Serological survey

血清学调查
  • 文章类型: Journal Article
    巴泰病毒(BATV),一种由蚊子传播的人畜共患病原体,感染脊椎动物,包括牲畜,鸟,和人类。尽管在云南省已经检测到并隔离了BATV的蚊子,中国,没有牲畜感染的报告。因此,我们对红河州2021年至2022年春秋采集的牛、山羊血清中的BATV进行了分子和血清学调查,云南省,在中越边境。
    我们使用间接酶联免疫吸附测定和逆转录实时PCR(RT-qPCR)对929头牛和973头山羊血清样品进行了测试。
    在262/929(28.2%)牛和263/973(27.0%)山羊血清样品中检测到BATV抗体。RT-qPCR未检测到BATVRNA。
    芦溪县牛、山羊BATV血清抗体阳性率高于其他地区,秋季也比春季高,这可能与气候有关,温度,和蚊子密度。尽管我们的发现表明该地区的牲畜中存在BATV感染,RT-qPCR未检测到BATVRNA。因此,秋季应加强牛和山羊的BATV监测,应扩大宿主监测范围,明确BATV感染的宿主和媒介。
    UNASSIGNED: Batai virus (BATV), a zoonotic pathogen transmitted by mosquitoes, infects vertebrates, including livestock, birds, and humans. Although BATV has been detected and isolated in mosquitoes in Yunnan Province, China, there have been no reports of livestock infection. Thus, we conducted a molecular and serological investigation of BATV in cattle and goat sera collected in spring and autumn from 2021 to 2022 in Honghe Prefecture, Yunnan Province, on the China-Vietnam border.
    UNASSIGNED: We used indirect enzyme-linked immunosorbent assays and reverse transcription real-time PCR (RT-qPCR) to test 929 cattle and 973 goat serum samples.
    UNASSIGNED: BATV antibodies were detected in 262/929 (28.2%) cattle and 263/973 (27.0%) goat serum samples. RT-qPCR did not detect BATV RNA.
    UNASSIGNED: The positive rate of BATV serum antibodies in cattle and goats in Luxi County was higher compared with other areas, and it was also higher in autumn compared with spring, which may be related to climate, temperature, and mosquito density. Although our findings indicated the presence of BATV infection in livestock in the region, RT-qPCR did not detect BATV RNA. Therefore, BATV monitoring in cattle and goats should be heightened in autumn, and the scope of host monitoring should be expanded to clarify the hosts and vectors of BATV infection.
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  • 文章类型: Journal Article
    COVID-19,由严重急性呼吸综合征冠状病毒2(SARS-CoV-2)引起,可以刺激严重肺部受累的全身炎症反应,多系统功能障碍,和死亡在某些情况下。免疫抑制治疗已被提议用于COVID-19患者的管理,但是这些会带来预先存在的感染爆发的风险。在免疫功能低下的个体中,圆形线虫病可能变得严重或致命。这项横断面研究确定了从2021年1月至2022年1月在泰国一家三级医院从SARS-CoV-2感染者收集的血清中抗-圆线虫IgG抗体的患病率。这项调查是使用快速免疫色谱测试(ICT)试剂盒进行的,该试剂盒基于已知具有IgG免疫反应性的圆形圆线虫的重组抗原。发现抗圆线虫IgG抗体的患病率很高。在297名SARS-CoV-2感染患者中,117人(39.4%,根据ICT试剂盒,95%CI33.8-45.2%)为胸骨链球菌阳性。在线虫病流行的地区,我们建议在即将接受免疫抑制治疗的重症COVID-19患者中,使用这种即时医疗ICT试剂盒进行常规快速筛查.应及时进行驱虫治疗,以预防高危患者严重的全身性圆线虫病。
    COVID-19, caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), can stimulate a systemic inflammatory response with severe lung involvement, multisystem dysfunction, and death in some cases. Immunosuppressive treatments have been proposed for management of COVID-19 patients, but these bring the risk of flare-up of pre-existing infections. Strongyloidiasis can become severe or fatal in immunocompromised individuals. This cross-sectional study determined the prevalence of anti-Strongyloides IgG antibody in sera collected from SARS-CoV-2 infected persons in a tertiary-care Thai hospital from January 2021 to January 2022. The survey was conducted using a rapid immunochromatographic test (ICT) kit based on a recombinant antigen of Strongyloides stercoralis known to be IgG-immunoreactive. High prevalence of anti-Strongyloides IgG antibody was found. Out of 297 SARS-CoV-2-infected patients 117 (39.4 %, 95 % CI 33.8-45.2 %) were positive for S. stercoralis according to the ICT kit. In areas where strongyloidiasis is endemic, we suggest using this point-of-care ICT kit for routine rapid screening in seriously ill COVID-19 patients who will be subjected to immunosuppressive treatment. Prompt anthelminthic treatment should be administered to prevent serious systemic strongyloidiasis in at-risk patients.
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  • 文章类型: Journal Article
    随着2019年冠状病毒病(COVID-19)大流行期间学校的重新开放,必须了解学生和教育专业人员在严重急性呼吸道综合征冠状病毒2(SARS-CoV-2)传播中的作用.在本文中,我们确定了CampoGrande学校社区中SARS-CoV-2抗核衣壳抗体的血清阳性率,南马托格罗索州(巴西)的首府和人口最多的城市,并评估了其与性别的关系,学校水平,和学校类型。
    这项调查是使用免疫生物技术研究所的TRDPP®COVID-19免疫球蛋白M/免疫球蛋白G(IgM/IgG)试剂盒在CampoGrande城市地区的20所公立和私立学校中进行的(Bio-Manguinhos,里约热内卢,巴西)。测试分三个阶段进行:2021年10月至12月;2022年3月至7月;2022年8月至11月。参与者是小学或中学的6-17岁学生以及不同年龄和角色的专业人员。
    在第一个测试期间,162名参与者的IgM和/或IgG抗核衣壳SARS-CoV-2抗体呈血清阳性,使用贝叶斯多水平回归估计血清阳性率为19.6%。在第二阶段,251名参与者血清呈阳性(估计血清阳性率,34.6%),而在第三阶段,393名参与者血清转换(估计血清阳性率,56.7%)。2022年,与2021年相比,血清转化率有所上升。这三个时期最常见的急性表现是发烧,头痛,喉咙痛,流鼻涕.就人口统计而言,两性之间没有明显的血清阳性,尽管女性约占研究人群的70%。学生和学校工作人员之间也没有差异。
    这些结果使我们能够通过对SARS-CoV-2病毒的免疫来评估学校社区传播的程度,除了提供有关儿童COVID-19症状的信息外,青少年,和成年人。
    With the reopening of schools during the coronavirus disease 2019 (COVID-19) pandemic, it was imperative to understand the role of students and education professionals in the spread of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). In this paper, we determined the seroprevalence of the SARS-CoV-2 anti-nucleocapsid antibodies in the school community in Campo Grande, the capital and most populous city of the state of Mato Grosso do Sul (Brazil) and evaluated its association with sex, school level, and school type.
    The survey was carried out in 20 public and private schools in the urban region of Campo Grande using the TR DPP® COVID-19 immunoglobulin M/immunoglobulin G (IgM/IgG) kit from the Immunobiological Technology Institute (Bio-Manguinhos, Rio de Janeiro, Brazil). Testing was carried out in three periods: from October to December 2021; from March to July 2022; and from August to November 2022. The participants were students aged 6-17 years enrolled in primary or secondary schools and professionals of different ages and roles.
    During the first testing period, 162 participants were seropositive for the IgM and/or IgG anti-nucleocapsid SARS-CoV-2 antibodies, with an estimated seroprevalence of 19.6% using Bayesian multilevel regression. In the second period, 251 participants were seropositive (estimated seroprevalence, 34.6%), while in the third period, 393 participants were seroconverted (estimated seroprevalence, 56.7%). In 2022, there was an increase in the seroconversion rate compared to that in 2021. The most frequently described acute manifestations in the three periods were fever, headache, sore throat, and runny nose. In terms of the demographic profile, there was no predominance of seropositivity between the sexes, although women represented approximately 70% of the study population. There were also no differences between students and school staff.
    The results made it possible to evaluate the extent of SARS-CoV-2 transmission in the school community through immunity developed against the virus, in addition to providing information about COVID-19 symptoms in children, adolescents, and adults.
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  • 文章类型: Journal Article
    SARS-CoV-2的血清学调查在COVID-19大流行的早期用于评估流行病学情景。在卡斯卡伊斯市(葡萄牙),血清学检测与全面的社会人口统计学相结合,在2020年5月至2021年初期间,向居民提供了临床和行为问卷。在这项研究中,我们分析了与坚持这项市政倡议相关的因素,以及与血清阳性相关的社会人口统计学特征和慢性疾病的临床相关性。我们的目标是为今后的大流行防备工作提供相关信息。
    这是一项非概率抽样的横断面研究。居住在卡斯卡伊斯市的公民自愿前往采血中心参加血清学调查。参与者的比例,按社会人口统计学变量分层,与人口普查比例进行比较,以确定对调查依从性较低的群体。使用单变量和多变量逻辑回归来识别社会人口学,与血清阳性相关的临床和行为因素。
    从2020年5月到2021年2月,有19,608名参与者(占卡斯卡伊斯居民的9.2%)被纳入研究。根据与人口普查数据的比较,对这项调查依从性较低的群体是男性,最年轻和最年长的年龄组,受教育程度较低和失业/不活跃的个人。反应性(阳性)血清学测试的重要预测因素是年龄较小,被雇用或学生,住在更大的家庭。患有慢性疾病的个体通常表现出较低的血清阳性率。
    对这项自愿研究依从性低的群体,以及被确定为更有病毒传播风险的社会经济环境,可能会成为未来大流行形势的目标。我们还发现患有慢性疾病的人,认为患严重疾病的风险更高,采取了限制感染率的保护行为,揭示了预防措施的健康教育对这些患者是有效的。
    Serological surveys for SARS-CoV-2 were used early in the COVID-19 pandemic to assess epidemiological scenarios. In the municipality of Cascais (Portugal), serological testing combined with a comprehensive socio-demographic, clinical and behavioral questionnaire was offered to residents between May 2020 and beginning of 2021. In this study, we analyze the factors associated with adherence to this municipal initiative, as well as the sociodemographic profile and chronic diseases clinical correlates associated to seropositivity. We aim to contribute with relevant information for future pandemic preparedness efforts.
    This was a cross-sectional study with non-probabilistic sampling. Citizens residing in Cascais Municipality went voluntarily to blood collection centers to participate in the serological survey. The proportion of participants, stratified by socio-demographic variables, was compared to the census proportions to identify the groups with lower levels of adherence to the survey. Univariate and multivariate logistic regression were used to identify socio-demographic, clinical and behavioral factors associated with seropositivity.
    From May 2020 to February 2021, 19,608 participants (9.2% of the residents of Cascais) were included in the study. Based on the comparison to census data, groups with lower adherence to this survey were men, the youngest and the oldest age groups, individuals with lower levels of education and unemployed/inactive. Significant predictors of a reactive (positive) serological test were younger age, being employed or a student, and living in larger households. Individuals with chronic diseases generally showed lower seroprevalence.
    The groups with low adherence to this voluntary study, as well as the socio-economic contexts identified as more at risk of viral transmission, may be targeted in future pandemic situations. We also found that the individuals with chronic diseases, perceiving higher risk of serious illness, adopted protective behaviors that limited infection rates, revealing that health education on preventive measures was effective for these patients.
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  • 文章类型: Journal Article
    蜱传脑炎仅限于意大利东北部。我们在伦巴第报道,西北部一个人口众多的地区,一种表现出有多个病毒阳性蜱的蜱传脑炎病毒临床症状的羚羊,以及一个有症状的人。Further,我们显示了该地区病毒循环的血清学证据。
    Tick-borne encephalitis was limited to northeast portions of Italy. We report in Lombardy, a populous region in the northwest, a chamois displaying clinical signs of tickborne encephalitis virus that had multiple virus-positive ticks attached, as well as a symptomatic man. Further, we show serologic evidence of viral circulation in the area.
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  • 文章类型: Journal Article
    牛布鲁氏菌病是一种世界性的人畜共患传染病。根据世界动物卫生信息系统的报告,厄瓜多尔在过去几年中在大陆地区爆发了越来越多的牛布鲁氏菌病(2018年为756起,2021年为964起)。对生产者造成经济损失,对公众健康造成风险。在2018年5月至6月之间进行了一项横断面研究,以调查牛布鲁氏菌病的血清阳性率及其相关风险或保护因素。这项分层随机研究是在厄瓜多尔大陆23个省的290个牛群中进行的,总共有3737头24个月或以上的奶牛。使用竞争性ELISA检测布鲁氏菌抗体。同时,我们进行了流行病学调查,以评估布鲁氏菌病的风险或保护因素。牛群水平的牛布鲁氏菌病的明显患病率为21.3%(95%CI:16.8-26.6)和6.2%(95%CI:5.5-7)。进行了单变量和多变量逻辑回归分析,以确定与牛布氏杆菌病相关的潜在因素之间的关系。多变量分析后确定的危险因素是每群公顷>70公顷(OR=2.73;95%CI:1.18-6.32)和每只动物的分娩数量(OR≥1.8且p值≤0.047的两个或更多)。相反,保护因素是该地区(农场位于东部地区)和没有报告的临床体征.此外,在广泛生产占主导地位的牛群中,农民的知识水平很低,农场生物安全水平较低。这些结果可以指导当局管理已识别的风险因素,了解厄瓜多尔目前的流行病学情况,改善牛布鲁氏菌病控制计划和食品安全,以及增加单一健康的方法。
    Bovine brucellosis is a worldwide zoonotic contagious disease. According to World Animal Health Information System reports Ecuador has presented an increasing number of bovine brucellosis outbreaks in the continental territory over the past years (756 in 2018 versus 964 in 2021), generating economic losses for producers and causing a risk to public health. A cross-sectional study was conducted to investigate the seroprevalence of bovine brucellosis and associated risk or protective factors between May and June 2018. This stratified random study was implemented in 290 cattle herds located in the 23 provinces of continental Ecuador, which represents a total of 3737 cows aged 24 months or older. A competitive ELISA was used to detect Brucella antibodies. Simultaneously, an epidemiological survey was implemented to assess the brucellosis risk or protective factors. The apparent prevalence of bovine brucellosis at the herd level was 21.3% (95% CI: 16.8-26.6) and 6.2% (95% CI: 5.5-7) at the animal level. Univariate and multivariate logistic regression analyses were performed to determine the relationship between the potential factors associated with the presence of bovine brucellosis. The risk factors identified after multivariate analysis were a surface in ha per herd > 70 ha (OR = 2.73; 95% CI: 1.18-6.32) and the number of parturitions per animal (two or more with OR ≥ 1.8 and p-value ≤ 0.047). On the contrary, the protective factors were the region (farms located in the eastern region) and the absence of reported clinical signs. In addition, in herds where extensive production predominates, farmers have a low level of knowledge, and the farm biosecurity level is low. These results can guide the authorities in managing the risk factors identified, understanding the current epidemiological situation in Ecuador, improving the bovine brucellosis control program and food safety, as well as increase the one-health approach.
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  • 文章类型: Journal Article
    SenecavirusA(SVA)是一种新兴的病原体,对中国的养猪业产生负面影响。受影响的动物表现出与其他囊泡疾病难以区分的囊泡病变。迄今为止,在中国,没有可用于控制SVA感染的商业疫苗。在这项研究中,重组SVA3AB,2C,3C,3D,通过使用原核表达系统表达L和VP1蛋白。SVA抗体与SVA接种的猪血清的存在和水平的动力学表明3AB具有最佳的抗原性。用3AB蛋白开发了间接酶联免疫吸附测定(ELISA),灵敏度为91.3%,与PRRSV血清抗体无交叉反应,CSFV,PRV,PCV2或O型FMDV。鉴于这种方法的高灵敏度和特异性,我们进行了一项为期9年(2014-2022年)的回顾性和前瞻性血清学研究,以确定华东地区SVA的流行病学特征和动态.尽管SVA血清阳性从2016年(98.85%)到2022年(62.40%)显著下降,SVA传输在中国仍在继续。因此,基于SVA3AB的间接ELISA具有良好的敏感性和特异性,适用于病毒检测,现场监测和流行病学研究。
    Senecavirus A (SVA) is an emerging pathogen that negatively affects the pig industry in China. Affected animals present vesicular lesions which are indistinguishable from other vesicular diseases. To date, there is no commercial vaccine that can be used to control SVA infection in China. In this study, recombinant SVA 3AB, 2C, 3C, 3D, L and VP1 proteins are expressed by using a prokaryotic expression system. The kinetics of the presence and levels of SVA antibodies with SVA-inoculated pig serum show that 3AB has the best antigenicity. An indirect enzyme-linked immunosorbent assay (ELISA) is developed with the 3AB protein, exhibiting a sensitivity of 91.3% and no cross-reaction with serum antibodies against PRRSV, CSFV, PRV, PCV2 or O-type FMDV. Given the high sensitivity and specificity of this approach, a nine-year (2014-2022) retrospective and prospective serological study is conducted to determine the epidemiological profile and dynamics of SVA in East China. Although SVA seropositivity declined markedly from 2016 (98.85%) to 2022 (62.40%), SVA transmission continues in China. Consequently, the SVA 3AB-based indirect ELISA has good sensitivity and specificity and is suitable for viral detection, field surveillance and epidemiological studies.
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  • 文章类型: Journal Article
    猪链球菌是一种重要的人畜共患病原体,来自病态和隐性载体猪,对畜牧业生产和公共卫生构成严重威胁。它通常导致猪之间的水平传播。这种疾病的发病率和死亡率都很高。人类感染是通过与病猪直接或间接接触而引起的。中国的两次大规模疫情是由于猪场爆发猪链球菌,传播到人类感染;因此,检测猪群中的猪链球菌至关重要。目前,市售猪链球菌ELISA2型试剂盒只能检测单一血清型,相互作用反应的高概率,以及使用灭活的猪链球菌作为抗原时的生物安全风险。磷酸-3-甘油醛脱氢酶(GAPDH),muramidase-releasedprotein(MRP),二氢硫酰胺脱氢酶(DLDH)是重要的猪链球菌2型,猪链球菌7型和猪链球菌9型保护性抗原。本研究纯化了GMD蛋白(GAPDH的B细胞优势表位,MRP,和DLDH抗原),并使用多种不同抗原的优势表位的多种组合作为包被抗原,提高了间接ELISA实验的敏感性和安全性。建立了检测猪链球菌抗体的间接ELISA方法(GMD-ELISA)。使用棋盘滴定优化抗原-抗体应答。使用ELISA检测肠沙门氏菌的结果(S.enterica),大肠杆菌(E.大肠杆菌),金黄色葡萄球菌(SA),和化脓性链球菌(S.化脓性)都是阴性的,说明该方法具有较强的特异性。当猪链球菌阳性血清的稀释比例达到1:6,400时,结果仍然为阳性,因此表明该方法具有较高的灵敏度。间接ELISA的重复性测定结果表明,测定内变异系数和测定间变异系数均小于10%,表明该方法具有良好的重复性。我们调查了在华东地区收集的167份血清样本中猪链球菌的血清阳性率,33.5%的样本抗猪链球菌抗体呈阳性,这表明中国东部猪场猪链球菌的患病率很高。新型GMD-ELISA是一种方便的,敏感,和特定的诊断方法,为快速诊断和流行病学调查提供技术支持。
    S. suis is an important zoonotic pathogen from sick and recessive carrier pigs that poses a serious threat to animal husbandry production and public health. It usually causes horizontal transmission among pigs. The morbidity and mortality of this disease are very high. Human infection is caused through direct or indirect contact with sick pigs. The two large-scale outbreaks in China were due to the outbreak of S. suis on pig farms, which spread to human infection; thus, detecting S. suis in pig herds is crucial. At present, the commercial S. suis ELISA type 2 kits on the market can only detect single serotypes, high probabilities of interaction reactions, and biosafety risks when using inactivated S. suis as an antigen. Phosphate-3-glyceraldehyde dehydrogenase (GAPDH), muramidase-released protein (MRP), and dihydrolipoamide dehydrogenase (DLDH) are important S. suis type 2, S. suis type 7, and S. suis type 9 protective antigens. This study purified the GMD protein (B-cell-dominant epitopes of GAPDH, MRP, and DLDH antigens) and used a diverse combination of dominant epitopes of the multiple different antigens as coated antigens, improving the sensitivity and safety of the indirect ELISA experiments. An indirect ELISA method (GMD-ELISA) was developed for detecting S. suis antibodies. The antigen-antibody response was optimized using checkerboard titration. The results of testing using ELISA for Salmonella enterica (S. enterica), Escherichia coli (E. coli), Staphylococcus aureus (SA), and Streptococcus pyogenes (S. pyogenes) were all negative, indicating that this method had strong specificity. The results were still positive when the dilution ratio of S. suis-positive serum reached 1:6, 400, thus indicating that the method had high sensitivity. The results of the reproducibility assay for indirect ELISA showed that the intra-assay coefficient of variation and the inter-assay coefficient of variation were less than 10%, indicating that the method had good repeatability. We investigated the seroprevalence of S. suis in 167 serum samples collected in East China, and 33.5% of the samples were positive for antibodies against S. suis, indicating that the prevalence of S. suis is high in pig farms in Eastern China. The novel GMD-ELISA is a convenient, sensitive, and specific diagnostic method that provides technical support for rapid diagnosis and epidemiological investigation.
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  • 文章类型: Journal Article
    简介2019年新型冠状病毒病(COVID-19)是全球关注的主要问题。由于无症状病例的数量未经测试,感染者的实际比例可能高于报告的患病率.因此,通过血清学调查调查产生抗病毒抗体的人的确切比例对于确定人群的免疫状况和相应的指导公共卫生决策至关重要。目的本研究的目的是估计社区中SARS-CoV-2的血清阳性率,并描述发现病例的流行病学特征。方法在2020年7月至10月之间,进行了横断面血清调查,包括从沙特阿拉伯七个地区收集的15,873份血清样本。使用多级方便采样,邀请人们参加由面试官管理的问卷。之后,收集血样并使用SARS-CoV-2病毒IgG/IgM抗体检测试剂盒(ELISA)测定血清阳性率.使用<0.05和95%CI的p值报告显著性。结果样本中SARS-CoV-2的总血清阳性率为17.0%,麦加地区的反应性病例数最高(33.3%)。除糖尿病外,所有合并症与症状之间存在显着关联。此外,年龄,教育,国籍,和地区都是血清结果的重要预测因素。此外,与确诊或疑似病例接触会使血清阳性的风险增加近1.5倍.结论本研究估计沙特阿拉伯SARS-CoV-2的全国血清阳性率为17%。在这项研究的时候,大多数人群没有SARS-CoV-2特异性抗体.这表明人口仍低于群体免疫力的阈值,并强调了大规模疫苗接种计划和遵守推荐的预防预防措施的重要性。
    Introduction  The new coronavirus disease 2019 (COVID-19) is a major global concern. Due to the number of asymptomatic cases that go untested, the actual proportion of those who have been infected is likely to be higher than the reported prevalence. Thus, investigating the exact proportion of those who developed antibodies against the virus through serological surveys is crucial to identify the immune status of the population and direct public health decisions accordingly. Objectives The aim of this study is to estimate the seroprevalence of SARS-CoV-2 in the community and to describe the epidemiological characteristics of the discovered cases. Methods Between July and October 2020, a cross-sectional sero-survey was conducted including a total of 15,873 serum samples collected from seven regions within the kingdom. Using a multistage convenient sampling, people were invited to participate in an interviewer-administrated questionnaire. Afterward, blood samples were collected and seroprevalence was determined using the SARS-CoV-2 virus IgG/IgM antibody detection kits (ELISA). A p-value of <0.05 and 95% CI were used to report the significance. Results The overall seroprevalence of SARS-CoV-2 in the sample was 17.0%, and Makkah region constituted the highest number of reactive cases (33.3%). There was a significant association between all comorbidities and having symptoms except for diabetes. In addition, age, education, nationality, and region were all significant predeterminants of sero-result. Also, contact with a confirmed or suspected case increased the risk of being seropositive by nearly 1.5 times. Conclusion This study estimated the national seroprevalence of SARS-CoV-2 in Saudi Arabia to be 17%. At the time of this study, most of the population did not have the SARS-CoV-2 specific antibodies. This suggests that the population is still below the threshold of herd immunity and emphasizes the importance of mass vaccination programs and abiding by recommended prevention precautions.
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  • 文章类型: Journal Article
    Getah病毒(GETV),在甲病毒属和Togaviridae家族中,已经在世界各地被发现。GETV导致新生仔猪的高发病率和死亡率,造成严重的经济损失。因此,GETV检测的实验工作十分必要。然而,由于各种不可避免的因素的影响,ELISA法用于动物疾病初筛的检测结果准确度较低。因此,我们优化了基于重组E2(rE2)蛋白的酶联免疫吸附测定(ELISA),用于检测猪血清中的GETV抗体。成功表达E2蛋白,并用SDS-PAGE纯化。来自感染猪的血清的Western印迹分析显示与对应于E2糖蛋白的〜46KDa的病毒抗原的强烈反应。通过使用棋盘滴定并比较P/N值,我们发现包被抗原的最佳浓度为24.5μg/mL,血清标本的最佳稀释度为1:100。辣根过氧化物酶(HRP)缀合的山羊抗猪免疫球蛋白(IgG)的最佳工作稀释度为1:5000。最佳涂覆条件为4℃下12小时。血清标本的最佳培养条件,阻塞,与二级抗体反应均在37°C下1小时。我们还调查了在华东地区收集的133份血清标本中GETV的血清阳性率,而37.59%的样本经检测抗GETVIgG抗体呈阳性,表明GETV在中国猪种群中的血清阳性率很高。春季血清阳性率显着降低(4月;24.24%,16/66)比秋季(10月;50.75%,34/67),这表明猪中抗GETV抗体的存在是季节性的。总之,我们改进了rE2ELISA,可在实验和自然感染后检测猪针对GETV的抗体。这在GETV感染的诊断和监测中应该是有用的。
    Getah virus (GETV), in the genus Alphavirus and the family Togaviridae, has been detected throughout the world. GETV causes high morbidity and mortality in newborn piglets, entailing serious economic losses. Therefore, the experimental work on GETV detection is necessary. However, due to the influence of a variety of unavoidable factors, the ELISA test for the primary screening of animal diseases has low accuracy in detection results. Therefore, we optimized a recombinant E2 (rE2) protein-based enzyme-linked immunosorbent assay (ELISA) for the detection of GETV antibodies in pig serum. The E2 protein was successfully expressed and purified with SDS-PAGE. A Western blotting analysis of sera from infected pigs showed strong reaction with a viral antigen of ~46 KDa corresponding to the E2 glycoproteins. By using chessboard titration and comparing the P/N values, we found that the optimal concentration of coated antigen was found to be 24.5 μg/mL, and the optimal dilution of serum specimens was 1:100. The best working dilution of the horseradish peroxidase (HRP)-conjugated goat anti-pig immunoglobulin (IgG) was 1:5000. The optimal coating conditions were 12 h at 4 °C. The optimal incubation conditions for serum specimens, blocking, and reaction with the secondary antibody were all 1 h at 37 °C. We also investigated the seroprevalence of GETV in 133 serum specimens collected in Eastern China, and 37.59% of the samples tested positive for anti-GETV IgG antibodies, indicating that the seroprevalence of GETV is high in pig populations in China. The seroprevalence was significantly lower in spring (April; 24.24%, 16/66) than in autumn (October; 50.75%, 34/67), which suggested that the presence of anti-GETV antibodies in pigs was seasonal. In conclusion, we improved an rE2 ELISA that detected pig antibodies against GETV after experimental and natural infections. This should be useful in the diagnosis and surveillance of GETV infections.
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