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Monoclonal antibody techniques. 50 years of development 单克隆抗体技术。50 年的发展历程
Pub Date : 2024-07-16 DOI: 10.36233/0372-9311-486
Yulia S. Massino, Y. Tarakanova, Olga L. Segal, A. Pechelyulko, D. A. Yakovleva, Maria V. Lichutina, D. Dmitriev, Alexander D Dmitriev
Monoclonal antibodies are widely used in all fields of biology and medicine. The emergence and development of the technology for their production revolutionized immunology and allowed the creation of not only new diagnostic methods, but also many effective drugs. The purpose of our review is to analyze and summarize relevant data concerning the technology of obtaining monoclonal antibodies. We have analyzed information from 70 modern literary sources devoted to various methods of obtaining them. In this review, we tried to cover the entire range of methods used to obtain monoclonal antibodies today.
单克隆抗体广泛应用于生物学和医学的各个领域。单克隆抗体生产技术的出现和发展彻底改变了免疫学,不仅创造了新的诊断方法,还创造了许多有效的药物。我们的综述旨在分析和总结有关单克隆抗体获取技术的相关数据。我们分析了 70 种现代文献资料中有关获取单克隆抗体的各种方法的信息。在这篇综述中,我们试图涵盖当今用于获取单克隆抗体的所有方法。
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引用次数: 0
Changes in the etiological structure of severe acute respiratory viral infections in children and adults under the influence of the COVID-19 pandemic COVID-19 大流行影响下儿童和成人严重急性呼吸道病毒感染病原学结构的变化
Pub Date : 2024-07-16 DOI: 10.36233/0372-9311-532
A. Sominina, Darya M. Danilenko, A. Komissarov, M. M. Pisareva, T. Musaeva, K. Stolyarov, O. I. Afanasyeva, Veronika S. Timonina, Evgeny V. Venev, T. Levanyuk, E. Smorodintseva, O. Kurskaya, Alexander M. Shestopalov, E. V. Lelenkova, A. V. Semenov, D. A. Lioznov
Introduction. The traditional surveillance system for influenza and ARVI provides a general description of epidemics, but does not provide information on the age-related characteristics of the etiology and clinical peculiarities of severe acute respiratory diseases (SARI) in hospitalized patients. Aim. To monitor the etiology of SARI in hospitalized children and adults, assessing the impact of the COVID-19 pandemic on this process. Materials and methods. Standardized clinical and laboratory monitoring of SARI among 18,458 hospitalized patients was carried out in hospitals in three cities of Russia with weekly PCR detection of 11 types of pathogens. Results. According to the investigation of hospitalized patients with SARI for the period from 2018 to 2023, the viral etiology of respiratory diseases was deciphered in 58.3% of cases. Weekly monitoring showed a change in the etiological mosaic of SARI pathogens during the SARS-CoV-2 pandemic with a sharp decrease in the frequency of detection of influenza and respiratory syncytial virus (RSV) during the 2020–2021 season against the background of a significant increase of metapneumovirus and rhinovirus infections in children. During the 2022-2023 season an increase in the proportion of RSV infection in children under 6 years of age (up to 36.2%) was noted against the background of a significant decrease in the frequency of SARS-CoV-2. In the intensive care units (ICU), RSV infection was most often in children during the post-pandemic period (up to 30.1–53.6% of positive cases, p 0.001); in adults, SARS-CoV-2 was mostly detected (76,5–100% of cases, p 0.001). Conclusion. Hospital surveillance data significantly complements the epidemiological information obtained in the traditional surveillance system. Monitoring of infections has shown a continuously changing etiological infrastructure of SARI, with the disappearance of influenza and RSV during the COVID-19 pandemic and their return to circulation in the post-pandemic period.
导言。传统的流感和急性呼吸道感染监测系统能提供流行病的一般描述,但无法提供住院病人严重急性呼吸道疾病(SARI)的病因学年龄相关特征和临床特殊性的信息。目的是监测住院儿童和成人 SARI 的病因,评估 COVID-19 大流行对这一过程的影响。材料和方法。在俄罗斯三个城市的医院对 18,458 名住院患者的 SARI 进行了标准化临床和实验室监测,每周对 11 种病原体进行 PCR 检测。结果。根据对2018年至2023年期间SARI住院患者的调查,58.3%的病例破译了呼吸道疾病的病毒病因。每周监测结果显示,SARS-CoV-2 大流行期间,SARI 病原体的病原镶嵌发生了变化,在儿童偏肺病毒和鼻病毒感染显著增加的背景下,流感和呼吸道合胞病毒(RSV)的检测频率在 2020-2021 年期间急剧下降。在 2022-2023 年期间,6 岁以下儿童感染 RSV 的比例有所上升(高达 36.2%),而 SARS-CoV-2 的感染率则显著下降。在大流行后期间,重症监护室(ICU)中儿童感染 RSV 的比例最高(阳性病例比例高达 30.1-53.6%,P 0.001);而在成人中,SARS-CoV-2 的检出率最高(76.5-100%,P 0.001)。结论医院监测数据极大地补充了传统监测系统所获得的流行病学信息。对感染的监测表明,SARI 的病原体基础结构在不断变化,在 COVID-19 大流行期间,流感和 RSV 已经消失,而在大流行后,它们又重新流行起来。
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引用次数: 0
Development of a technique for molecular typing of Bacillus anthracis strains using new VNTR and INDEL markers 利用新的 VNTR 和 INDEL 标记开发炭疽杆菌菌株分子分型技术
Pub Date : 2024-07-16 DOI: 10.36233/0372-9311-487
G. Pechkovskii, E. I. Eremenko, A. Ryazanova, S. Pisarenko, N. Shapakov, L. Aksenova, O. V. Semenova, Lyudmila D. Timchenko, A. Kulichenko
Introduction. Bacillus anthracis, the pathogen of a particularly dangerous zoonotic disease known as anthrax, requires strict epidemiological control and is characterized by high genetic homogeneity, which necessitates the development of genotyping methods. The aim of the study were to to find and characterize the VNTR and INDEL loci of B. anthracis and to develop on their basis a genotyping technique by PCR with electrophoretic detection of the results. Materials and methods. Marker search and phylogenetic analysis were performed on a sample of 388 genomes of B. anthracis strains, 322 from the GenBank collection (RefSeq) and 66 from the collection of the Stavropol Anti-Plague Institute of Rospotrebnadzor. Phylogenetic analysis was performed on the basis of SNP crustal alignment using the Parsnp program. The search for markers was carried out using the Mauve program and author's scripts in Python. PCR was performed using a ScreenMix-HS kit (CJSC "Eurogen", Russia). Results. Genomic variations of B. anthracis strains (SNP — 25,664, SNR — 14,387, VNTR — 693, INDEL — 14,667) were found, bioinformatic analysis of which revealed nine new VNTR and six INDEL molecular markers most suitable for genotyping. The genetic (allelic) variants of the markers are described. Primers were selected for the found markers and a PCR protocol with detection by electrophoresis in agarose gel was developed. When typing using VNTR markers was applied, the strains were divided into nine clusters: A.Br.Ames, A.Br.001/002, A.Br.Aust94, A.Br.005/006, A.Br.008/009 (Tsiankovskii), A.Br.008/009 (STI), A.Br.008/009 (A.Br.125), A.Br.008/009 (strain 228/269), B.Br.001/002. When typing using INDEL markers, the strains were divided into six clusters: A.Br.Ames, A.Br.001/002, A.Br.Aust94, A.Br.008/009(Tsiankovskii), B.Br.001/002(B.Br.014), as well as a cluster comprising several genetic lineages: A.Br.008/009 (STI), A.Br.008/009 (A.Br.125), A.Br.005/006 и B.Br.001/002. Conclusion. The use of the developed methodology for the identification of variable VNTR and INDEL loci makes it possible to reliably determine the phylogenetic position of B. anthracis strains and is promising for use in the epidemiological investigation of anthrax outbreaks.
导言。炭疽杆菌(Bacillus anthracis)是一种特别危险的人畜共患疾病--炭疽的病原体,需要严格的流行病学控制,其特点是具有高度的遗传同质性,因此有必要开发基因分型方法。这项研究的目的是找到炭疽杆菌的 VNTR 和 INDEL 基因位点并确定其特征,在此基础上开发一种通过 PCR 进行基因分型的技术,并对结果进行电泳检测。材料和方法对 388 个炭疽杆菌菌株基因组样本进行了标记搜索和系统进化分析,其中 322 个来自 GenBank 库(RefSeq),66 个来自 Rospotrebnadzor 的斯塔夫罗波尔抗鼠疫研究所。系统发育分析是在使用 Parsnp 程序进行 SNP 地壳比对的基础上进行的。标记搜索使用 Mauve 程序和作者的 Python 脚本进行。PCR 使用 ScreenMix-HS 试剂盒(CJSC "Eurogen",俄罗斯)进行。结果发现了炭疽杆菌菌株的基因组变异(SNP - 25,664 个,SNR - 14,387 个,VNTR - 693 个,INDEL - 14,667 个),通过生物信息学分析发现了最适合进行基因分型的 9 个新的 VNTR 和 6 个 INDEL 分子标记。文中描述了这些标记的基因(等位基因)变异。为所发现的标记选择了引物,并制定了通过琼脂糖凝胶电泳进行检测的 PCR 方案。在使用 VNTR 标记进行分型时,菌株被分为九个群:A.Br.Ames、A.Br.001/002、A.Br.Aust94、A.Br.005/006、A.Br.008/009(Tsiankovskii)、A.Br.008/009(STI)、A.Br.008/009(A.Br.125)、A.Br.008/009(228/269 株)、B.Br.001/002。在使用 INDEL 标记进行分型时,菌株被分为六个群组:A.Br.Ames、A.Br.001/002、A.Br.Aust94、A.Br.008/009(Tsiankovskii)、B.Br.001/002(B.Br.014),以及一个由多个遗传系组成的群:A.Br.008/009(STI)、A.Br.008/009(A.Br.125)、A.Br.005/006 и B.Br.001/002。结论使用所开发的方法鉴定可变 VNTR 和 INDEL 位点,可以可靠地确定炭疽杆菌菌株的系统发育位置,有望用于炭疽疫情的流行病学调查。
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引用次数: 0
Detection and type identification of non-polio enteroviruses in children against the background of acute intestinal infections of various etiologies: 2018–2023 以各种病因引起的急性肠道感染为背景的儿童非脊髓灰质炎肠道病毒的检测和类型鉴定:2018-2023 年
Pub Date : 2024-07-16 DOI: 10.36233/0372-9311-513
S. Selivanova, Vladimir V. Zverev, N. Ponomareva, AV Leonov, A. Y. Kashnikov, N. V. Epifanova, A. V. Polyanina, N. Novikova
Introduction. Enteroviruses (EV) are characterized by: species and type diversity, polymorphism of clinical manifestations, a tendency to epidemic spread, and are often the cause of disease outbreaks, which determines the relevance of monitoring EV strains in various clinical forms of infection, including in conditions of anti-epidemic measures. The aim of the study: to characterize the prevalence and diversity of non-polio enteroviruses (NPEV) types in children with acute intestinal infection (AII) in the period 2018–2023, including the COVID-19 pandemic. Materials and methods. The RT-PCR method was used to study 7302 samples of feces from children hospitalized with a diagnosis of AII in the infectious diseases hospital of Nizhny Novgorod. Genotyping of EV strains was carried out using fragment Sanger sequencing of the genome region encoding capsid protein 1 (VP1) and the online resource BLAST. Results. EVs were found in 5.0 ± 0.3% (1.7–7.8%), both in mono- and mixed infections with other enteric viruses. The long-term dynamics of the frequency of EV detection and the incidence of EV infection in children in the Nizhny Novgorod region was characterized by a sharp decrease in indicators in 2020 against the backdrop of the introduction of anti-epidemic measures. When genotyping 299 strains, 41 types of NPEV of 4 species were identified. The spectrum included the main pathogens of exanthema and neuroinfections and rare types found in “minor” or intestinal forms of infection. During the study period, a redistribution of NPEV species was established. Before the pandemic, the ratio of Enterovirus A : Enterovirus B : Enterovirus C species was as follows — 41.0 : 46.7 : 12.3%; during the 2020 pandemic season the ratio was 0.0 : 37.5 : 62.5%; after the lifting of restrictive measures — 47 : 29 : 23%, which may be due to the different effectiveness of the restrictive measures on the mechanisms of transmission of EVs of different types. Conclusion. The genetic diversity of NPEVs detected in children with AII complements information on the typical composition of the territorial enterovirus population. In children with AII, when the airborne transmission of SARS-CoV-2 was blocked, there was a decrease in the frequency of detection of viruses of the Enterovirus B type, the absence of detection of Enterovirus A and the constant presence of Enterovirus C.
导言。肠道病毒(EV)的特点是:种类和类型的多样性、临床表现的多态性、流行传播的倾向性,并且往往是疾病暴发的原因,这就决定了在各种临床感染形式中监测 EV 株系的相关性,包括在采取抗流行措施的条件下。本研究的目的:描述 2018-2023 年期间急性肠道感染(AII)患儿中非脊髓灰质炎肠道病毒(NPEV)类型的流行率和多样性,包括 COVID-19 大流行。材料与方法。采用 RT-PCR 方法研究了 7302 份下诺夫哥罗德市传染病医院诊断为 AII 的住院儿童粪便样本。通过对编码帽状蛋白 1(VP1)的基因组区域进行片段 Sanger 测序,并利用在线资源 BLAST 对 EV 株进行基因分型。结果发现在单一病毒感染和与其他肠道病毒混合感染中,发现了 5.0 ± 0.3% (1.7-7.8%)的 EV。下诺夫哥罗德地区儿童中发现 EV 的频率和 EV 感染率的长期动态特点是,在采取抗疫情措施的背景下,各项指标在 2020 年急剧下降。在对 299 株菌株进行基因分型时,发现了 4 个物种的 41 种 NPEV。其谱系包括外感和神经感染的主要病原体,以及在 "小 "感染或肠道感染中发现的罕见类型。在研究期间,确定了 NPEV 物种的重新分布。在大流行之前,肠道病毒 A:肠道病毒 B:肠道病毒 C 的比例为 41.0:46.7:12.3%;在 2020 年大流行季节,该比例为 0.0:37.5:62.5%;在取消限制措施之后,该比例为 47:29:23%,这可能是由于限制措施对不同类型 EV 传播机制的效果不同。结论在 AII 患儿中检测到的 NPEVs 基因多样性补充了有关地区肠道病毒群典型组成的信息。在患有 AII 的儿童中,当 SARS-CoV-2 的空气传播被阻断时,检测到 B 型肠道病毒的频率降低,没有检测到 A 型肠道病毒,而 C 型肠道病毒持续存在。
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引用次数: 0
Hospital environment microbiome 医院环境微生物群
Pub Date : 2024-07-16 DOI: 10.36233/0372-9311-520
E. B. Brusina
The aim of the review is to give a brief description of the biodiversity and structure of the hospital environment microbiome based on molecular genetic research methods. Until a certain time, studies of the hospital environment microbiota for the purposes of epidemiological surveillance and control of healthcare-associated infections (HAIs) were based on routine microbiological identification of clinically relevant bacterial taxa. Discovery of DNA, the development of sequencing technologies, PCR and cloning techniques enabled the investigation of microbial communities using cultivation-independent, DNA and RNA-based approaches. At the current level of knowledge, the hospital environment can be considered as a superorganism with its own microbiome. Multiomic technologies, including meta-transcriptomic, meta-proteomic and metabolomic approaches, provide detailed information about microbial activity in the environment. Now it has been established that there is a stable core of the hospital microbiome where the vast majority of microorganisms are necessary for the functioning of the hospital ecosystem and are not classified as human pathogens. The hospital microbiome has a homogeneous structure composed by a massive dominance of a few taxa and microbial network with low connectivity forming a clustered topology. A keystone species is a taxon whose importance for maintaining community structure is relatively higher than others and its identification is of paramount importance. Due to the lack of knowledge of the hospital environment microbiome by molecular genetic technologies, there is no single shared point of view on the microbial diversity in different healthcare facilities. But there is no doubt that molecular genetic technologies will shed light on the evolution of hospital strains and determine which indicators are the most informative for monitoring and prognosis of HAIs.
本综述旨在根据分子遗传学研究方法简要介绍医院环境微生物群的生物多样性和结构。在此之前,以流行病学监测和控制医疗相关感染(HAIs)为目的的医院环境微生物群研究都是基于对临床相关细菌类群的常规微生物鉴定。随着 DNA 的发现、测序技术、PCR 和克隆技术的发展,人们可以利用不依赖培养、基于 DNA 和 RNA 的方法对微生物群落进行调查。就目前的知识水平而言,医院环境可被视为一个拥有自己微生物群的超级有机体。多组学技术,包括元转录组学、元蛋白组学和代谢组学方法,提供了环境中微生物活动的详细信息。现在已经确定,医院微生物组有一个稳定的核心,其中绝大多数微生物是医院生态系统运作所必需的,不属于人类病原体。医院微生物群具有同质结构,由少数几个类群和连接性较低的微生物网络组成,形成一个聚类拓扑结构。关键物种是指对维持群落结构的重要性相对高于其他物种的类群,因此对其进行鉴定至关重要。由于缺乏通过分子基因技术对医院环境微生物组的了解,目前还没有关于不同医疗机构微生物多样性的统一观点。但毫无疑问,分子基因技术将揭示医院菌株的演变过程,并确定哪些指标对 HAIs 的监测和预后最有参考价值。
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引用次数: 0
Molecular characteristics of fluoroquinolone-resistant Mycobacterium tuberculosis strains from newly diagnosed tuberculosis patients in the Northwest of Russia 俄罗斯西北部新诊断出的结核病人中耐氟喹诺酮结核分枝杆菌菌株的分子特征
Pub Date : 2024-07-16 DOI: 10.36233/0372-9311-510
A. Vyazovaya, N. S. Solovieva, A. Gerasimova, V. Zhuravlev, I. Mokrousov
Introduction. Fluoroquinolones remain the key second-line anti-tuberculosis drugs. The aim of the study was the molecular characterization of fluoroquinolone-resistant Mycobacterium tuberculosis strains from newly diagnosed tuberculosis patients in the Northwest of the Russian Federation. Materials and methods. The retrospective study collection included M. tuberculosis isolates isolated in 2015–2019 from previously untreated tuberculosis patients. Susceptibility to antituberculosis drugs (including the fluoroquinolone ofloxacin) was determined using the BACTEC MGIT960 or absolute concentration method. Mutations in the gyrA gene as a marker of resistance to fluoroquinolones, were detected by real-time PCR. Beijing genotype and its subtypes were detected by PCR and real-time PCR methods. Non-Beijing strains were spoligotyped. Results and discussion. Phenotypic resistance to ofloxacin was detected in 6.7% (40/599) of strains and in 17.4% (40/230) of MDR strains. 34 of 40 (85%) ofloxacin-resistant strains belonged to the Beijing genotype. 18 (45%) strains were assigned to the Russian epidemic subtype Beijing B0/W148 and 12 (30%) to Beijing Central Asian/Russian. The remaining 6 ofloxacin-resistant strains belonged to the Euro-American phylogenetic lineage. Mutations in the gyrA gene were found in 97.5% (39/40) of strains. The most common were mutations in codon 94 (69.2%, 27/39). The Asp94Gly substitution was identified in 57.5% (23/40) of ofloxacin-resistant strains and was dominant among Beijing (19/34) and non-Beijing (4/6) strains. The second most common substitution was Ala90Val (25%, 10/40). More than half of the ofloxacin-resistant strains, Beijing B0/W148 (10/18) and Central Asian/Russian (7/12), carried the Asp94Gly mutation. Conclusion. In the Northwest of Russia in 2016-2019, primary resistance of M. tuberculosis to fluoroquinolones was 6.7% in the total collection and 17.4% of MDR strains, and was mainly caused by the gyrA Asp94Gly and Ala90Val mutations. Beijing B0/W148 genotype was characterized by the largest proportion of fluoroquinolone-resistant strains.
导言。氟喹诺酮类药物仍然是主要的二线抗结核药物。本研究的目的是对俄罗斯联邦西北部新诊断的结核病患者中耐受氟喹诺酮类药物的结核分枝杆菌菌株进行分子鉴定。材料和方法。回顾性研究收集的样本包括 2015-2019 年从既往未经治疗的肺结核患者中分离出的结核分枝杆菌。对抗结核药物(包括氟喹诺酮类药物氧氟沙星)的敏感性采用 BACTEC MGIT960 或绝对浓度法进行测定。gyrA基因突变是对氟喹诺酮类药物产生耐药性的标志,通过实时聚合酶链式反应(real-time PCR)进行检测。通过 PCR 和实时 PCR 方法检测北京基因型及其亚型。对非北京菌株进行了分离分型。结果与讨论6.7%的菌株(40/599)和17.4%的MDR菌株(40/230)对氧氟沙星具有表型耐药性。40 株对氧氟沙星耐药的菌株中有 34 株(85%)属于北京基因型。18株(45%)属于俄罗斯流行亚型北京 B0/W148,12株(30%)属于北京中亚/俄罗斯。其余 6 株对氧氟沙星耐药的菌株属于欧美系统发育系。在 97.5%(39/40)的菌株中发现了 gyrA 基因突变。最常见的是密码子94的突变(69.2%,27/39)。在 57.5%(23/40)的耐氧菌株中发现了 Asp94Gly 突变,在北京菌株(19/34)和非北京菌株(4/6)中占优势。其次是 Ala90Val(25%,10/40)。一半以上的耐氧氟沙星菌株(北京 B0/W148(10/18)和中亚/俄罗斯(7/12))携带 Asp94Gly 突变。结论2016-2019年,俄罗斯西北部地区结核杆菌对氟喹诺酮类药物的原发性耐药菌株占菌株总数的6.7%,MDR菌株占17.4%,主要由gyrA Asp94Gly和Ala90Val突变引起。北京 B0/W148 基因型的耐氟喹诺酮菌株比例最高。
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引用次数: 0
Campylobacteriosis: genotypic characteristics of the pathogen and immunological status of patients 弯曲杆菌病:病原体的基因型特征和患者的免疫状况
Pub Date : 2024-07-16 DOI: 10.36233/0372-9311-531
Yury V. Lobzin, K. Ermolenko, Mariia A. Makarova, Lidiya A. Kaftyreva, Elvira A. Martens, Dmitriy E. Polev, E.I. Ermolenko
Introduction. Campylobacteriosis is among the leading causes of acute gastrointestinal infections. The severity of campylobacteriosis and the development of long-term complications may be influenced by the genotype of the pathogen, whose biological properties can affect immune response parameters. The aim of the study was to identify common genotypes of epidemic clones of Campylobacter pathogens and to investigate characteristics of the immune response and severity of the disease. Materials and methods. The study included 203 patients aged from 1 month to 17 years with campylobacteriosis who underwent treatment at the clinic of the Federal State Budgetary Institution "DNKCIB FMBA" in 2019–2021. The diagnosis was confirmed using polymerase chain reaction method. Patient samples were also analyzed using culture-based methods. Total DNA was extracted using the QIAamp DNA Mini Kit. Genetic determinants encoding virulence factors and MLST typing were performed using the ResFinder program. The immune status of patients was assessed on days 1 and 7 of the illness. Immunological investigation included measurement of serum immunoglobulin concentrations (IgA, IgM, IgG), C-reactive protein, and cytokines (IL-1β, IL-1, IL-2, IL-4, IL-5, IL-6, IL- 7, IL-8, IL-10, TNF-α, and IFN-γ). Results. When analyzing the frequency of detection of Campylobacter sequence types in children with clinical intestinal infections, it was found that the profile of isolated isolates is most similar to those from countries of North America (USA and Canada), Northern Europe (Great Britain, Holland ) and Scandinavia (Denmark, Sweden, Finland). Identification of a pathogen with the flgE+, cdtA+, cdtC+ genotype was accompanied by a statistically significant increase in the level of IL-8 and a decrease in the content of IgA in the peripheral blood serum, which reflected the low efficiency of the immune response during infection with Campylobacter and predetermined the severe course of the infectious process during the disease.
导言。弯曲杆菌病是急性胃肠道感染的主要病因之一。弯曲杆菌病的严重程度和长期并发症的发生可能受病原体基因型的影响,而病原体的生物特性会影响免疫反应参数。本研究旨在确定弯曲杆菌病原体流行克隆的常见基因型,并调查免疫反应和疾病严重程度的特征。材料和方法。研究对象包括2019-2021年在联邦国家预算机构 "DNKCIB FMBA "诊所接受治疗的203名年龄在1个月至17岁之间的弯曲杆菌病患者。诊断采用聚合酶链式反应法进行确诊。患者样本也采用培养方法进行了分析。使用 QIAamp DNA Mini Kit 提取总 DNA。使用 ResFinder 程序对编码毒力因子的基因决定因子和 MLST 进行分型。在发病第 1 天和第 7 天对患者的免疫状态进行了评估。免疫学检查包括测定血清免疫球蛋白浓度(IgA、IgM、IgG)、C反应蛋白和细胞因子(IL-1β、IL-1、IL-2、IL-4、IL-5、IL-6、IL-7、IL-8、IL-10、TNF-α和IFN-γ)。结果在分析临床肠道感染儿童中弯曲杆菌序列类型的检测频率时发现,分离出的分离株的特征与北美(美国和加拿大)、北欧(英国、荷兰)和斯堪的纳维亚(丹麦、瑞典、芬兰)国家的分离株特征最为相似。在鉴定出具有 flgE+、cttA+、cttC+ 基因型的病原体的同时,外周血血清中的 IL-8 水平也出现了统计学意义上的显著升高,IgA 含量下降,这反映出在感染弯曲杆菌期间免疫反应的低效率,并预示着疾病期间感染过程的严重程度。
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引用次数: 0
Genomic diversity and analysis of resistance determinants of Salmonella enterica subspecies enterica serotype Kentucky isolated in Russia 俄罗斯分离的肠炎沙门氏菌亚种肯塔基血清型的基因组多样性和抗性决定因素分析
Pub Date : 2024-07-16 DOI: 10.36233/0372-9311-488
K. Kuleshov, A. S. Pavlova, Anna A. Kremleva, Anna E. Karpenko, Y. Mikhaylova, N.E. Krutova, Maria R. Lisitsyna, Kristina R. Popova, Olga A. Veselova, Alexandr T. Podkolzin, V. Akimkin
Introduction. Salmonella Kentucky sequence type ST198 is one of the epidemiologically significant non-typhoidal Salmonella clones worldwide and is characterized by the presence of highly resistant strains and the ability to adapt to different animal hosts and environmental conditions. The aim of this study was to analyze S. Kentucky strains isolated from various sources in Russia in terms of their phylogenetic position within the global diversity of the pathogen and their genetic characteristics. Materials and methods. We examined 55 strains of S. Kentucky by whole-genome sequencing, which were isolated from 2010 to 2022 from various sources (clinical strains, food, as well as from farm animals, feed and environmental samples). Whole genome sequencing was performed using Illumina platforms. Phylogenetic analysis based on nucleotide variation analysis included an additional 390 S. Kentucky strains. Results. Most of the Russian strains (n = 50) belonged to the ST198 sequence type, four strains were ST314 and one strain was ST152. Of the 50 Russian sequence-type ST198 strains, 44 belonged to the international monophyletic MDR lineage S. Kentucky ST198, and belonged to four separate sublineages, six strains occupying a basal position in relation to the MDR lineage. A total of 320 genes and mutations responsible for resistance to antimicrobial agents were identified. The most common were point mutations in the QRDR region. In most cases, Russian strains were characterized by the presence of variants of the SGI1-K genomic island. Moreover, the putative structure of SGI1 was correlated with the phylogenetic clustering of S. Kentucky sublineages. Conclusions. The results of the study made it possible to assess the population structure of Russian S. Kentucky ST198 strains on a global scale and determine the genetic determinants of antibiotic resistance, including the structure of the SGI1 genomic island.
导言。肯塔基沙门氏菌序列类型 ST198 是全球流行病学上重要的非伤寒沙门氏菌克隆之一,其特点是存在高抗性菌株,并能适应不同的动物宿主和环境条件。本研究的目的是分析从俄罗斯不同来源分离的肯塔基沙门氏菌菌株在全球病原体多样性中的系统发育位置及其遗传特征。材料和方法。我们通过全基因组测序检测了 55 株肯塔基杆菌菌株,这些菌株于 2010 年至 2022 年期间从不同来源(临床菌株、食品以及农场动物、饲料和环境样本)分离出来。全基因组测序使用 Illumina 平台进行。基于核苷酸变异分析的系统发育分析包括另外 390 株肯塔基州鼠疫菌株。结果大多数俄罗斯菌株(n = 50)属于 ST198 序列类型,4 株属于 ST314,1 株属于 ST152。在 50 株俄罗斯序列型 ST198 菌株中,44 株属于国际单系 MDR 系 S. Kentucky ST198,属于 4 个独立的亚系,其中 6 株相对于 MDR 系处于基础位置。共鉴定出 320 个导致对抗菌剂产生耐药性的基因和突变。最常见的是 QRDR 区域的点突变。在大多数情况下,俄罗斯菌株的特点是存在 SGI1-K 基因组岛变异。此外,SGI1 的推定结构与肯塔基州鼠亚系的系统发育聚类相关。结论研究结果有助于在全球范围内评估俄罗斯肯塔基沙雷菌 ST198 株系的种群结构,并确定抗生素耐药性的遗传决定因素,包括 SGI1 基因组岛的结构。
{"title":"Genomic diversity and analysis of resistance determinants of Salmonella enterica subspecies enterica serotype Kentucky isolated in Russia","authors":"K. Kuleshov, A. S. Pavlova, Anna A. Kremleva, Anna E. Karpenko, Y. Mikhaylova, N.E. Krutova, Maria R. Lisitsyna, Kristina R. Popova, Olga A. Veselova, Alexandr T. Podkolzin, V. Akimkin","doi":"10.36233/0372-9311-488","DOIUrl":"https://doi.org/10.36233/0372-9311-488","url":null,"abstract":"Introduction. Salmonella Kentucky sequence type ST198 is one of the epidemiologically significant non-typhoidal Salmonella clones worldwide and is characterized by the presence of highly resistant strains and the ability to adapt to different animal hosts and environmental conditions. \u0000The aim of this study was to analyze S. Kentucky strains isolated from various sources in Russia in terms of their phylogenetic position within the global diversity of the pathogen and their genetic characteristics. \u0000Materials and methods. We examined 55 strains of S. Kentucky by whole-genome sequencing, which were isolated from 2010 to 2022 from various sources (clinical strains, food, as well as from farm animals, feed and environmental samples). Whole genome sequencing was performed using Illumina platforms. Phylogenetic analysis based on nucleotide variation analysis included an additional 390 S. Kentucky strains. \u0000Results. Most of the Russian strains (n = 50) belonged to the ST198 sequence type, four strains were ST314 and one strain was ST152. Of the 50 Russian sequence-type ST198 strains, 44 belonged to the international monophyletic MDR lineage S. Kentucky ST198, and belonged to four separate sublineages, six strains occupying a basal position in relation to the MDR lineage. A total of 320 genes and mutations responsible for resistance to antimicrobial agents were identified. The most common were point mutations in the QRDR region. In most cases, Russian strains were characterized by the presence of variants of the SGI1-K genomic island. Moreover, the putative structure of SGI1 was correlated with the phylogenetic clustering of S. Kentucky sublineages. \u0000Conclusions. The results of the study made it possible to assess the population structure of Russian S. Kentucky ST198 strains on a global scale and determine the genetic determinants of antibiotic resistance, including the structure of the SGI1 genomic island.","PeriodicalId":508236,"journal":{"name":"Journal of microbiology, epidemiology and immunobiology","volume":"10 9","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-07-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141641193","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
The effect of carbohydrate adjuvants in the composition of the experimental hantavirus vaccine on the dynamics of neutralizing antibodies in the blood sera of guinea pigs 实验性汉坦病毒疫苗成分中的碳水化合物佐剂对豚鼠血清中中和抗体动态的影响
Pub Date : 2024-07-16 DOI: 10.36233/0372-9311-523
S. Kurashova, M. Egorova, A. Vetrova, M. Balovneva, Yulia V. Popova, R. Teodorovich, Anna A. Mironenko, P. G. Aparin, V. L. Lvov, Eugeny A. Tkachenko, T. Dzagurova
Introduction. Vaccination could be the most effective means of preventing hemorrhagic fever with renal syndrome (HFRS), which is one of the leading zoonoses and a major cause of natural disease in humans. Therefore, research to develop an effective vaccine and the search for new non-toxic and effective adjuvants that can enhance and prolong the immune response, reduce the antigen concentration and reduce the frequency of dose administration is an urgent task. The aim of the study was to determine the dependence of the immunogenic activity of an experimental hantavirus vaccine based on Puumala virus (HV) on the antigen dose both in native form and in combination with carbohydrate adjuvants. Materials and methods. The immunoadjuvant effect of Ac3-LPS S. sonnei and S. flexneri 1B and emulsions of nanoemulsified squalene Sepivac SWE (SWE) as part of HV at the optimal and minimal doses was investigated by induction of neutralising antibodies after 3-fold immunisation (day 0, 14, 182) of guinea pigs (Cavia porcellus). Differences in the immunogenic activity of HV were assessed in the neutralisation reaction by 50% suppression of focal units in Vero E6 cell cultures. Results. A statistically significant increase in neutralising antibodies levels was observed after immunization both with HV at the optimal dose and for its combination with adjuvants. Ac3-LPS S. sonnei and SWE showed the most pronounced immunoadjuvant effect at concentrations of 50 and 100 μg/dose, respectively. The immunoadjuvant effect of the combination of the hantavirus vaccine with Ac3-LPS S. sonnei and SWE contributed to both an enhanced immune response and its duration. The tenfold reduction of the antigen dose in the presence of SWE allows optimization of the immune response to the vaccine. Conclusion. The results of this study show the prospects of using Ac3-LPS S. sonnei and Sepivac SWE in the vaccine for the prevention of HFRS.
导言。疫苗接种可能是预防肾综合征出血热(HFRS)的最有效手段,肾综合征出血热是主要的人畜共患病之一,也是导致人类自然疾病的主要原因。因此,研究开发一种有效的疫苗,寻找能增强和延长免疫反应、降低抗原浓度和减少给药次数的无毒、有效的新型佐剂是一项紧迫的任务。本研究的目的是确定基于普马拉病毒(HV)的实验性汉坦病毒疫苗的免疫原活性对原形抗原剂量和与碳水化合物佐剂结合的抗原剂量的依赖性。材料和方法通过对豚鼠(Cavia porcellus)进行 3 次免疫(第 0、14、182 天)后诱导中和抗体,研究了作为 HV 一部分的 Ac3-LPS S. sonnei 和 S. flexneri 1B 以及纳米乳化角鲨烯 Sepivac SWE(SWE)乳剂在最佳和最小剂量下的免疫佐剂效应。通过在 Vero E6 细胞培养物中抑制 50%的病灶单位来评估 HV 在中和反应中的免疫原性差异。结果显示使用最佳剂量的 HV 及其与佐剂结合免疫后,中和抗体水平均有统计学意义上的显著提高。Ac3-LPS S. sonnei 和 SWE 的免疫佐剂效果最明显,浓度分别为 50 和 100 微克/剂量。汉坦病毒疫苗与 Ac3-LPS S. sonnei 和 SWE 的组合免疫佐剂效应有助于增强免疫反应及其持续时间。加入 SWE 后,抗原剂量减少了 10 倍,从而优化了疫苗的免疫反应。结论本研究结果表明,在疫苗中使用 Ac3-LPS S. sonnei 和 Sepivac SWE 有助于预防 HFRS。
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引用次数: 0
Influence of Yersinia pestis with different plasmid composition on the erythrocyte membrane in the blood of guinea pigs 不同质粒组成的鼠疫耶尔森菌对豚鼠血液中红血球膜的影响
Pub Date : 2024-07-16 DOI: 10.36233/0372-9311-471
S. Klyueva, S. Bugorkova, Pavel S. Erokhin, Anastasiya Yu. Goncharova, A. L. Kravtsov
Introduction. Based on data on the role of blood erythrocytes in the development and implementation of the vaccine and infectious processes in plague, it was of interest to evaluate changes in erythrocyte surface architecture from the position of searching for informative criteria for the preclinical evaluation of anti-plague vaccines. Aim — using atomic force microscopy to characterize the state of the blood erythrocyte membrane of guinea pigs in response to subcutaneous administration of the vaccine strain Yersinia pestis EV NIIEG and its isogenic derivatives. Materials and methods. For immunization of animals strain Y. pestis EV NIIEG (pYT+, pYV+, pYP+) and its isogenic derivatives Y. pestis KM216 (pYT–, pYV–, pYP+), Y. pestis KM217 (pYT–, pYV+, pYP–), Y. pestis KM218 (pYT–, pYV–, pYP–) were used. Analysis of the erythrocyte membrane was carried out using a Solver P47-PRO scanning probe microscope (NT-MDT, Russia). Results. The most pronounced changes in the surface architectonics of the membrane of guinea pig erythrocytes were established during the first three days of the formation of the immune response to the Y. pestis EV strain NIIEG and its isogenic variant Y. pestis KM217, in the genome of which the pYV plasmid is preserved, administered at a dose of 5 × 108 CFU. A significant increase (p 0.05) in the proportion of transformed cell forms (43.67 ± 3.63% and 37.83 ± 7.03% versus 4.08 ± 0.86% in the control group), root mean square roughness (319 ± 8 nm and 312 ± 7 nm versus 70 ± 6 nm in the control group), Young's modulus (125.73 ± 4.48 kPa and 113.8 ± 5.41 kPa versus 53.03 ± 1.47 kPa in the control group). By the 21st day, the value of these indicators decreased by an average of 2.7, 2.0 and 1.5 times, respectively, indicating restoration of the erythrocyte membrane. Conclusion. The dependence of the changes in the erythrocyte membrane and the rate of their restoration on the plasmid composition of Y. pestis strains has been established. The data obtained contribute to the understanding of the processes of interaction of Y. pestis with the erythrocyte membrane and can be used as additional characteristics in the development of new criteria for preclinical evaluation of plague candidate vaccines.
导言。根据有关血液红细胞在疫苗的开发和实施以及鼠疫感染过程中的作用的数据,从寻找抗鼠疫疫苗临床前评估的信息标准的角度出发,对红细胞表面结构的变化进行评估是很有意义的。目的--使用原子力显微镜描述豚鼠血红细胞膜对皮下注射鼠疫耶尔森菌 EV NIIEG 疫苗株及其同源衍生物的反应状态。材料和方法使用鼠疫耶尔森菌 EV NIIEG 株(pYT+, pYV+, pYP+)及其同源衍生物鼠疫耶尔森菌 KM216 株(pYT-, pYV-, pYP+)、鼠疫耶尔森菌 KM217 株(pYT-, pYV+, pYP-)、鼠疫耶尔森菌 KM218 株(pYT-, pYV-, pYP-)对动物进行免疫。使用 Solver P47-PRO 扫描探针显微镜(NT-MDT,俄罗斯)对红细胞膜进行了分析。结果显示豚鼠红细胞膜表面结构的最明显变化发生在对鼠疫伊蚊 EV 株 NIIEG 及其同源变种鼠疫伊蚊 KM217(其基因组中保留了 pYV 质粒)形成免疫反应的头三天,给药剂量为 5 × 108 CFU。转化细胞的比例(43.67 ± 3.63% 和 37.83 ± 7.03% 对对照组的 4.08 ± 0.86%)、均方根粗糙度(319 ± 8 nm 和 312 ± 7 nm 对对照组的 70 ± 6 nm)、杨氏模量(125.73 ± 4.48 kPa 和 113.8 ± 5.41 kPa 对对照组的 53.03 ± 1.47 kPa)均有明显增加(P 0.05)。到第 21 天,这些指标值分别平均下降了 2.7 倍、2.0 倍和 1.5 倍,表明红细胞膜已经恢复。结论红细胞膜的变化及其恢复速度与鼠疫伊蚊菌株的质粒组成有关。获得的数据有助于了解鼠疫酵母菌与红细胞膜的相互作用过程,并可作为开发鼠疫候选疫苗临床前评估新标准的补充特征。
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引用次数: 0
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Journal of microbiology, epidemiology and immunobiology
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