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The emerging threat of Japanese encephalitis in Australia: is it the time to worry? 澳大利亚新出现的日本脑炎威胁:是时候担心了吗?
IF 3 4区 医学 Q2 Medicine Pub Date : 2023-07-01 DOI: 10.1099/jmm.0.001738
Sameena Khan, Rajashri Patil, Nikunja Kumar Das, Sriram Kannuri, Sahjid Mukhida
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引用次数: 0
Vancomycin-sensitive Enterococcus faecium bacteraemia - hospital transmission and mortality in a Danish University Hospital. 万古霉素敏感屎肠球菌菌血症——丹麦大学医院的医院传播和死亡率。
IF 3 4区 医学 Q2 Medicine Pub Date : 2023-07-01 DOI: 10.1099/jmm.0.001731
Sanne Groenvall Kjaer Hansen, Louise Roer, Kasper Thystrup Karstensen, Silje Vermedal Hoegh, Frank Hansen, Kasper Klein, Flemming S Rosenvinge, Anette Holm, Marianne N Skov, Anette M Hammerum, Henrik Hasman

Introduction. The emergence of vancomycin-resistant Enterococcus faecium (VREfm) has left the vancomycin-sensitive E. faecium (VSEfm) strains almost unnoticed.Hypothesis. Molecular characteristics, hospital transmission patterns and clinical impact of VSEfm have changed, and VSEfm is a predictor of VREfm introduction.Aim. We wanted to do a molecular characterization of VSEfm to identify hospital transmissions and links between VSEfm and VREfm, and to investigate the demographics, treatment and impact on mortality of VSEfm bacteraemia.Methodology. VSEfm and VREfm blood culture isolates from Odense University Hospital, Denmark, from 2015 to 2019 were characterized using whole-genome sequencing and core-genome multilocus sequence typing (cgMLST). Clonal shifts and diversity of the VREfm isolates were compared to the VSEfm isolates. Hospital records were used for clinical data and transmission investigation of VSEfm cases.Results. Six-hundred and thirty VSEfm isolates from 599 patients belonged to 42 sequence types (STs) and 131 complex types (CTs) in several clusters. Multiple types were involved in putative transmission, occurring over the entire period. Twenty-seven VREfm bacteraemia cases were included. No correlation between the VSEfm and VREfm clones was identified. The 30 day mortality was 40 %, but only in 6.3 % of the cases, VSEfm bacteraemia was the likely cause of death.Conclusion. The molecular types of VSEfm bacteraemia isolates are changing and diverse. No direct correlation between VSEfm and the introduction of VREfm was found, but widespread hospital transmission indicates a presence of risk factors that could facilitate transmission of other micro-organisms as well. VSEfm bacteraemia is rarely the cause of death, indicating that 30 day mortality does not reflect the cause of death.

介绍。耐万古霉素屎肠球菌(VREfm)的出现使得对万古霉素敏感的屎肠球菌(VSEfm)几乎没有被注意到。VSEfm的分子特征、医院传播模式和临床影响已发生变化,VSEfm可作为VREfm引入的预测因子。我们希望对VSEfm进行分子表征,以确定VSEfm和VREfm之间的医院传播和联系,并调查VSEfm菌血症的人口统计学、治疗和对死亡率的影响。采用全基因组测序和核心基因组多位点序列分型(cgMLST)对2015 - 2019年来自丹麦欧登塞大学医院的VSEfm和VREfm血培养株进行鉴定。比较了VREfm菌株与VSEfm菌株的克隆转移和多样性。利用医院记录对VSEfm病例进行临床资料和传播调查。从599例患者中分离出的630株VSEfm在多个聚类中属于42个序列型(STs)和131个复杂型(ct)。假定的传播涉及多种类型,发生在整个时期。包括27例VREfm菌血症病例。VSEfm和VREfm克隆间无相关性。30天死亡率为40%,但仅6.3%的病例中,VSEfm菌血症是可能的死亡原因。VSEfm菌血症分离株的分子类型是变化的和多样的。没有发现VSEfm与引入VREfm之间的直接关联,但广泛的医院传播表明存在可能促进其他微生物传播的危险因素。VSEfm菌血症很少是死亡原因,这表明30天死亡率并不能反映死亡原因。
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引用次数: 1
Multiple mechanisms of linezolid resistance in Staphylococcus haemolyticus detected by whole-genome sequencing. 全基因组测序检测溶血葡萄球菌耐利奈唑胺多种机制。
IF 3 4区 医学 Q2 Medicine Pub Date : 2023-07-01 DOI: 10.1099/jmm.0.001737
Meerabai Manoharan, Ajit Ramesh Sawant, K Prashanth, Sujatha Sistla

Introduction. Linezolid is an effective therapeutic option for treating severe infections caused by multidrug-resistant Gram-positive organisms. Several mechanisms have been reported to be responsible for resistance to this antibiotic.Hypothesis or Gap Statement. Although several mechanisms of linezolid resistance have been reported in Staphylococcus haemolyticus, the prevalence and potential for horizontal transfer of resistance genes have not been fully characterized, particularly among S. haemolyticus isolates from India.Aim. To perform whole-genome sequencing (WGS) of linezolid-resistant S. haemolyticus isolates to characterize the resistance mechanisms.Methodology. WGS was performed for 16 linezolid-resistant S. haemolyticus isolates to check for the presence of cfr, optrA and poxtA genes and mutations in 23S rRNA and ribosomal proteins (L3, L4 and L22) that are possible mechanisms implicated in linezolid resistance. Sequence types were identified using MLST finder. The minimum inhibitory concentration (MIC) of linezolid was determined using the E-test method. Polymerase chain reaction (PCR) was carried out for the detection of the cfr gene.Results. The study documented three different mechanisms of linezolid resistance in S. haemolyticus. Thirteen of the 16 isolates were phenotypically resistant to linezolid, of which 12 were positive for the cfr gene. The G2603T mutation in 23S rRNA was found in the majority of the isolates (n=13). Ten isolates had the R138V mutation in L3 ribosomal protein. Twelve isolates with the cfr gene in combination with either G2603T or R138V mutations displayed extremely high MIC values. Surprisingly, three phenotypically sensitive isolates were found to be positive for the cfr gene but negative for other resistance mechanisms. Importantly, in almost half of the isolates the cfr gene was present on a plasmid. ST3 and ST1 were found to be the predominant sequence types.Conclusion. All phenotypically resistant isolates exhibited two or three linezolid resistance mechanisms. The cfr gene was found on plasmids in many isolates, demonstrating its potential for horizontal transfer to more pathogenic organisms.

介绍。利奈唑胺是治疗多重耐药革兰氏阳性菌引起的严重感染的有效治疗选择。据报道,几种机制导致对这种抗生素产生耐药性。假设或缺口陈述。虽然已经报道了溶血葡萄球菌耐利奈唑胺的几种机制,但尚未完全确定耐药基因的流行程度和水平转移的潜力,特别是在印度的溶血葡萄球菌分离株中。目的:对耐利奈唑胺的溶血链球菌进行全基因组测序,以确定其耐药机制。对16株耐利奈唑胺溶血链球菌进行WGS检测,以检测cfr、optrA和poxtA基因的存在,以及23S rRNA和核糖体蛋白(L3、L4和L22)的突变,这些突变可能与利奈唑胺耐药机制有关。利用MLST finder识别序列类型。采用e检验法测定利奈唑胺的最低抑菌浓度(MIC)。采用聚合酶链反应(PCR)检测cfr基因。该研究记录了溶血链球菌耐利奈唑胺的三种不同机制。16株菌株中有13株对利奈唑胺表型耐药,其中12株cfr基因阳性。在大多数分离株中发现了23S rRNA的G2603T突变(n=13)。10株分离株存在L3核糖体蛋白R138V突变。12株cfr基因与G2603T或R138V突变组合的分离株显示出极高的MIC值。令人惊讶的是,发现三个表型敏感的分离株对cfr基因呈阳性,但对其他耐药机制呈阴性。重要的是,在几乎一半的分离株中,cfr基因存在于质粒上。ST3和ST1为优势序列型。所有表型耐药菌株均表现出两种或三种利奈唑胺耐药机制。在许多分离株的质粒上发现了cfr基因,表明其具有向更多致病生物水平转移的潜力。
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引用次数: 1
Malaria vaccines to moderate the impact of antibiotic resistance in endemic countries. 疟疾疫苗,以减轻疟疾流行国家抗生素耐药性的影响。
IF 3 4区 医学 Q2 Medicine Pub Date : 2023-07-01 DOI: 10.1099/jmm.0.001734
Maria Khan, Saba Khan

Swift and widespread deployment of an efficient malaria vaccine in Pakistan, together with basic control and preventive measures, could significantly decrease the economic and healthcare burden caused by drug-resistant malaria. Moreover, the RTS,S/AS01 vaccine has provided a much needed breakthrough after decades of growth, as an innovative vaccine for malaria in phase 3 clinical trials and currently undergoing implementation studies. Vaccination is a potentially critical component of efforts to arrest the development and dissemination of antimicrobial resistance, although little is known about the impact vaccination may have within low- and-middle-income countries.

在巴基斯坦迅速和广泛地部署有效的疟疾疫苗,加上基本的控制和预防措施,可以大大减少耐药疟疾造成的经济和医疗负担。此外,RTS,S/AS01疫苗在经过几十年的发展之后提供了急需的突破,作为一种处于3期临床试验并正在进行实施研究的创新型疟疾疫苗。疫苗接种是遏制抗菌素耐药性发展和传播的一个潜在关键组成部分,尽管人们对疫苗接种在低收入和中等收入国家可能产生的影响知之甚少。
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引用次数: 0
Quantifying patient- and hospital-level antimicrobial resistance dynamics in Staphylococcus aureus from routinely collected data. 从常规收集的数据中量化金黄色葡萄球菌患者和医院水平的抗菌素耐药性动态。
IF 3 4区 医学 Q2 Medicine Pub Date : 2023-07-01 DOI: 10.1099/jmm.0.001724
Quentin Leclerc, Alastair Clements, Helen Dunn, James Hatcher, Jodi A Lindsay, Louis Grandjean, Gwenan M Knight

Introduction. Antimicrobial resistance (AMR) to all antibiotic classes has been found in the pathogen Staphylococcus aureus. The reported prevalence of these resistances varies, driven by within-host AMR evolution at the patient level, and between-host transmission at the hospital level. Without dense longitudinal sampling, pragmatic analysis of AMR dynamics at multiple levels using routine surveillance data is essential to inform control measures.Gap Statement. The value and limitations of routinely collected hospital data to gain insight into AMR dynamics at the hospital and individual levels simultaneously are unclear.Methodology. We explored S. aureus AMR diversity in 70 000 isolates from a UK paediatric hospital between 2000-2021, using electronic datasets containing multiple routinely collected isolates per patient with phenotypic antibiograms and information on hospitalization and antibiotic consumption.Results. At the hospital level, the proportion of isolates that were meticillin-resistant (MRSA) increased between 2014-2020 from 25-50 %, before sharply decreasing to 30%, likely due to a change in inpatient demographics. Temporal trends in the proportion of isolates resistant to different antibiotics were often correlated in MRSA, but independent in meticillin-susceptible S. aureus. Ciprofloxacin resistance in MRSA decreased from 70-40 % of tested isolates between 2007-2020, likely linked to a national policy to reduce fluoroquinolone usage in 2007. At the patient level, we identified frequent AMR diversity, with 4 % of patients ever positive for S. aureus simultaneously carrying, at some point, multiple isolates with different resistances. We detected changes over time in AMR diversity in 3 % of patients ever positive for S. aureus. These changes equally represented gain and loss of resistance.Conclusion. Within this routinely collected dataset, we found that 65 % of changes in resistance within a patient's S. aureus population could not be explained by antibiotic exposure or between-patient transmission of bacteria, suggesting that within-host evolution via frequent gain and loss of AMR genes may be responsible for these changing AMR profiles. Our study highlights the value of exploring existing routine surveillance data to determine underlying mechanisms of AMR. These insights may substantially improve our understanding of the importance of antibiotic exposure variation, and the success of single S. aureus clones.

介绍。在金黄色葡萄球菌中发现了对所有抗生素类的耐药性(AMR)。据报道,这些耐药性的流行程度各不相同,这是由患者水平的宿主内AMR演变和医院水平的宿主间传播驱动的。在没有密集的纵向抽样的情况下,利用常规监测数据对多个层面的抗菌素耐药性动态进行实际分析,对于告知控制措施至关重要。差距的声明。常规收集医院数据以同时了解医院和个人层面的抗菌素耐药性动态的价值和局限性尚不清楚。我们研究了2000-2021年间来自英国儿科医院的70000株金黄色葡萄球菌的AMR多样性,使用电子数据集,其中包含每位患者的多个常规收集的分离株,并提供抗生素表型图以及住院和抗生素使用信息。在医院层面,2014-2020年间,耐甲氧西林(MRSA)分离株的比例从25- 50%上升,然后急剧下降至30%,这可能是由于住院患者人口统计数据的变化。对不同抗生素耐药的分离株比例的时间趋势在MRSA中往往是相关的,但在甲氧西林敏感的金黄色葡萄球菌中是独立的。2007年至2020年间,MRSA对环丙沙星的耐药性从测试分离株的70- 40%下降,这可能与2007年减少氟喹诺酮类药物使用的国家政策有关。在患者水平上,我们发现了频繁的AMR多样性,4%的患者在金黄色葡萄球菌阳性的情况下同时携带多种具有不同耐药性的分离株。我们在3%的金黄色葡萄球菌阳性患者中检测到AMR多样性随时间的变化。这些变化同样代表了阻力的增加和减少。在这个常规收集的数据集中,我们发现患者金黄色葡萄球菌群体中65%的耐药性变化不能通过抗生素暴露或患者之间的细菌传播来解释,这表明通过频繁获得和丢失AMR基因的宿主内进化可能是这些变化的AMR谱的原因。我们的研究强调了探索现有常规监测数据以确定AMR潜在机制的价值。这些见解可能会大大提高我们对抗生素暴露变化的重要性的理解,以及单个金黄色葡萄球菌克隆的成功。
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引用次数: 0
Hidden bugs in a newly opened hospital: the distribution of skin microbiota among healthcare workers in a newly opened teaching hospital. 新开业医院中隐藏的细菌:新开业教学医院医护人员皮肤微生物群分布
IF 3 4区 医学 Q2 Medicine Pub Date : 2023-07-01 DOI: 10.1099/jmm.0.001732
Manggaiyakarasi Kamala Nathan, Azmiza Syawani Jasni, Siti Zulaikha Zakariah, Tengku Zetty Maztura Tengku Jamaluddin, Muhammad Mohd Isa, Rosni Ibrahim

Background. Skin is a reservoir for millions of micro-organisms, all of which make up the skin microbiota. Hospitals have been identified as a favourable environment for transmitting micro-organisms and thus, it is important to know the distribution of skin microbiota among healthcare workers (HCWs), as such findings may provide baseline information for the distribution of skin microbiota in hospitals.Hypothesis. There is no significant association between the factors (age, gender, type of skin microenvironment, hand hygiene practices, usage of skin care products, current healthcare practices and previous workplace) and the distribution of the skin microbiota among HCWs.Aim. The study aims to identify type of skin microbiota and associated factors (age, gender, type of skin microenvironment, hand hygiene practices, use of skincare products, current healthcare practice, and previous workplace) that influence the growth of skin microbiota.Method. About 102 bacterial isolates were obtained from the skin of 63 healthcare workers in a newly opened teaching hospital, namely Hospital Pengajar Universiti Putra Malaysia (HPUPM). All isolated bacteria were subjected to phenotypic identification according to standard microbiological procedures.Results. The most common isolated skin microbiota were Gram-positive bacteria (84.3%), followed by Gram-negative bacteria (15.7%). A Chi-square test of independence was used to analyse the above factors and there was a significant association between the type of skin microenvironment and the distribution of skin microbiota (P=0.03) (type of skin microenvironment influences the distribution of skin microbiota).Conclusion. Coagulase-negative Staphylococcus spp. was the most common bacteria isolated from the skin of the healthcare workers. Even though coagulase-negative staphylococci (CoNS) are low pathogenic bacteria, but it may cause serious infection in high risk group of patients. Therefore, it is important to emphasize on the good hand hygiene practices and implement strict infection control measures to minimize the risk of HAI in newly opened hospitals.

背景。皮肤是数百万微生物的储存库,所有这些微生物组成了皮肤微生物群。医院已被确定为微生物传播的有利环境,因此,了解医护人员(HCWs)皮肤微生物群的分布是很重要的,因为这些发现可能为医院皮肤微生物群的分布提供基线信息。这些因素(年龄、性别、皮肤微环境类型、手部卫生习惯、护肤品使用情况、目前的保健习惯和以前的工作场所)与医护人员皮肤微生物群分布之间没有显著关联。该研究旨在确定影响皮肤微生物群生长的皮肤微生物群类型和相关因素(年龄、性别、皮肤微环境类型、手部卫生习惯、护肤品的使用、当前的医疗保健实践和以前的工作场所)。在新开设的教学医院,即马来西亚蓬加尔大学医院(HPUPM),从63名卫生保健工作者的皮肤中分离出约102株细菌。所有分离的细菌均按照标准微生物学程序进行表型鉴定。最常见的皮肤分离菌群为革兰氏阳性菌(84.3%),其次为革兰氏阴性菌(15.7%)。采用卡方独立性检验对上述因素进行分析,皮肤微环境类型与皮肤微生物群分布之间存在显著相关性(P=0.03)(皮肤微环境类型影响皮肤微生物群分布)。从医护人员皮肤中分离到的最常见细菌为凝固酶阴性葡萄球菌。虽然凝固酶阴性葡萄球菌(con)是一种低致病性细菌,但在高危人群中可能引起严重感染。因此,应重视良好的手卫生习惯,实施严格的感染控制措施,以尽量减少新开业医院发生HAI的风险。
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引用次数: 0
JMM Profile: West Nile virus. JMM简介:西尼罗河病毒。
IF 3 4区 医学 Q2 Medicine Pub Date : 2023-07-01 DOI: 10.1099/jmm.0.001730
Sanam Sewgobind, Fiona McCracken, Mirjam Schilling

West Nile virus (WNV) is a positive-sense single-stranded RNA virus belonging to the Flaviviridae family and is maintained in an enzootic cycle between avian hosts and mosquito vectors. Humans, horses and other mammals are susceptible to infection but are dead-end hosts due to a low viraemia. The disease can manifest itself in a variety of clinical signs and symptoms in people and horses from mild fever to severe encephalitis and morbidity. There are no vaccines licensed for human protection, but parts of Europe, North America, Africa and Australia have vaccines commercially available for horses.

西尼罗病毒(WNV)是黄病毒科的一种正义单链RNA病毒,在鸟类宿主和蚊子媒介之间保持着一种地方流行循环。人类、马和其他哺乳动物易受感染,但由于病毒血症低,它们是终端宿主。该病可在人和马中表现为各种临床体征和症状,从轻度发热到严重脑炎和发病率。目前还没有批准用于人类保护的疫苗,但欧洲、北美、非洲和澳大利亚的部分地区有用于马的市售疫苗。
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引用次数: 1
Serological status for TORCH in women of childbearing age: a decade-long surveillance (2012-2022) in Italy. 育龄妇女的 TORCH 血清学状况:意大利长达十年的监测(2012-2022 年)。
IF 2.4 4区 医学 Q3 MICROBIOLOGY Pub Date : 2023-07-01 DOI: 10.1099/jmm.0.001733
Emilia Palazzotto, Floriana Bonura, Cinzia Calà, Giuseppina Capra, Daniela Pistoia, Donatella Mangione, Chiara Mascarella, Giuseppe Minì, Marco Enea, Giovanni M Giammanco, Donatella Ferraro, Simona De Grazia

Introduction. Serological screening and seroprevalence data for TORCH infections represent a key instrument to estimate immunity and vaccination levels and exposure rates to prevent and treat TORCH congenital infections.Hypothesis. Serology allows us to identify women susceptible to primary infection.Aim. Assess the prevalence of women at risk of primary infections by TORCH pathogens in Palermo, Sicily, Italy, in the decade 2012-2022.Methodology. A retrospective study was performed to evaluate the serological status (IgG and/or IgM) of 2359 women of childbearing age (WCBA), ranging from 16 to 46 years, attending the AOUP 'P. Giaccone' University Hospital of Palermo.Results. The results showed an overall prevalence of anti-TORCH IgG of 90.5 % for herpesvirus (HSV), 81.2 % for rubella virus (RV), 72.1 % for cytomegalovirus (CMV), 20.9 % for Toxoplasma gondii (TOX) and 4.8 % for Treponema pallidum (TP). IgM positivity was 16.9 % for HSV2, 10.3 % for TOX, 4 % for CMV and, 2 % for RV. A recent/active infection by TP was confirmed in 28.3 % of the seropositive women. Our results indicate that only a small percentage of WCBA were subjected to a comprehensive TORCH serological screening, while most WCBA were only tested for a single pathogen. In addition, no significant differences were found in terms of the overall TORCH IgG seroprevalence among different age groups (P>0.05).Conclusion. Identifying WCBA at risk of exposure during pregnancy allows us to prevent and reduce possible congenital infections, providing detailed guidelines and instructions. The results of this study showed that in Italy the risk of acquiring a primary infection by a TORCH agent is still high, therefore effective prevention strategies, including serological screening, should be implemented.

导言。针对 TORCH 感染的血清学筛查和血清流行率数据是估算免疫和疫苗接种水平以及接触率的重要工具,以预防和治疗 TORCH 先天性感染。血清学使我们能够确定易受原发性感染的妇女。评估2012-2022年期间意大利西西里岛巴勒莫地区易受TORCH病原体原发性感染的妇女的发病率。我们开展了一项回顾性研究,以评估在 AOUP 'P. Giaccone'大学医院就诊的 2359 名 16 至 46 岁育龄妇女(WCBA)的血清状况(IgG 和/或 IgM)。Giaccone' University Hospital of Palermo)就诊的 2359 名育龄妇女的血清状况(IgG 和/或 IgM)。结果显示,疱疹病毒(HSV)、风疹病毒(RV)、巨细胞病毒(CMV)、弓形虫(TOX)和苍白曲霉(TP)的抗 TORCH IgG 阳性率分别为 90.5%、81.2%、72.1%、20.9% 和 4.8%。HSV2 的 IgM 阳性率为 16.9%,TOX 阳性率为 10.3%,CMV 阳性率为 4%,RV 阳性率为 2%。28.3%的血清反应呈阳性的妇女被证实最近/正在感染 TP。我们的结果表明,只有一小部分 WCBA 接受了全面的 TORCH 血清学筛查,而大多数 WCBA 只接受了单一病原体的检测。此外,不同年龄组的 TORCH IgG 血清阳性率也无明显差异(P>0.05)。通过识别孕期有接触风险的 WCBA,我们可以预防和减少可能的先天性感染,并提供详细的指导和说明。这项研究的结果表明,在意大利,感染 TORCH 病原体的风险仍然很高,因此应实施有效的预防策略,包括血清学筛查。
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引用次数: 0
MARGINAL NOTES, March 2023 - point of care? 边际笔记,2023年3月-护理点?
IF 3 4区 医学 Q2 Medicine Pub Date : 2023-07-01 DOI: 10.1099/jmm.0.001740
T J J Inglis
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引用次数: 0
Novel neutralizing mouse-human chimeric monoclonal antibodies against the SARS-CoV-2 receptor binding domain. 针对SARS-CoV-2受体结合域的新型中和小鼠-人嵌合单克隆抗体。
IF 3 4区 医学 Q2 Medicine Pub Date : 2023-06-01 DOI: 10.1099/jmm.0.001728
Ahmad Ghorbani, Faezeh Maghsood, Hamidreza Yadegari, Tannaz Bahadori, Amir-Hassan Zarnani, Seyed Alireza Razavi, Mahmood Jeddi-Tehrani, Mohammad Mehdi Amiri, Fazel Shokri

Introduction. Neutralizing antibodies have been widely used for the prophylaxis and treatment of COVID-19.Hypothesis. The major target for these neutralizing antibodies is the receptor-binding domain (RBD) of the viral spike protein.Aim. In the present study, we developed and characterized three neutralizing chimeric mouse-human mAbs for potential therapeutic purposes.Methodology. Light and heavy chain variable region genes of three mouse mAbs (m4E8, m3B6, and m1D1) were amplified and ligated to human Cγ1 and Cκ constant region genes by PCR. After cloning into a dual promoter mammalian expression vector, the final constructs were transiently expressed in DG-44 cells and the purified chimeric antibodies were characterized by ELISA and Western blotting. The neutralizing potency of the chimeric mAbs was determined by three different virus neutralization tests including sVNT, pVNT, and cVNT.Results. All three recombinant chimeric mAbs display human constant regions and are able to specifically bind to the RBD of SARS-CoV-2 with affinities comparable to the parental mAbs. Western blot analysis showed similar epitope specificity profiles for both the chimeric and the parental mouse mAbs. The results of virus neutralization tests (sVNT, pVNT, and cVNT) indicate that c4E8 had the most potent neutralizing activity with IC50 values of 1.772, 0.009, and 0.01 µg ml-1, respectively. All chimeric and mouse mAbs displayed a similar pattern of reactivity with the spike protein of the SARS-CoV-2 variants of concern (VOC) tested, including alpha, delta, and wild-type.Conclusion. The chimeric mAbs displayed neutralizing potency similar to the parental mouse mAbs and are potentially valuable tools for disease control.

介绍。中和抗体已广泛应用于covid -19的预防和治疗。这些中和抗体的主要靶点是病毒刺突蛋白的受体结合域(RBD)。在本研究中,我们开发并鉴定了三种具有潜在治疗目的的中和嵌合小鼠-人单克隆抗体。扩增小鼠单克隆抗体(m4E8、m3B6和m1D1)的轻链和重链可变区基因,并通过PCR与人c - γ1和Cκ恒定区基因连接。克隆到双启动子表达载体后,最终构建物在DG-44细胞中瞬间表达,纯化的嵌合抗体通过ELISA和Western blotting进行鉴定。通过三种不同的病毒中和试验,包括sVNT、pVNT和cvnt,来确定嵌合单抗的中和效力。所有三种重组嵌合单克隆抗体均显示人类恒定区,并能够特异性结合SARS-CoV-2的RBD,其亲和力与亲本单克隆抗体相当。Western blot分析显示嵌合和亲本小鼠单克隆抗体的表位特异性相似。病毒中和试验(sVNT、pVNT和cVNT)结果表明,c4E8具有最强的中和活性,IC50值分别为1.772、0.009和0.01µg ml-1。所有嵌合抗体和小鼠单抗均与SARS-CoV-2相关变异(VOC)的刺突蛋白(spike protein)表现出相似的反应模式,包括α型、δ型和野生型。嵌合单克隆抗体显示出与亲代小鼠单克隆抗体相似的中和效力,是潜在的有价值的疾病控制工具。
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