molecular typing

分子分型
  • 文章类型: Journal Article
    目的:为了研究抗菌药的遗传特征和耐药性,包括主要的β-内酰胺抗生素抗性基因,在累西腓-PE三级医院分离的鲍曼不动杆菌中,巴西,在后COVID-19大流行期间。
    结果:A.鲍曼不动杆菌分离株在2023年至2024年间从不同的临床样本中收集。抗菌素耐药性测试遵循标准化方案,通过PCR和测序检测β-内酰胺酶编码基因。还对blaOXA-碳青霉烯酶和blaADC基因上游的ISAba1进行了调查。通过ERIC-PCR评估遗传多样性。在78个鲍曼不动杆菌中,对多种抗菌药物的广泛耐药是显而易见的。各种获得的β-内酰胺酶编码基因(blaOXA-23,-24,-58,-143,blaVIM,和blaNDM)被检测到。此外,这是blaVIM-2在巴西携带blaOXA-23-样或blaOXA-143基因的鲍曼不动杆菌分离株中的首次报道。分子分型揭示了分离株之间的高度遗传异质性,和多克隆传播。
    结论:遗传抗性决定因素的积累强调了严格的感染控制措施和强有力的抗菌药物管理计划以遏制多药耐药菌株的必要性。
    OBJECTIVE: To investigate the genetic profile and characterize antimicrobial resistance, including the main β-lactam antibiotic resistance genes, in Acinetobacterbaumannii isolates from a tertiary hospital in Recife-PE, Brazil, in the post-COVID-19 pandemic period.
    RESULTS: Acinetobacter baumannii isolates were collected between 2023 and 2024 from diverse clinical samples. Antimicrobial resistance testing followed standardized protocols, with β-lactamase-encoding genes detected via PCR and sequencing. Investigation into ISAba1 upstream of blaOXA-carbapenemase and blaADC genes was also conducted. Genetic diversity was assessed through ERIC-PCR. Among the 78 A. baumannii, widespread resistance to multiple antimicrobials was evident. Various acquired β-lactamase-encoding genes (blaOXA-23,-24,-58,-143, blaVIM, and blaNDM) were detected. Furthermore, this is the first report of blaVIM-2 in A. baumannii isolates harboring either the blaOXA-23-like or the blaOXA-143 gene in Brazil. Molecular typing revealed a high genetic heterogeneity among the isolates, and multi-clonal dissemination.
    CONCLUSIONS: The accumulation of genetic resistance determinants underscores the necessity for stringent infection control measures and robust antimicrobial stewardship programs to curb multidrug-resistant strains.
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  • 文章类型: Journal Article
    布鲁氏菌病,主要由melitensis布鲁氏菌(B.melitensis),在中国被认为是一种重要的人畜共患疾病。在威海,位于山东半岛的东端,布鲁氏菌病在过去五年一直处于低流行阶段。
    这是过去五年布鲁氏菌病爆发的初步报告。B.melitensisbv菌株。3来自威海与其他城市表现出亲密的亲缘关系,暗示了一个潜在的共同祖先.
    流行病学调查依赖于公共卫生实验室的标准化和有效的分子分型方法和分析工具,以识别和追踪暴发。了解受灾严重地区自由放养家庭的牲畜流通模式对于控制布鲁氏菌病的传播至关重要。
    UNASSIGNED: Brucellosis, mainly caused by Brucella melitensis (B. melitensis), is regarded as a significant zoonotic disease in China. In Weihai, located at the eastern end of the Shandong Peninsula, brucellosis has been in a low epidemic phase for the past five years.
    UNASSIGNED: This was the initial report of a brucellosis outbreak in the last five years. Strains of B. melitensis bv. 3 from Weihai and other cities showed a close genetic relationship, suggesting a potential common ancestry.
    UNASSIGNED: Epidemiological investigations depend on standardized and effective molecular typing methods and analysis tools for public health laboratories to identify and trace outbreaks. Understanding the circulation patterns of livestock in free-range households in heavily affected areas is essential for controlling the spread of brucellosis.
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  • 文章类型: English Abstract
    目的:评价脉冲场凝胶电泳(PFGE)三种分子分型方法的相关性,重复基因外回文(REP)-PCR和超细菌重复基因间共识(ERIC)-PCR,并探索菌株之间的遗传关系,结合血清型分析,进一步了解辽宁省副溶血性弧菌的分布和流行趋势。
    方法:血清分型,PFGE,REP-PCR,对2018年辽宁省150株VP分离株进行ERIC-PCR分子分型和聚类分析。
    结果:118个分离株可分为14个血清型,32个分离株无法分类。主要血清型为O3、O1和O2。PFGE的分辨率(DI)为0.969,REP-PCR的分辨率(DI)为0.948,ERIC-PCR的分辨率(DI)为0.927。血清型O3组菌株与O1组菌株的分子类型高度相似。
    结论:2018年,辽宁省临床VP分离株流行血清型仍为O3:K6,食品VP分离株流行血清型仍为O2。PFGE的结果,REP-PCR,ERIC-PCR分型方法是一致的,PFGE分型法的分辩率和重现性均优于其他两种分型法。血清型O3组与O1组密切相关。
    OBJECTIVE: To evaluate the correlation among the three molecular typing method of pulsed field gel electrophoresis(PFGE), repetitive extragenic palindromic(REP)-PCR and en-terobacterial repetitive intergenic consensus(ERIC)-PCR, and to explore the genetic relationship among strains, and to further understand the distribution and epidemic trend of Vibrio parahaemolyticus in Liaoning Province by combining Serotype analysis.
    METHODS: Serum typing, PFGE, REP-PCR, and ERIC-PCR molecular typing and cluster analysis were performed on 150 VP isolates from Liaoning Province in 2018.
    RESULTS: 118 isolates could be divided into 14 Serotype, and 32 isolates could not be classified. The main serotypes were O3, O1 and O2. The resolution(DI) of PFGE is 0.969, the resolution(DI) of REP-PCR is 0.948, and the resolution(DI) of ERIC-PCR is 0.927. The Serotype O3 group strains are highly similar to the molecular types of O1 group strains.
    CONCLUSIONS: In 2018, the epidemic Serotype of clinical VP isolates in Liaoning Province is still O3: K6, and the epidemic serotype of food VP isolates is still O2. The result of PFGE, REP-PCR, and ERIC-PCR typing method are consistent, and the resolution and reproducibility of PFGE typing method are superior to the other two method. The Serotype O3 group is closely related to O1 group.
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  • 文章类型: Multicenter Study
    加替隐球菌(C.gattii)正在全球范围内成为一种威胁生命的真菌。然而,中国的数据很少。在这项研究中,从中国多家医院收集了27个分离株和32个患者信息,加上以前报告的数据(80例),来描述C.gattii的地理分布.采用多位点序列分型对分离株进行了分子鉴定和遗传多样性分析,同时通过体外表型测试和体内动物实验探讨了微生物学和毒力特征。研究结果表明,感染了C.gattii的患者主要是具有免疫能力的男性,大部分表现出中枢神经系统受累的症状。分离菌株主要分布在热带和亚热带地区,以VGI基因型为主。这些菌株表现出明显的遗传多样性,在57个分离株中识别25个不同的序列类型(ST)(可用数据),包括三个新颖的ST(ST565、ST567和ST568)。体外测定揭示了VGI和VGII在39°C下的生长能力之间的显着差异,胶囊直径,黑色素的产生,抗紫外线,和抗氧化能力,VGII表现出更大的弹性。结合动物实验和临床预后表明,致病性与体外毒力表型或分子基因型没有直接相关,强调毒力的复杂性。此外,组织病理学分析表明,肺组织损伤可能主要导致小鼠死亡,特别是更多的致病菌株引起广泛的肺组织损伤。我们的研究有可能为全面了解中国C.gattii的微生物学特性提供有价值的数据。重要性我们的研究表明,加替隐球菌的分子分型与毒力无关。动物实验和临床预后的整合表明,致病性与体外毒力表型或分子基因型没有直接相关性。强调毒力的错综复杂的性质。总之,我们的研究有可能提供有价值的见解,以了解中国的C.gattii的微生物属性。
    OBJECTIVE: Our study indicates that the molecular typing of Cryptococcus gattii is unrelated to virulence. The integration of animal experiments and clinical prognosis demonstrated that pathogenicity did not exhibit a direct correlation with in vitro virulence phenotypes or molecular genotypes, emphasizing the intricate nature of virulence. In conclusion, our research holds the potential to provide valuable insights into understanding the microbiological attributes of C. gattii in China.
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  • 文章类型: Journal Article
    卵巢癌是一种具有不同分子表型的异质性疾病。我们使用细胞分化轨迹分析对卵巢癌进行分子分型,并提出预后风险评分模型。使用由inereCNV提供的拷贝数变异,我们确定了恶性肿瘤细胞。然后,根据分化相关基因(DRGs)将卵巢癌样本分为4种亚型.生存率有显著差异,临床特征,肿瘤微环境评分,和ICGs在亚型之间的表达水平。基于九个DRG,产生预后风险评分模型(1年AUC:0.749;3年AUC:0.651).然后我们获得了预后变量组合的列线图,包括风险评分和临床病理特征,并预测了1-,3年和5年总生存期。最后,我们利用建立的风险模型探讨了免疫逃逸的一些问题。我们的研究证明了细胞分化对预测OV患者预后的重要影响,并为OV治疗和潜在的免疫治疗策略提供了新的见解。
    Ovarian cancer is a heterogeneous disease with different molecular phenotypes. We performed molecular typing of ovarian cancer using cell differentiation trajectory analysis and proposed a prognostic risk scoring model. Using the copy number variation provided by inferCNV, we identified malignant tumor cells. Then, ovarian cancer samples were divided into four subtypes based on differentiation-related genes (DRGs). There were significant differences in survival rates, clinical features, tumor microenvironment scores, and the expression levels of ICGs among the subtypes. Based on nine DRGs, a prognostic risk score model was generated (AUC at 1 year: 0.749; 3 years: 0.651). Then we obtained a nomogram of the prognostic variable combination, including risk scores and clinicopathological characteristics, and predicted the 1-, 3- and 5-year overall survival. Finally, we explored some issues of immune escape using the established risk model. Our study demonstrates the significant influence of cell differentiation on predicting prognosis in OV patients and provides new insights for OV treatment and potential immunotherapeutic strategies.
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  • 文章类型: Journal Article
    弥漫性大B细胞淋巴瘤(DLBCL)是一种高度异质性的非霍奇金淋巴瘤,在临床和遗传方面。诚然,DLBCL按遗传学分为六种亚型,含有MCD,BN2,EZB,N1、ST2和A53。血脂异常与多种实体瘤有关,最近有报道称与血液恶性肿瘤有关。我们的目的是提出一项基于分子亚型的DLBCL血脂异常的回顾性研究。
    这项研究得出结论,259例新诊断的DLBCL患者及其活检标本可用于分子分型。结果表明,血脂异常的发生率(87.0%,p<0.001)在EZB亚型中高于其他亚型,尤其是高甘油三酯血症(78.3%,在EZB亚型中p=0.001)。基于病理基因测序,BCL2基因融合突变患者与高脂血症显著相关(76.5%,p=0.006)和高甘油三酯血症(88.2%,p=0.002)。然而,血脂异常的发生对预后无显著影响。
    总之,血脂异常与DLBCL的遗传异质性相关,对生存无显著影响.这项研究首先将DLBCL中的脂质和遗传亚型联系起来。
    UNASSIGNED: Diffuse large B-cell lymphoma (DLBCL) is a kind of highly heterogeneous non-Hodgkin lymphoma, both in clinical and genetic terms. DLBCL is admittedly categorized into six subtypes by genetics, which contain MCD, BN2, EZB, N1, ST2, and A53. Dyslipidemia is relevant to a multitude of solid tumors and has recently been reported to be associated with hematologic malignancies. We aim to present a retrospective study investigating dyslipidemia in DLBCL based on the molecular subtypes.
    UNASSIGNED: This study concluded that 259 patients with newly diagnosed DLBCL and their biopsy specimens were available for molecular typing. Results show that the incidence of dyslipidemia (87.0%, p <0.001) is higher in the EZB subtype than in others, especially hypertriglyceridemia (78.3%, p = 0.001) in the EZB subtype. Based on the pathological gene-sequencing, patients with BCL2 gene fusion mutation are significantly correlative with hyperlipidemia (76.5%, p = 0.006) and hypertriglyceridemia (88.2%, p = 0.002). Nevertheless, the occurrence of dyslipidemia has no remarkable influence on prognosis.
    UNASSIGNED: In summary, dyslipidemia correlates with genetic heterogeneity in DLBCL without having a significant influence on survival. This research first connects lipids and genetic subtypes in DLBCL.
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  • 文章类型: Journal Article
    目标:手,口蹄疫(HFMD)是一种传染性的儿科病毒性疾病,是由于小牛科的肠道病毒(EV)引起的。一家三级保健中心报告了手足口病病例,乌德汉普尔,(查谟和克什米尔),印度北部。本研究强调了HFMD病例的临床和分子病毒学方面。
    方法:纳入2016年8月至2017年9月期间报告的、临床诊断为HFMD的所有年龄组的病例。临床,比较了生化和分子病毒学方面。临床样本(n=50),如囊泡拭子,收集口腔和咽拭子进行肠道病毒检测。通过基于5'NCR的RT-PCR检测EV-RNA,并通过VP1基因扩增和循环测序进行基因分型。
    结果:在登记的手足口病病例中(n=50),最高(84%)的是5岁以下的儿童,表现出一种或两种症状都有前驱症状(发烧,烦躁)。临床表现主要涉及嘴唇和舌头上的口腔溃疡(48%)。口腔糜烂为单个或多个。在手上和手掌上都能看到exanthemas,广泛传播到臀部,腿,手臂和躯干。其中,六名患者被发现贫血。全血细胞计数(CBC)显示5岁以下儿童的淋巴细胞增多和C反应蛋白(n=10)。在78%(39/50)的临床样品中检测到EV-RNA。基于VP1基因的分型表明存在CV-A16,CVA6和EV-A71类型。
    结论:该研究强调了报告地区手足口病病例中EVs的相关性。Udhampur(J&K)首次报道了CV-A16,CV-A6和EV-A71类型,印度北部。在检测到的EV菌株的临床特征中未观察到差异。菌株的传播保证和警报爆发。对手足口病和病毒株的监测更有重点的研究是强制性的。
    Hand, Foot and Mouth disease (HFMD) is a contagious pediatric viral disease caused due to enteroviruses (EV) of the family Picornaviridae. Cases of HFMD were reported from a tertiary care health centre, Udhampur, (Jammu and Kashmir), Northern India. The present study highlights the clinical and molecular virological aspects of HFMD cases.
    Cases reported during August 2016-September 2017, and clinically diagnosed as HFMD of all age groups were included. Clinical, Biochemical and molecular virology aspects were compared. Clinical samples (n ​= ​50) such as vesicle swab, buccal and throat swabs were collected for enterovirus detection. EV-RNA was detected by 5\'NCR based RT-PCR and genotyping by VP1 gene amplification and cycle sequencing.
    Of the cases of HFMD enrolled (n ​= ​50), highest (84%) were of children aged <5 years, presented either or both anathemas and exanthemas with prodromal symptoms (fever, irritability). Clinical presentations involved mainly oral ulcers on lips and tongue (48%). Oral erosions were either single or multiple in numbers. Exanthemas were seen on hand and palm, widely spread up to buttocks, legs, arms and trunk. Of these, six patients were found anemic. Complete blood count (CBC) indicated lymphocytosis and C-reactive protein (n ​= ​10) in children aged <5 years. EV-RNA was detected in 78% (39/50) of the clinical samples. VP1 gene based typing indicated the presence of CV-A16, CVA6 and EV-A71 types.
    The study highlights association of EVs in HFMD cases in the reported region. CV-A16, CV-A6 and EV-A71 types were reported for the first time from Udhampur (J&K), Northern India. No differences were observed in the clinical profile of EV strains detected. Circulation of the strains warrant and alarm outbreaks. More focused studies on HFMD and monitoring of viral strains is mandatory.
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  • 文章类型: Journal Article
    目的:研究脑胶质瘤O6-甲基鸟嘌呤DNA甲基转移酶(MGMT)启动子甲基化状态对O-(2-18F-氟乙基)-L-酪氨酸([18F]FET)摄取和动脉自旋标记(ASL)脑血流量(CBF)的影响。通过混合PET/MR评估。立体定向活检用于验证发现。
    方法:在57例新诊断的神经胶质瘤患者中,根据PET/FLAIR成像描绘了一组完整的肿瘤和感兴趣的参考体积(VOIs),并转移到相应的[18F]FETPET和CBF图。计算平均和最大肿瘤-脑比率(TBR)和标准化CBF(nCBF)。[18F]FETPET和CBF在确定胶质瘤MGMT启动子甲基化状态中的预测功效通过全肿瘤分析和立体定向活检来评估。使用从多个立体定向活检获得的组织学标本分析PET/MR参数与MGMT启动子甲基化之间的相关性。
    结果:基于对整个肿瘤体积和活检部位的分析,TBRmean,TBRmax,nCBFmean,和nCBFmax在有和没有MGMT启动子甲基化的胶质瘤之间没有统计学意义(均p>0.05)。此外,立体定向活检显示TBRmean,TBRmax,nCBFmean,nCBFmax与MGMT启动子甲基化无相关性(r=-0.117,p=0.579;r=-0.161,p=0.443;r=-0.271,p=0.191;r=-0.300,p=0.145)。
    结论:MGMT启动子甲基化状态对胶质瘤中ASL的[18F]FET摄取和CBF无影响。立体定向活检证实了这一点,并进一步揭示了[18F]FETPET摄取和CBF与MGMT启动子甲基化的百分比没有相关性。
    结论:•基于整个肿瘤VOI评估,MGMT启动子甲基化状态对胶质瘤中ASL的[18F]FET摄取和CBF没有影响。•对于WHOIV级胶质母细胞瘤,基于混合PET/MR的[18F]FETPET和ASL参数无法预测MGMT启动子甲基化状态。•基于立体定向图像的组织学显示[18F]FETPET摄取和CBF与胶质瘤中MGMT启动子甲基化的状态和百分比没有相关性。
    OBJECTIVE: To investigate the effects of O6-methylguanine DNA methyltransferase (MGMT) promoter methylation status of gliomas on O-(2-18F-fluoroethyl)-L-tyrosine ([18F]FET) uptake and cerebral blood flow (CBF) of arterial spin labeling (ASL), evaluated by hybrid PET/MR. Stereotactic biopsy was used to validate the findings.
    METHODS: A set of whole tumor and reference volumes of interest (VOIs) based on PET/FLAIR imaging were delineated and transferred to the corresponding [18F]FET PET and CBF maps in 57 patients with newly diagnosed gliomas. The mean and max tumor-to-brain ratio (TBR) and normalized CBF (nCBF) were calculated. The predictive efficacy of [18F]FET PET and CBF in determining MGMT promoter methylation status of glioma were evaluated by whole tumor analysis and stereotactic biopsy. The correlation between PET/MR parameters and MGMT promoter methylation were analyzed using histological specimens acquired from multiple stereotactic biopsies.
    RESULTS: Based on the analysis of whole tumor volume and biopsy site, TBRmean, TBRmax, nCBFmean, and nCBFmax showed no statistically significant differences between gliomas with and without MGMT promoter methylation (all p > 0.05). Furthermore, stereotactic biopsy demonstrated that TBRmean, TBRmax, nCBFmean, and nCBFmax showed no correlation with MGMT promoter methylation (r = -0.117, p = 0.579; r = -0.161, p = 0.443; r = -0.271, p = 0.191; r = -0.300, p = 0.145; respectively).
    CONCLUSIONS: MGMT promoter methylation status shows no effect on [18F]FET uptake and CBF of ASL in gliomas. Stereotactic biopsy validates it and further reveals there is no correlation of [18F]FET PET uptake and CBF with the percentages of MGMT promoter methylation.
    CONCLUSIONS: • Based on whole tumor VOI assessment, MGMT promoter methylation status shows no effect on [18F]FET uptake and CBF of ASL in gliomas. • For WHO grade IV glioblastomas, [18F]FET PET and ASL parameters based on hybrid PET/MR fail to predict the MGMT promoter methylation status. • Stereotactic image-based histology reveals that there is no correlation of [18F]FET PET uptake and CBF with the status and percentages of MGMT promoter methylation in gliomas.
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  • 文章类型: Journal Article
    Aspergillus exhibits a wide variation of susceptibility against antifungals according to genetic and environmental factors. Identification to the species level is necessary for appropriate treatment. Our objective was to determine the Aspergillus species involved in invasive pulmonary aspergillosis (IPA) among ICU patients in Jakarta, Indonesia.
    The incidence of IPA in ICU patients at six hospitals in Jakarta from October 2012 - January 2015 was investigated. It involved a collection of endotracheal aspirates (ETA), nasal swabs and environmental samples around the hospitals, phenotypic screening, molecular characterization, and antifungal susceptibility testing.
    Of the 405 patients investigated, 31 patients (7.7%) were diagnosed with putative IPA, from whom 45 Aspergillus isolates were collected. Aspergillus isolates were identified from pulmonary secretions in 24 patients, from nasal swabs in 7 patients and from both pulmonary secretions and nasal swabs in 7 patients. The phenotypic method showed 33 isolates of Aspergillus flavus (73.4%), nine Aspergillus fumigatus (20%), two Aspergillus niger (4.4%), and one Aspergillus nidulans (2.2%) isolate. Molecular identification showed 27 isolates of A. flavus (60.0%), eight isolates of A. fumigatus (17.8%), two isolates of A. niger (4.4%) and one isolate of A. nidulans (2.2%), while seven isolates (15.6%) were cryptic species or mixed isolates.
    Susceptibility testing showed all isolates were susceptible to amphotericin B, azoles and micafungin. Aspergillus flavus was the main causative organism in IPA cases in Jakarta, followed by A. fumigatus.
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  • 文章类型: Journal Article
    布鲁氏菌病是许多发展中国家的地方病,被世界卫生组织列为“被忽视的人畜共患病”前七名。尽管1998年启动了巴勒斯坦布鲁氏菌病控制计划,但该疾病在2012年后再次出现。有趣的是,据报道,邻近的以色列地区也出现了类似的重新出现模式。这项工作的目的是表征重新出现的菌株并描述它们的遗传相关性。在2015年至2017年期间,使用两项血清学测试对1324例疑似人类病例的血液样本进行了分析。血清阳性样本进行培养,通过不同的遗传标记分析它们的DNA,以确定涉及的布鲁氏菌物种,并排除任何可能涉及的Rev.1疫苗株。对9个分离株的rpoB基因进行测序,以筛选利福平抗性突变。使用多位点VNTR分析(MLVA-16)对分离物进行基因分型。分子分析表明,所有分离株均为与Rev.1疫苗无关的布鲁氏菌菌株。rpoB基因序列显示四个与利福平抗性无关的单核苷酸变异(SNV)。MLVA-16分析将分离株聚集为属于东地中海谱系的22种独特基因型。总之,我们的研究结果表明,布鲁氏菌病的重新出现是由于当地的B.melitensis菌株,巴勒斯坦和以色列控制计划的弱点可能是疾病重新出现的主要因素。然而,必须调查其他社会经济和环境因素。此外,加强布鲁氏菌病控制计划和加强所有利益相关者之间的合作对于确保抗击布鲁氏菌病的长期计划成果至关重要。
    Brucellosis is an endemic disease in many developing countries and ranked by the World Health Organization among the top seven \"neglected zoonoses\". Although a Palestinian brucellosis control program was launched in 1998, the disease re-emerged after 2012. Interestingly, a similar re-emerging pattern was reported in the neighbouring Israeli regions. The aim of this work was to characterize the re-emerging strains and delineate their genetic relatedness. During 2015-2017, blood samples from 1324 suspected human cases were analyzed using two serological tests. Seropositive samples were cultured, and their DNAs were analyzed by different genetic markers to determine the involved Brucella species and rule out any possible involvement of the Rev.1 vaccine strain. The rpoB gene was sequenced from nine isolates to screen for rifampicin resistance mutations. Multi locus VNTR analysis (MLVA-16) was used for genotyping the isolates. The molecular analysis showed that all isolates were Brucella melitensis strains unrelated to the Rev.1 vaccine. The rpoB gene sequences showed four single nucleotide variations (SNVs) not associated with rifampicin resistance. MLVA-16 analysis clustered the isolates into 22 unique genotypes that belonged to the East Mediterranean lineage. Altogether, our findings show that the re-emergence of brucellosis was due to B. melitensis strains of local origin, the Palestinian and Israeli control programs\' weaknesses could be a major factor behind the re-emergence of the disease. However, other socioeconomic and environmental factors must be investigated. Moreover, strengthening brucellosis control programs and enhancing cooperation between all stakeholders is essential to ensure long-term program outcomes to fight brucellosis.
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