131 from CC131 with integron 1 as the most common sequence type (ST). The emergence of such strains in CRC populations poses a serious public health threat. The distribution pattern of STs varied among studied groups, with pandemic STs highlighting the importance of examining and treating patients infected with these isolates. Comprehensive prospective clinical investigations are warranted to assess the prognostic value of detecting this pathovar in CRC and to evaluate therapeutic techniques targeting drug-resistant AIECs, such as phage therapy, bacteriocins, and anti-adhesion compounds, for CRC prevention and treatment.
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  • 文章类型: Journal Article
    铜绿假单胞菌作为机会病原体产生几种毒力因子。这项研究评估了外切酶(exo)A的相对频率,U和S基因和整合子类(I,II,和III)在阿瓦兹烧伤患者的多药耐药临床铜绿假单胞菌中,伊朗西南部。
    在该横断面研究中,从355个伤口样品中回收铜绿假单胞菌分离物。根据临床和实验室标准研究所,通过圆盘琼脂扩散法在Muller-Hinton琼脂上进行抗菌敏感性试验。如果MDR分离株同时对3种抗生素表现出耐药性,则将其定义。广泛耐药被定义为对除两个或更少的抗菌类别中的至少一种药物不敏感。一班的存在,II,和III整合子和毒力基因使用PCR测定法对提取的DNA进行测定。
    总的来说,145个临床铜绿假单胞菌分离物用生化和PCR试验证实。总的来说,35%(52/145)的分离株来自男性,64%(93/145)来自女性住院烧伤患者。铜绿假单胞菌对抗生素耐药率最高的是哌拉西林59%(n=86/145)和哌拉西林他唑巴坦57%(n=83/145)。总共100%的分离株对至少一种抗生素具有抗性。MDR和XDR铜绿假单胞菌的频率为60%和29%,分别。整合子I类的患病率,II,铜绿假单胞菌中的III为60%,7.58%,和3.44%,分别。IntI在MDR和XDR铜绿假单胞菌中更常见。此外,70(48%)的铜绿假单胞菌分离株没有整合子基因。此外,exoA,exoS,铜绿假单胞菌中的exoU频率为55%,55%,56%,分别。
    发现铜绿假单胞菌作为一种有效的病原体,在卫生界具有强毒力因子和高抗生素耐药性,可引起烧伤患者的难治性疾病。
    UNASSIGNED: Pseudomonas aeruginosa as an opportunistic pathogen produces several virulence factors. This study evaluated the relative frequency of exoenzymes (exo) A, U and S genes and integron classes (I, II, and III) among multi-drug-resistant clinical P. aeruginosa isolates from burn patients in Ahvaz, southwest of Iran.
    UNASSIGNED: In this cross-sectional study P. aeruginosa isolates were recovered from 355 wound samples. The antimicrobial susceptibility test was done by disk agar diffusion method on Muller-Hinton agar according to the Clinical and Laboratory Standards Institute. MDR isolates were defined if they showed simultaneous resistance to 3 antibiotics. Extensively drug-resistant was defined as nonsusceptibility to at least one agent in all but two or fewer antimicrobial categories. The presence of class I, II, and III integrons and virulence genes was determined using a PCR assay on extracted DNA.
    UNASSIGNED: Overall, 145 clinical P. aeruginosa isolates were confirmed with biochemical and PCR tests. Overall, 35% (52/145) of the isolates were taken from males and 64% (93/145) from female hospitalized burn patients. The highest resistance rates of P. aeruginosa isolates to antibiotics were related to piperacillin 59% (n = 86/145) and piperacillin-tazobactam 57% (n = 83/145). A total of 100% of isolates were resistant to at least one antibiotic. MDR and XDR P. aeruginosa had a frequency of 60% and 29%, respectively. The prevalence of integron classes I, II, and III in P. aeruginosa was 60%, 7.58%, and 3.44%, respectively. IntI was more common in MDR and XDR P. aeruginosa isolates. In addition, 70(48%) of P. aeruginosa isolates did not harbor integron genes. Besides, exoA, exoS, and exoU in P. aeruginosa had a frequency of 55%, 55%, and 56%, respectively.
    UNASSIGNED: It was found that P. aeruginosa as a potent pathogen with strong virulence factors and high antibiotic resistance in the health community can cause refractory diseases in burn patients.
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  • 文章类型: Journal Article
    UNASSIGNED: The aims of this study were to: (i) determine antibiotic susceptibility of clinical Stenotrophomonas maltophilia isolates, (ii) investigate the presence of different classes of integrons and sul genes responsible for sulphonamide resistance, (iii) assess the molecular epidemiology of the isolates by determining their clonal relatedness, and (iv) investigate the potential sources of infection by collecting environmental samples when necessary.
    UNASSIGNED: 99 S. maltophilia isolates from clinical specimens of hospitalized patients were screened by PCR for sul1, sul2, sul3 genes, and integron-associated integrase genes: intI1, intI2, and intI3. PFGE was used to determine the clonal relatedness of the isolates.
    UNASSIGNED: Susceptibility rates for trimethoprim-sulfamethoxazole, levofloxacin, and ceftazidime were 90.9%, 91.9%, and 53.5% respectively. All trimethoprim-sulfamethoxazole-resistant isolates were positive for intI1 and sul1. PFGE analysis revealed that 24 of the isolates were clonally related, clustering in seven different clones. Five of the nine trimethoprim-sulfamethoxazole-resistant isolates were clonally related. The first isolate in this clone was from a wound sample of a patient in the infectious diseases clinic, and the other four were isolated from the bronchoalveolar lavage samples of patients in the thoracic surgery unit. The patient with the first isolate neither underwent bronchoscopy nor stayed in the thoracic surgery unit. Although clustering was observed in bronchoalveolar lavage samples, no S. maltophilia growth was detected in environmental samples.
    UNASSIGNED: The findings demonstrated that the sul1 gene carried by class 1 integrons plays an important role in trimethoprim-sulfamethoxazole resistance in S. maltophilia isolates. PFGE analysis revealed a high degree of genetic diversity. However, detection of clonally related isolates suggests the acquisition from a common source and/or cross-transmission of this microorganism between the patients.
    UNASSIGNED: In dieser Studie sollten (i) die Antibiotika-Empfindlichkeit klinischer Stenotrophomonas maltophilia-Isolate bestimmt, (ii) das Vorhandensein verschiedener, für die Sulfonamid-Resistenz verantwortlichen Klassen von Integronen und sul-Genen, untersucht, iii) die molekulare Epidemiologie der Isolate durch Bestimmung ihrer klonalen Verwandtschaft bewertet und (iv) potenzielle Infektionsquellen durch Umgebungsuntersuchungen überprüft werden.
    UNASSIGNED: 99 S. maltophilia-Isolate aus klinischen Proben von Krankenhauspatienten wurden mittels PCR auf die Gene sul1, sul2 und sul3 sowie die Integron-assoziierten Integrase-Gene intI1, intI2 und intI3 untersucht. Mit PFGE wurde die klonale Verwandtschaft der Isolate bestimmt.
    UNASSIGNED: Die Empfindlichkeitsraten für Trimethoprim-Sulfamethoxazol, Levofloxacin und Ceftazidim betrugen 90,9%, 91,9% bzw. 53,5%. Alle Trimethoprim-Sulfamethoxazol-resistenten Isolate waren positiv für intI1 und sul1. Die PFGE-Analyse ergab, dass 24 der Isolate klonal verwandt waren und sich in sieben verschiedenen Klonen zusammenschlossen. Fünf der neun Trimethoprim-Sulfamethoxazol-resistenten Isolate waren klonal verwandt. Das erste Isolat dieses Klons stammte aus der Wunde eines Patienten in der Klinik für Infektionskrankheiten, die anderen vier wurden aus bronchoalveolären Lavageproben von Patienten in der Thoraxchirurgie isoliert. Der Patient mit dem ersten Isolat unterzog sich weder einer Bronchoskopie noch hielt er sich in der Abteilung für Thoraxchirurgie auf. Obwohl in den bronchoalveolären Lavageproben eine Clusterbildung beobachtet wurde, war S. maltophilia nicht in den Umgebungsuntersuchungen nachweisbar.
    UNASSIGNED: Die Ergebnisse zeigen, dass das von Klasse-1-Integronen getragene sul1-Gen eine wichtige Rolle bei der Trimethoprim-Sulfamethoxazol-Resistenz von S. maltophilia-Isolaten spielt. Die PFGE-Analyse ergab ein hohes Maß an genetischer Vielfalt. Der Nachweis klonal verwandter Isolate deutet jedoch darauf hin, dass dieser Mikroorganismus aus einer gemeinsamen Quelle stammt und/oder zwischen den Patienten übertragen wird.
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  • 文章类型: Journal Article
    目的:研究福氏志贺氏菌对磺胺甲恶唑-甲氧苄啶(SXT)的耐药性之间的相关性(S。flexneri)以及整合子和相关抗生素抗性基因的存在。
    方法:收集2012-2020年济南市腹泻患儿粪便中分离的福氏志贺菌115株,采用Etest法测定SXT的最低抑菌浓度(MIC)。1类,2类和3类整合子基因的存在,可变区抗生素抗性基因盒,使用聚合酶链反应(PCR)检测sul1,sul2,sul3和SXT元件。通过DNA测序和BLAST比较进一步分析阳性结果。
    结果:总计,115株福氏链球菌对SXT的耐药率为60.9%。1类和2类整合子的患病率分别为88.7%和87.0%,分别,没有检测到3类整合子。在菌株中,13.0%携带具有可变区抗生素抗性基因盒dfrA17-aadA5和dfrV的典型1类整合子,而85.2%携带具有可变区抗生素抗性基因盒blaoxa-30-aadA1的非典型1类整合子。2类整合子的可变区抗生素抗性基因盒均为dfrA1sat1aadA1。SXT敏感型和耐药型福氏链球菌1类整合子和2类整合子的存在差异有统计学意义(χ2=22.800,χ2=16.365,P<0.01,P<0.01)。通过整合子携带dfrV和dfrA1的整合子在SXT抗性上也显示出统计学差异(χ2=9.422,χ2=16.365,P<0.01,P<0.01)。PCR显示在13.0%和47.0%的菌株中存在sul1和sul2,分别,没有检测到sul3和SXT元素。sul1、sul2的存在在SXT敏感型和耐药型福氏杆菌菌株之间存在显著差异(χ2=9.588,χ2=65.445,P<0.01,P<0.01)。
    结论:总之,整合子参与了福氏杆菌的SXT抗性,和dfrV,dfrA1,sul1,sul2与福氏杆菌的SXT抗性密切相关。
    OBJECTIVE: To investigate the correlation between sulfamethoxazole-trimethoprim (SXT) resistance in Shigella flexneri (S.flexneri) and the presence of integrons and relevant antibiotic resistance genes.
    METHODS: We collected 115 strains of Shigella flexneri isolated from feces of children with diarrhea in Jinan from 2012 to 2020 and determined the minimum inhibitory concentration (MIC) of SXT by Etest method. The presence of class 1, class 2, and class 3 integron genes, variable region antibiotic resistance gene cassettes, and sul1, sul2, sul3, and SXT elements were detected using polymerase chain reaction (PCR). Positive results were further analyzed by DNA sequencing and BLAST comparison.
    RESULTS: In total, the resistance rate to SXT was 60.9% among the 115 S.flexneri strains. The prevalence of class 1 and class 2 integrons were 88.7% and 87.0%, respectively, with no class 3 integrons detected. Among the strains, 13.0% carried typical class 1 integrons with variable region antibiotic resistance gene cassettes dfrA17-aadA5 and dfrV, while 85.2% carried atypical class 1 integrons with variable region antibiotic resistance gene cassette blaoxa-30-aadA1. The variable region antibiotic resistance gene cassettes of class 2 integrons were all dfrA1+sat1+aadA1. There was a statistical difference between the presence of class 1 integrons and class 2 integrons between the SXT-sensitive and resistant S.flexneri strains (χ2=22.800, χ2=16.365, P<0.01, P<0.01). Integrons carrying dfrV and dfrA1 by integrons also showed a statistical difference in SXT resistance (χ2=9.422, χ2=16.365, P<0.01, P<0.01). PCR revealed the presence of sul1 and sul2 in 13.0% and 47.0% of strains, respectively, with neither sul3 nor SXT elements detected. There was a significant difference between the presence of sul1, sul2 between the SXT-sensitive and resistant S.flexneri strains (χ2=9.588, χ2=65.445, P<0.01, P<0.01).
    CONCLUSIONS: In summary, integrons are involved in SXT resistance of S.flexneri, and dfrV, dfrA1, sul1, sul2 are closely related to SXT resistance of S.flexneri.
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  • 文章类型: Journal Article
    鲍曼不动杆菌是一种非发酵革兰阴性菌,可引起危重病患者医院感染。耐碳青霉烯类鲍曼不动杆菌(CRAB)在临床上迅速传播,并已成为一个关键问题。这项研究的主要目的是确定CRAB分离株中整合子和生物膜形成相关毒力基因的分布。总共收集了269个鲍曼不动杆菌分离株(219个CRAB分离株和50个碳青霉烯敏感鲍曼不动杆菌(CSAB)分离株)。碳青霉烯酶基因(blaKPC,BlaVIM,blaIMP,blaNDM,和blaOXA-23样)和生物膜形成相关的毒力基因(abal,bfms,bap,和cusE)用PCR筛选。用PCR筛选1类整合子,和常见的启动子和基因盒阵列通过限制性模式分析结合引物步行测序确定。进行了全基因组测序,并分析了BlaOXA-23样阴性分离株的数据。所有219个CRAB分离株均为blaKPC阴性,BlaVIM,blaIMP,和BLANDM,而在218个分离株中检测到blaOXA-23样。abal的检出率,bfms,bap,219名CRAB中的cusE为93.15%,63.93%,88.13%,和77.63%,分别。在75个CRAB(34.25%)和3个CSAB中检测到1类整合子。在1类整合子中检测到具有相对强的PcH2启动子的单基因盒阵列aacA4-catB8-aadA1。发现blaOXA-23样阴性CRAB分离株是携带blaOXA-72,blaOXA-259和blaADC-26的新序列类型(牛津3272,巴斯德2520)。总之,BLAOXA-23样是CRAB对碳青霉烯类耐药的主要原因。据报道,一种新的(牛津3272,巴斯德2520)CRAB序列类型携带blaOXA-72,blaOXA-259和blaADC-26。
    Acinetobacter baumannii is a non-fermentative Gram-negative bacterium that can cause nosocomial infections in critically ill patients. Carbapenem-resistant A. baumannii (CRAB) has spread rapidly in clinical settings and has become a key concern. The main objective of this study was to identify the distribution of integrons and biofilm-formation-related virulence genes in CRAB isolates. A total of 269 A. baumannii isolates (219 isolates of CRAB and 50 isolates of carbapenem-sensitive A. baumannii (CSAB)) were collected. Carbapenemase genes (bla KPC, bla VIM, bla IMP, bla NDM, and bla OXA-23-like) and biofilm-formation-related virulence genes (abal, bfms, bap, and cusE) were screened with PCR. Class 1 integron was screened with PCR, and common promoters and gene cassette arrays were determined with restriction pattern analysis combined with primer walking sequencing. Whole-genome sequencing was conducted, and data were analyzed for a bla OXA-23-like-negative isolate. All 219 CRAB isolates were negative for bla KPC, bla VIM, bla IMP, and bla NDM, while bla OXA-23-like was detected in 218 isolates. The detection rates for abal, bfms, bap, and cusE in 219 CRAB were 93.15%, 63.93%, 88.13%, and 77.63%, respectively. Class 1 integron was detected in 75 CRAB (34.25%) and in 3 CSAB. The single gene cassette array aacA4-catB8-aadA1 with relatively strong PcH2 promoter was detected in class 1 integrons. The bla OXA-23-like-negative CRAB isolate was revealed to be a new sequence type (Oxford 3272, Pasteur 2520) carrying bla OXA-72, bla OXA-259, and bla ADC-26. In conclusion, bla OXA-23-like was the main reason for CRAB\'s resistance to carbapenems. A new (Oxford 3272, Pasteur 2520) CRAB sequence type carrying the bla OXA-72, bla OXA-259, and bla ADC-26 was reported.
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  • 文章类型: Case Reports
    耐碳青霉烯类沙门氏菌(S.enterica)对公共卫生构成重大威胁,引起胃肠炎和侵袭性感染.我们报道了耐碳青霉烯的肠链球菌血清London菌株的首次出现,A132,在中国携带blaNDM-5基因。全基因组测序和生物信息学分析将A132指定为ST155,这是中国经常报道的多药耐药克隆。菌株A132表现出对多种抗生素的抗性,确定了20个获得性抗生素抗性基因(ARGs),主要位于IncFIB质粒(pA132-1-NDM)上。值得注意的是,blaNDM-5基因位于IS26侧翼的1类整合子-ISR1复合物中,包含两个基因盒。一个卡带是1类整合子,这可能有助于整个建筑群的传输,而另一个是含有blaNDM-5的ISR1-IS26侧翼盒,携带多个其他ARG。基于携带blaNDM-5的盒的Genbank数据库搜索确定了在大肠杆菌(p91)和ormaechei肠杆菌(p388)的可传播的IncFIA质粒中发现的相似遗传背景,具有共享的宿主范围,提示blaNDM-5跨物种传播的可能性。据我们所知,这是首例报道的携带blaNDM-5基因的沙门氏菌伦敦ST155。系统发育分析表明,从同一省分离的A132和8个伦敦ST155菌株之间存在密切的关系。然而,A132的不同之处在于携带blaNDM-5基因和四个独特的ARG。鉴于携带blaNDM-5和18种其他ARG的F型质粒的高传播性,必须实施警惕的监测和采取适当的感染控制措施,以减轻对公众健康的威胁。
    Carbapenem-resistant Salmonella enterica (S. enterica) pose a significant threat to public health, causing gastroenteritis and invasive infections. We report the first emergence of a carbapenem-resistant S. enterica serovar London strain, A132, carrying the blaNDM-5 gene in China. Whole-genome sequencing and bioinformatics analysis assigned A132 to be ST155, a multidrug-resistant clone frequently reported in China. The strain A132 exhibited resistance to multiple antibiotics, with 20 acquired antibiotic resistance genes (ARGs) identified, predominantly located on the IncFIB plasmid (pA132-1-NDM). Notably, the blaNDM-5 gene was located within an IS26 flanked-class 1 integron-ISCR1 complex, comprising two genetic cassettes. One cassette is the class 1 integron, which may facilitate the transmission of the entire complex, while the other is the blaNDM-5-containing ISCR1-IS26-flanked cassette, carrying multiple other ARGs. Genbank database search based on the blaNDM-5-carrying cassette identified a similar genetic context found in transmissible IncFIA plasmids from Escherichia coli (p91) and Enterobacter hormaechei (p388) with a shared host range, suggesting the potential for cross-species transmission of blaNDM-5. To our knowledge, this is the first reported case of Salmonella serovar London ST155 harboring blaNDM-5 gene. Phylogenetic analysis indicated a close relationship between A132 and eight S. London ST155 strains isolated from the same province. However, A132 differed by carrying the blaNDM-5 gene and four unique ARGs. Given the high transmissibility of the F-type plasmid harboring blaNDM-5 and 18 other ARGs, it is imperative to implement vigilant surveillance and adopt appropriate infection control measures to mitigate the threat to public health.
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  • 文章类型: Journal Article
    肺炎克雷伯菌(hvKp)的“高毒力”变种是一种新兴的病原体,可引起危及生命的感染。进行本研究以确定hvKp的患病率并调查这些分离物中1、2和3类整合子的存在。
    在三家教学医院进行了一项横断面研究,Ahvaz,伊朗西南部,从2019年1月1日至2020年12月31日。从住院患者收集样品,并且仅包括从每个患者收集的第一样品。使用生化测试从不同标本中分离肺炎克雷伯菌菌株,并通过靶向16S-23SrDNA内部转录间隔区进行确认。使用字符串测试回收HvKp分离株,并通过检测毒力相关基因(rmpA,iuca,和magA)。使用圆盘扩散法确定分离株的抗生素敏感性模式。筛选分离株是否存在整合子基因(intI,intII,和intIII)和基于重复元件序列的聚合酶链反应(PCR)来确定菌株相关性。采用SPSS版本22进行数据分析。
    七十一(77%)的分离株显示多药耐药(MDR)表型。HvKP占血液分离的cKp(46%)和尿路感染(38%)的14%(13/92),其中绝大多数(61.5%;8/13)表现为MDR表型。使用PCR检测,发现92个分离株中的29个(31.5%)对IntI的存在具有阳性结果。IntI阳性菌株中的三个是hvKP。2类整合子存在于8/92cKp分离株中。在3/8个分离株中发现2类整合子与1类整合子共存。所有整合子阳性分离株(IntI和/或IntII)对至少三种不同类型的抗生素具有抗性,并显示MDR表型。在分离物中没有检测到3类整合子。
    我们的研究结果表明,考虑到整合子在促进细菌中抗性基因的获取和传播中的作用,监测hvKp的出现,强调抗菌素耐药性的机制,可以防止产生碳青霉烯酶的hvKp菌株的传播。
    UNASSIGNED: The \"hypervirulent\" variant of Klebsiella pneumoniae (hvKp) is an emerging pathogen that cause life-threatening infection. The present study was conducted to identify the prevalence of hvKp and to investigate the presence class 1, 2, and 3 integrons in these isolates.
    UNASSIGNED: A cross-sectional study was conducted at three teaching hospitals, Ahvaz, South-west of Iran, from January 1, 2019 to December 31, 2020. Samples were collected from inpatients and included only the first samples collected from each patient. K. pneumoniae strains were isolated from different specimens using biochemical test and confirmed by targeting 16S-23S rDNA internal transcribed spacer. HvKp isolates were recovered using string test and were further characterized by detection virulence-associated genes (rmpA, iucA, and magA). Antibiotic susceptibility patterns of isolates were determined using the disc diffusion method. Isolates were screened for presence the integron genes (intI, intII, and intIII) and repetitive element sequence-based polymerase chain reaction (PCR) performed to determine strain relatedness. SPSS version 22 was used for the data analysis.
    UNASSIGNED: Seventy-one (77%) of isolates showed multidrug-resistant (MDR) phenotype. HvKP accounted for 14% (13/92) of cKp isolated from blood (46%) and urinary tract infection (38%), and the great majority of them (61.5%; 8/13) exhibited MDR phenotype. Using the PCR assay, 29 of 92 isolates (31.5%) were found to have positive results for the presence of IntI. Three of the IntI-positive strains were hvKP. Class 2 integron was present in 8/92 cKp isolates. Integron Class 2 was found to coexist with Class 1 integron in 3/8 isolates. All integron-positive isolates (IntI and/or IntII) were resistant to at least three different classes of antibiotics and showed MDR phenotype. No Class 3 integrons were detected among the isolates.
    UNASSIGNED: The results of our study revealed that considering the role of integrons in facilitating the acquisition and dissemination of resistance genes among bacteria, monitoring the emergence of hvKp, emphasizing on the mechanism of antimicrobial resistance, can prevent from the spread of carbapenemase-producing hvKp strains.
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  • 文章类型: Journal Article
    有关日本使用碳青霉烯酶的产气克雷伯菌的信息有限。2014年9月至2022年12月进行了一项全国性的全面调查,从日本的57个医疗机构中分离出67种非重复的耐碳青霉烯K.aerogenes菌株。通过基因检测和全基因组测序,发现六株菌株具有碳青霉烯酶,包括亚胺酶(IMP)-1,IMP-6,新德里金属-β-内酰胺酶(NDM)-1和NDM-5。携带blaNDM-5的菌株是新菌株ST709,属于中国主要的ST4的克隆复合物。含有blaIMP-1的新型整合子具有恶草素酶-101基因,这是一个以前未报告的结构,具有IncN4质粒类型。然而,在具有blaIMP-6的菌株中发现的整合子,这是最常见的鉴定,与国内报道的肺炎克雷伯菌相匹配,表明相同的整合子的流行。含有blaNDM的转座子与先前在日本报道的携带blaNDM-5的产气K.表明相同类型的转座子可能已经传播到日本的产气K.这项调查分析了移动遗传元素,如整合子和转座子,为了了解碳青霉烯酶的传播,强调了日本耐碳青霉烯类肠杆菌日益增长的挑战,并强调了持续监测以控制这些病原体的迫切需要。
    Information regarding Klebsiella aerogenes haboring carbapenemase in Japan is limited. A comprehensive nationwide survey was conducted from September 2014 to December 2022, and 67 non-duplicate strains of carbapenem-resistant K. aerogenes were isolated from 57 healthcare facilities in Japan. Through genetic testing and whole-genome sequencing, six strains were found to possess carbapenemases, including imipenemase (IMP)-1, IMP-6, New Delhi metallo-β-lactamase (NDM)-1, and NDM-5. The strain harboring blaNDM-5 was the novel strain ST709, which belongs to the clonal complex of the predominant ST4 in China. The novel integron containing blaIMP-1 featured the oxacillinase-101 gene, which is a previously unreported structure, with an IncN4 plasmid type. However, integrons found in the strains possessing blaIMP-6, which were the most commonly identified, matched those reported domestically in Klebsiella pneumoniae, suggesting the prevalence of identical integrons. Transposons containing blaNDM are similar or identical to the transposon structure of K. aerogenes harboring blaNDM-5 previously reported in Japan, suggesting that the same type of transposon could have been transmitted to K. aerogenes in Japan. This investigation analyzed mobile genetic elements, such as integrons and transposons, to understand the spread of carbapenemases, highlighting the growing challenge of carbapenem-resistant Enterobacterales in Japan and underscoring the critical need for ongoing surveillance to control these pathogens.
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  • 文章类型: Journal Article
    碳青霉烯酶基因blaIMP整合到碳青霉烯耐药鲍曼不动杆菌(CRAB)的染色体中尚未见报道。这项研究的目的是探索从2011年诊断为菌血症的台湾患者中分离出的CRABAB322的基因组特征,该患者的染色体带有blaIMP-19。采用圆盘扩散和肉汤微量稀释来分析AB322对14种抗菌剂的抗菌敏感性。利用纳米孔全基因组测序平台进行AB322基因组测序,并进一步进行结合以研究blaIMP-19对阿米卡星耐药的鲍曼不动杆菌218(AB218)和医院不动杆菌254(AN254)的转移性。结果表明,AB322被归类为多重耐药鲍曼不动杆菌,但对氨苄西林/舒巴坦仍然敏感。粘菌素,还有替加环素.全基因组测序揭示了AB322基因组,由4,098,985bp的染色体组成,一个名为pAB322-1的71,590-bp接合质粒和一个名为pAB322-2的8,726-bp质粒。多位点序列分型分析表明,AB322属于1型序列。AB322染色体包含许多获得性抗菌素抗性基因,包括aph(3')-Ia,aadA1b,aadA1,aac(6')-Ib3,aac(3)-Ia,blaADC-25、blaOXA-69、blaIMP-19、catA1、sul1和tet(A),赋予β-内酰胺抗性,氨基糖苷类,氯霉素,磺胺甲恶唑,还有四环素.此外,blaIMP-19被鉴定为位于AB322染色体上的1类整合子In240和不完整的PHAGE_Salmon_SJ46_NC_031129内。然而,结合实验表明,在我们的测试条件下,blaIMP-19不能转移到AB218和AN254中。总之,我们首先报道了CRAB中染色体整合的blaIMP-19的存在,可能由整合子介导。blaIMP-19在不同物种中的未来传播,导致碳青霉烯耐药性传播,需要密切监测。
    目的:抗菌素耐药基因的水平转移对于耐药性的传播至关重要,特别是鲍曼不动杆菌已成为临床上重要的病原体。然而,在这项研究中,我们首先报道了blaIMP-19基因整合到鲍曼不动杆菌的染色体中,这种水平转移可能与整合子-噬菌体元件有关。此外,这些携带抗微生物药物抗性基因的DNA片段可能通过水平移动到接合质粒上而进一步传播到其他病原体。
    Integration of carbapenemase gene blaIMP into the chromosome of carbapenem-resistant Acinetobacter baumannii (CRAB) has not been reported. The aim of this study was to explore the genomic characteristics of CRAB AB322 isolated from a Taiwanese patient diagnosed with bacteremia in 2011, whose chromosome harbors blaIMP-19. Disk diffusion and broth microdilution were employed to analyze the antimicrobial susceptibility of AB322 to 14 antimicrobials. Nanopore whole-genome sequencing platform was utilized for AB322 genome sequencing, and conjugation was further performed to investigate the transferability of blaIMP-19 to amikacin-resistant A. baumannii 218 (AB218) and Acinetobacter nosocomialis 254 (AN254). The results showed that AB322 was classified as multidrug-resistant A. baumannii but remained susceptible to ampicillin/sulbactam, colistin, and tigecycline. Whole-genome sequencing revealed the AB322 genome, consisting of a 4,098,985-bp chromosome, a 71,590-bp conjugative plasmid named pAB322-1, and an 8,726-bp plasmid named pAB322-2. Multilocus sequence typing analysis indicated that AB322 belonged to sequence type 1. AB322 chromosome harbored numerous acquired antimicrobial resistance genes, including aph(3\')-Ia, aadA1b, aadA1, aac(6\')-Ib3, aac (3)-Ia, blaADC-25, blaOXA-69, blaIMP-19, catA1, sul1, and tet(A), conferring resistance to β-lactams, aminoglycosides, chloramphenicol, sulfamethoxazole, and tetracyclines. Moreover, blaIMP-19 was identified to be situated within class 1 integron In240 and an incomplete PHAGE_Salmon_SJ46_NC_031129 on AB322 chromosome. However, conjugation experiments revealed that blaIMP-19 could not be transferred to AB218 and AN254 in our testing conditions. In conclusion, we first report the presence of chromosomal-integrated blaIMP-19 in CRAB, possibly mediated by integron. The future dissemination of blaIMP-19 among different species, leading to carbapenem resistance dissemination, requires close monitoring.
    OBJECTIVE: The horizontal transfer of antimicrobial-resistant genes is crucial for the dissemination of resistance, especially as Acinetobacter baumannii has emerged as a clinically significant pathogen. However, in this study, we first report the integration of the blaIMP-19 gene into the chromosome of A. baumannii, and such horizontal transfer may be associated with integron-phage elements. Additionally, it is possible that these DNA fragments carrying antimicrobial-resistant genes could further spread to other pathogens by moving horizontally onto conjugative plasmids.
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  • 文章类型: Journal Article
    日本是碳青霉烯类耐药铜绿假单胞菌(CRPA)患病率约10%的国家。目前,缺乏对日本CRPA基因型和表型模式的全面概述.在这里,我们对2019年至2020年从日本78家医院收集的382株美罗培南耐药CRPA分离株进行了基因组测序和定量药敏试验.CRPA的敏感性为52.9%,26.4%,88.0%对抗哌拉西林他唑巴坦,环丙沙星,和阿米卡星,分别,而27.7%的CRPA分离株被归类为难以治疗的耐药铜绿假单胞菌。在检测到的148种序列类型中,ST274(9.7%)占主导地位,其次是ST235(7.6%)。ST235中尿液分离株的比例高于其他STs(P=0.0056,χ2检验)。只有4.1%的CRPA分离物携带碳青霉烯酶基因:blaGES(2)和blaIMP(13)。一种ST235分离物在染色体中携带新的blaIMP变体blaIMP-98。关于染色体突变,87.1%的CRPA分离株在oprD中具有失活或其他抗性突变,和28.8%显示突变的调控基因(mexR,nalC,和nalD)用于MexAB-OprM外排泵。此外,4.7%的CRPA分离株在PBP3编码基因ftsI中携带抗性突变。这项研究和其他监测研究的结果共同表明,CRPA表现出明显的遗传多样性,其在日本的多药耐药性不如其他地区流行。这项研究提供了一个有价值的数据集,解决了亚太地区关于CRPA的基因型/表型信息的差距,地区之间的流行病学背景明显不同。
    Japan is a country with an approximate 10% prevalence rate of carbapenem-resistant Pseudomonas aeruginosa (CRPA). Currently, a comprehensive overview of the genotype and phenotype patterns of CRPA in Japan is lacking. Herein, we conducted genome sequencing and quantitative antimicrobial susceptibility testing for 382 meropenem-resistant CRPA isolates that were collected from 78 hospitals across Japan from 2019 to 2020. CRPA exhibited susceptibility rates of 52.9%, 26.4%, and 88.0% against piperacillin-tazobactam, ciprofloxacin, and amikacin, respectively, whereas 27.7% of CRPA isolates was classified as difficult-to-treat resistance P. aeruginosa. Of the 148 sequence types detected, ST274 (9.7%) was predominant, followed by ST235 (7.6%). The proportion of urine isolates in ST235 was higher than that in other STs (P = 0.0056, χ2 test). Only 4.1% of CRPA isolates carried the carbapenemase genes: blaGES (2) and blaIMP (13). One ST235 isolate carried the novel blaIMP variant blaIMP-98 in the chromosome. Regarding chromosomal mutations, 87.1% of CRPA isolates possessed inactivating or other resistance mutations in oprD, and 28.8% showed mutations in the regulatory genes (mexR, nalC, and nalD) for the MexAB-OprM efflux pump. Additionally, 4.7% of CRPA isolates carried a resistance mutation in the PBP3-encoding gene ftsI. The findings from this study and other surveillance studies collectively demonstrate that CRPA exhibits marked genetic diversity and that its multidrug resistance in Japan is less prevailed than in other regions. This study contributes a valuable data set that addresses a gap in genotype/phenotype information regarding CRPA in the Asia-Pacific region, where the epidemiological background markedly differs between regions.
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