Enzyme-Linked Immunosorbent Assay

酶联免疫吸附试验
  • 文章类型: Journal Article
    UNASSIGNED: Most studies on viral infections among livestock handlers have focused on occupational exposure from inadvertent contact with infected animals. Consequently, little emphasis is given to the effect of their lifestyle on the acquisition of other blood-borne viruses.
    UNASSIGNED: To determine the prevalence and assess risk factors for HIV, HBV and HCV infections among livestock handlers in Ibadan, Nigeria.
    UNASSIGNED: Blood samples were collected from 265 livestock handlers between October 2016 to April 2017 in Ibadan. The samples were tested for the presence of antibodies to HIV and HCV; and surface antigen to HBV using ELISA. Structured questionnaire was administered to collect information on risk factors associated with the transmission of these viruses. Data analysis was carried out using Chi-square test and logistic regression to determine the association between risk factors and predictors of infection (p < 0.05).
    UNASSIGNED: Of 265 participants, 11 (4.2%), 29 (10.9%) and 13 (4.9%) individuals tested positive for HIV, HBV and HCV infections respectively. Two (0.8%) of the participants were coinfected with HIV and HBV while 1(0.4%) was coinfected with both HBV and HCV. Individuals who travelled frequently in the course of Livestock trades had a higher rate of HIV infection.
    UNASSIGNED: A high Infection with HIV, HBV and HCV is common among the study participants. There is a need for continued surveillance and awareness creation on preventive measures against these viruses.
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  • 文章类型: Journal Article
    为了确定SCD中血管闭塞事件的数量是否与岩藻糖基转移酶7(FUT7)的血浆浓度有关,它催化选择素配体的合成。
    预期,分析研究。
    三级医疗保健中心的血液学和化学病理学部门。
    13-45岁的稳态HbSS个体,20例患者有3例或3例以上血管闭塞危象,前一年需要入院治疗(有或没有SCD并发症);17例其他HbSS患者有0-1例血管闭塞危象,前一年需要入院治疗,无疾病并发症。
    比较了通过ELISA测得的FUT7的稳态血浆浓度,这些患者在前一年有一次血管闭塞危象需要住院治疗,但没有疾病并发症,以及有或没有并发症的>3次危象。
    每位HbSS患者的FUT7血浆水平和血管闭塞事件的数量。
    前一年血管闭塞危象>3例患者血浆FUT7浓度均值+标准差为8.6+2.7ng/ml,0-1例危象无并发症者为7.3+1.7ng/ml;独立样本t检验,p>0.05,差异不显著。
    岩藻糖基转移酶7的血浆浓度与镰状细胞病中血管闭塞事件的数量无关。
    没有声明。
    UNASSIGNED: To determine if the number of vaso-occlusive events in SCD relates to plasma concentration of fucosyltransferase 7 (FUT7), which catalyses the synthesis of selectin ligands.
    UNASSIGNED: A prospective, analytical study.
    UNASSIGNED: Haematology and Chemical Pathology Departments of tertiary healthcare centres.
    UNASSIGNED: Steady state HbSS individuals aged 13-45 years, 20 had 3 or more vaso-occlusive crises that required hospital admission in the previous year (with or without complications of SCD); 17 other HbSS persons had 0-1 vaso-occlusive crisis that required hospital admission in the previous year and no disease complications.
    UNASSIGNED: Steady-state plasma concentrations of FUT7 measured by ELISA were compared between SCD patients who had one vaso-occlusive crisis requiring hospital treatment in the previous year but no disease complications and those who had >3 crises with or without complications.
    UNASSIGNED: Plasma level of FUT7and the number of vaso-occlusive events in each HbSS patient.
    UNASSIGNED: Mean + standard deviation plasma concentration of FUT7 was 8.6 + 2.7 ng/ml in patients with >3 vasoocclusive crises in the previous year and 7.3 + 1.7 ng/ml in those with 0-1 crisis and no complications; independent sample t-test, p > 0.05, not significantly different.
    UNASSIGNED: Plasma concentration of fucosyltransferase7 is not associated with the number of vaso-occlusive events in sickle cell disease.
    UNASSIGNED: None declared.
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  • 文章类型: Journal Article
    评估性激素结合球蛋白(SHBG)测定作为研究人群中妊娠糖尿病(GDM)的诊断指标的性能。
    分析横断面研究。
    以医院为基础,贝努埃州立大学教学医院(BSUTH),Makurdi,尼日利亚。
    在BSUTH进行产前护理的24至28周胎龄单胎妊娠妇女,Makurdi.
    在诊断性75克口服葡萄糖耐量试验(OGTT)期间,通过ELISA测定血清SHBG水平,以评估连续选择的符合纳入标准的参与者队列中的GDM。
    参与者的血清SHBG水平和GDM的存在。
    血清SHBG与GDM的存在显着负相关(rpb=-0.534,p值<0.001)。其具有0.897的ROC曲线下面积(95%置信区间=0.858-0.935;p值<0.001)。在研究人群中,指示GDM的临界值为452.0nmol/L,诊断优势比为21.4。
    SHBG是研究人群中GDM的有价值的诊断指标。
    没有声明。
    UNASSIGNED: To assess the performance of the Sex Hormone-Binding Globulin (SHBG) assay as a diagnostic indicator of Gestational Diabetes Mellitus (GDM) in the study population.
    UNASSIGNED: Analytical cross-sectional study.
    UNASSIGNED: Hospital-based, Benue State University Teaching Hospital (BSUTH), Makurdi, Nigeria.
    UNASSIGNED: Women with singleton pregnancies at 24 to 28 weeks gestational age attending Antenatal care at BSUTH, Makurdi.
    UNASSIGNED: Serum SHBG levels were assayed by ELISA during a diagnostic 75-gram Oral Glucose Tolerance Test (OGTT) for assessment of GDM in the cohort of consecutively selected participants who met the inclusion criteria.
    UNASSIGNED: Serum levels of SHBG and presence of GDM in the participants.
    UNASSIGNED: Serum SHBG was significantly negatively correlated (rpb = - 0.534, p-value < 0.001) with the presence of GDM. It had an area under the ROC curve of 0.897 (95% Confidence Interval = 0.858-0.935; p-value < 0.001). A cut-off value of 452.0 nmol/L indicative of GDM had a diagnostic odds ratio of 21.4 in the study population.
    UNASSIGNED: SHBG is a valuable diagnostic indicator for GDM in the study population.
    UNASSIGNED: None declared.
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  • 文章类型: Journal Article
    纳米抗体(Nbs)是免疫测定中的强大工具。然而,它们的小尺寸和单价特性对实际应用提出了挑战。多元化成为解决这些限制的重要策略,提高纳米抗体在免疫测定中的利用率。在这里,我们报告了通过将纳米抗体与铁蛋白融合来构建沙门氏菌特异性fenobody(Fb),产生自组装的24价纳米笼结构。与常规纳米抗体相比,非诺抗体表现出35倍的亲合力增加,同时保持良好的热稳定性和特异性。利用这一进步,使用Fb作为捕获抗体设计了三种ELISA模式,以及未修饰的Nb422(FbNb-ELISA),生物素化Nb422(FbBio-ELISA),和噬菌体展示Nb422(FbP-ELISA)作为检测抗体,分别。值得注意的是,FbNb-ELISA显示检测限(LOD)为3.56×104CFU/mL,比FbBio-ELISA低16倍,与FbP-ELISA相似。此外,通过用腔代替TMB显色底物,开发了一种非诺体和纳米体夹心化学发光酶免疫测定法(FbNb-CLISA),导致LOD减少12倍。总的来说,铁蛋白显示技术代表了一种有前途的方法,用于增强基于纳米抗体的夹心ELISA的检测性能,从而扩大了Nbs在食品检测和其他需要多价修饰的领域中的适用性。
    Nanobodies (Nbs) serve as powerful tools in immunoassays. However, their small size and monovalent properties pose challenges for practical application. Multimerization emerges as a significant strategy to address these limitations, enhancing the utilization of nanobodies in immunoassays. Herein, we report the construction of a Salmonella-specific fenobody (Fb) through the fusion of a nanobody to ferritin, resulting in a self-assembled 24-valent nanocage-like structure. The fenobody exhibits a 35-fold increase in avidity compared to the conventional nanobody while retaining good thermostability and specificity. Leveraging this advancement, three ELISA modes were designed using Fb as the capture antibody, along with unmodified Nb422 (FbNb-ELISA), biotinylated Nb422 (FbBio-ELISA), and phage-displayed Nb422 (FbP-ELISA) as the detection antibody, respectively. Notably, the FbNb-ELISA demonstrates a detection limit (LOD) of 3.56 × 104 CFU/mL, which is 16-fold lower than that of FbBio-ELISA and similar to FbP-ELISA. Moreover, a fenobody and nanobody sandwich chemiluminescent enzyme immunoassay (FbNb-CLISA) was developed by replacing the TMB chromogenic substrate with luminal, resulting in a 12-fold reduction in the LOD. Overall, the ferritin-displayed technology represents a promising methodology for enhancing the detection performance of nanobody-based sandwich ELISAs, thereby expanding the applicability of Nbs in food detection and other fields requiring multivalent modification.
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  • 文章类型: Journal Article
    背景:据报道,印度不同州有几例寨卡病毒病的散发性病例和暴发。
    目的:本文探讨了寨卡病毒(ZIKV)在印度中部博帕尔地区持续传播的可能性,这种疾病的最后一次爆发是在2018年。
    方法:我们筛选了一组75例发热患者,这些患者的局部地方性发热原因如登革热检测呈阴性,基孔肯雅,肠热,疟疾,斑疹伤寒和两组无症状健康个体,以献血者(n=75)和产前母亲(n=75)为代表。我们使用实时聚合酶链反应(PCR)测试了发热患者的血液样本中的ZIKVRNA,对于健康的人来说,我们使用酶联免疫吸附试验测定了抗寨卡免疫球蛋白G(IgG)抗体。
    结果:在通过实时PCR测定法测试的75个样品中的任何一个中均未检测到ZIKVRNA。在自愿献血者和产前母亲中,共发现10例(15.38%)和5例(6.66%)抗ZIKVIgG抗体血清阳性,分别。与血清阴性个体26.60(±5.12)岁相比,血清阳性组的年龄更高33.06(±10.83)岁(P=0.037)。
    结论:这项研究,这是印度抗寨卡抗体血清阳性率的首次调查,报告健康人群中抗寨卡抗体的总血清阳性率为10%,暗示一个正在进行的,低水平,ZIKV在当地社区的无声传播。
    BACKGROUND: Several sporadic cases and outbreaks of Zika virus disease have been reported from different states of India.
    OBJECTIVE: This paper explored the possibility of any ongoing transmission of Zika virus (ZIKV) in the Bhopal region of Central India, where the last outbreak of this disease was reported in 2018.
    METHODS: We screened a group of 75 febrile patients who had already tested negative for the locally endemic causes of fever like dengue, chikungunya, enteric fever, malaria, and scrub typhus and two groups of asymptomatic healthy individuals represented by blood donors (n = 75) and antenatal mothers (n = 75). We tested blood samples of febrile patients for ZIKV RNA using real-time polymerase chain reaction (PCR), and for the healthy individuals, we determined anti-zika immunoglobulin G (IgG) antibodies using enzyme-linked immunosorbent assay.
    RESULTS: ZIKV RNA was not detected in any of the 75 samples tested by real-time PCR assay. Among the voluntary blood donors and antenatal mothers, a total of 10 (15.38%) and 5 (6.66%) individuals were found to be seropositive for anti-ZIKV IgG antibodies, respectively. The seropositive group was found to have higher age 33.06 (±10.83) years as compared to seronegative individuals 26.60 (±5.12) years (P = 0.037).
    CONCLUSIONS: This study, which is the first survey of seroprevalence of anti-Zika antibodies from India, reports an overall seropositivity rate of 10% for anti-Zika antibodies among the healthy population, suggesting an ongoing, low level, silent transmission of ZIKV in the local community.
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  • 文章类型: Journal Article
    定量免疫测定,如传统的酶联免疫吸附测定(ELISA),用于确定未知抗原浓度的基质中抗原的浓度。磁性免疫测定,例如LuminexxMAP技术,允许同时检测多种分析物,并提供更高的灵敏度,特异性,样品体积要求低,和高通量能力。这里,我们描述了使用LuminexMAGPIX®系统的定量免疫测定,以确定未知浓度的液体样品中的抗原浓度。详细来说,我们描述了一种新开发的测定方法,用于确定昆虫细胞培养衍生的上清液中果蝇S2产生的马尔堡病毒(MARV)糖蛋白的产量。该测定的潜在应用可以扩展到定量来自感染活MARV的体外和体内模型的液体中的病毒抗原。从而为病毒学研究提供额外的应用。
    Quantitative immunoassays, such as the traditional enzyme-linked immunosorbent assay (ELISA), are used to determine concentrations of an antigen in a matrix of unknown antigen concentration. Magnetic immunoassays, such as the Luminex xMAP technology, allow for the simultaneous detection of multiple analytes and offer heightened sensitivity, specificity, low sample volume requirements, and high-throughput capabilities. Here, we describe a quantitative immunoassay using the Luminex MAGPIX® System to determine the antigen concentration from liquid samples with unknown concentrations. In detail, we describe a newly developed assay for determining production yields of Drosophila S2-produced Marburg virus (MARV) glycoprotein in insect-cell-culture-derived supernatant. The potential applications of this assay could extend to the quantification of viral antigens in fluids derived from both in vitro and in vivo models infected with live MARV, thereby providing additional applications for virological research.
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  • 文章类型: Journal Article
    单克隆抗体在疾病治疗和抗原检测中有着广泛的应用。它们传统上是使用哺乳动物细胞表达系统生产的,不能大规模满足这些蛋白质日益增长的需求。杆状病毒表达载体系统(BEVS)是用于产生生物活性单克隆抗体的有吸引力的替代平台。在这一章中,我们证明了在BEVS中产生HIV-1广泛中和抗体b12。包括转移载体构建在内的过程,重组杆状病毒一代,描述了抗体的产生和检测。
    Monoclonal antibodies have widespread applications in disease treatment and antigen detection. They are traditionally produced using mammalian cell expression system, which is not able to satisfy the increasing demand of these proteins at large scale. Baculovirus expression vector system (BEVS) is an attractive alternative platform for the production of biologically active monoclonal antibodies. In this chapter, we demonstrate the production of an HIV-1 broadly neutralizing antibody b12 in BEVS. The processes including transfer vector construction, recombinant baculovirus generation, and antibody production and detection are described.
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  • 文章类型: Journal Article
    野生动物,与家畜分享病原体,在传染病的流行病学中起着至关重要的作用。从野生动物中取样带来了重大挑战,然而,纳入疾病监测和监测计划至关重要。通常,大规模监测涉及使用酶联免疫吸附测定(ELISA)测试的血清学筛查,通常只对家畜进行验证。这项研究评估了市售ELISA测试对342份野生反刍动物血清样品和100份野猪的诊断特异性。我们评估了三种口蹄疫测试:两种用于小反刍动物,两个是裂谷热,一个是羊痘病毒。使用公式真阴性/(假阳性+真阴性)计算诊断特异性。科恩的卡帕系数测量测试之间的一致性。结果显示所有测试的高特异性和一致性。口蹄疫(FMD)的特异性范围从Prionics的93.89%到IDEXX的100%,IDvet显示99.6%。FMDIDvet和IDEXX之间的一致性最高,为97.1%。裂谷热(RVF)测试,Ingezim和IDvet,达到100%和98.83%的特异性,分别。通过重新测试单个反应器并灭活补体获得最佳特异性。贡献:市售ELISA试剂盒对口蹄疫和类似的跨界动物疾病具有特异性,可用于高度特异性的野生动物测试。
    Wild animals, sharing pathogens with domestic animals, play a crucial role in the epidemiology of infectious diseases. Sampling from wild animals poses significant challenges, yet it is vital for inclusion in disease surveillance and monitoring programmes. Often, mass surveillance involves serological screenings using enzyme-linked immunosorbent assay (ELISA) tests, typically validated only for domestic animals. This study assessed the diagnostic specificity of commercially available ELISA tests on 342 wild ruminant serum samples and 100 from wild boars. We evaluated three tests for foot-and-mouth disease: two for Peste des petits ruminants, two for Rift Valley fever and one for Capripox virus. Diagnostic specificity was calculated using the formula True Negative/(False Positive + True Negative). Cohen\'s kappa coefficient measured agreement between tests. Results showed high specificity and agreement across all tests. Specificity for foot-and-mouth disease (FMD) ranged from 93.89% for Prionics to 100% for IDEXX, with IDvet showing 99.6%. The highest agreement was between FMD IDvet and IDEXX at 97.1%. Rift Valley fever (RVF) tests, Ingezim and IDvet, achieved specificities of 100% and 98.83%, respectively. The optimal specificity was attained by retesting single reactors and inactivating the complement.Contribution: Commercially available ELISA kits are specific for foot-and-mouth disease and similar transboundary animal diseases and can be used for highly specific wild animal testing.
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  • 文章类型: Journal Article
    诊断方法的整合有望在流行和非流行地区推进疟疾传播的监测。血清学检测是鉴别和界定疟疾传播的有价值的工具,作为快速诊断测试(RDT)和厚涂片显微镜的补充方法。这里,我们评估了针对包含PvMSP-1Sal-I菌株的整个氨基酸序列的肽的抗体作为间日疟原虫暴露的可行血清学生物标志物的潜力。
    我们筛选了包含间日疟原虫孢子表面蛋白1(PvMSP-1)Sal-I菌株的完整氨基酸序列的肽作为间日疟原虫暴露的潜在生物标志物。这里,使用SPOT合成技术,然后进行免疫印迹,鉴定了由感染间日疟原虫的个体的抗体特异性识别的免疫显性肽。在免疫印迹测定中,基于它们的较高且特异的反应性来选择两个15聚体肽。随后,使用SPPS(固相肽合成)以可溶性形式合成肽p70和p314,并通过ELISA(IgG,和子类)。
    这项研究揭示了来自巴西亚马逊地区的大多数间日疟原虫感染个体中针对肽p314的IgG抗体的存在。计算机B细胞表位预测进一步支持利用p314作为评估疟疾传播的潜在生物标志物。由于其氨基酸序列是PvMSP-1保守区块的一部分而得到加强。的确,与感染恶性疟原虫的患者和从未接触过疟疾的未感染个体相比,间日疟原虫感染的患者通过IgG1和IgG3对p314的识别明显更高。
    UNASSIGNED: The integration of diagnostic methods holds promise for advancing the surveillance of malaria transmission in both endemic and non-endemic regions. Serological assays emerge as valuable tools to identify and delimit malaria transmission, serving as a complementary method to rapid diagnostic tests (RDT) and thick smear microscopy. Here, we evaluate the potential of antibodies directed against peptides encompassing the entire amino acid sequence of the PvMSP-1 Sal-I strain as viable serological biomarkers for P. vivax exposure.
    UNASSIGNED: We screened peptides encompassing the complete amino acid sequence of the Plasmodium vivax Merozoite Surface Protein 1 (PvMSP-1) Sal-I strain as potential biomarkers for P. vivax exposure. Here, immunodominant peptides specifically recognized by antibodies from individuals infected with P. vivax were identified using the SPOT-synthesis technique followed by immunoblotting. Two 15-mer peptides were selected based on their higher and specific reactivity in immunoblotting assays. Subsequently, peptides p70 and p314 were synthesized in soluble form using SPPS (Solid Phase Peptide Synthesis) and tested by ELISA (IgG, and subclasses).
    UNASSIGNED: This study unveils the presence of IgG antibodies against the peptide p314 in most P. vivax-infected individuals from the Brazilian Amazon region. In silico B-cell epitope prediction further supports the utilization of p314 as a potential biomarker for evaluating malaria transmission, strengthened by its amino acid sequence being part of a conserved block of PvMSP-1. Indeed, compared to patients infected with P. falciparum and uninfected individuals never exposed to malaria, P. vivax-infected patients have a notably higher recognition of p314 by IgG1 and IgG3.
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  • 文章类型: Journal Article
    目的:这项前瞻性队列研究的目的是评估种植体周围表型(PPh)对种植体周围疾病严重程度的影响以及非手术机械治疗(NSMT)的结果,以及钙卫蛋白(CLP)和MMP-8(基质金属蛋白酶-8)水平。
    方法:纳入39例患者的77个植入物。植入物被归类为第1组(植入物周围粘膜炎),第2组(种植体周围炎)。基线(0。月-PrT)临床参数(PD,GI,PI,防喷器,CAL)和影像学记录骨丢失,并收集种植体周围泪液(PICF)样本。采用各种仪器和方法来评估PPh成分(粘膜厚度,上肌组织高度,角化粘膜)和种植体周围附着粘膜(AM)。将NSMT应用于患病的植入部位。通过在治疗后第6个月(PT)取PICF样品再次评估所有临床参数。在从两组获得的PICF样本中,使用ELISA测试评估MMP-8和CLP水平。
    结果:PrT-PD,PrT-GI,第2组的PrT-CAL和PrT-BOP百分比值显著高于第1组。PrT-PD,在薄生物型植入物中,PrTPI评分显著较高。在瘦生物型中,PPh和AM的所有成分均显着降低。两组MMP-8和CLP的时间依赖性变化均有统计学意义(p<0.05)。当评估薄和厚生物型与生化参数之间的关系时,PrT-PT的变化没有显着差异(p>0.05)。
    结论:PPh在影响种植体周围疾病的严重程度中起作用。然而,表型对NSMT结局的影响在两组中相似.
    结论:计划植入手术时应考虑PPh。
    OBJECTIVE: The purpose of this prospective cohort study is to evaluate the effect of peri-implant phenotype (PPh) on the severity of peri-implant diseases and the results of non-surgical mechanical treatment (NSMT), along with calprotectin (CLP) and MMP-8(matrix metalloproteinase-8) levels.
    METHODS: 77 implants from 39 patients were included. The implants were categorized Group-1(peri-implant mucositis), Group-2(peri-implantitis).Baseline (0. Month-PrT) clinical parameters (PD, GI, PI, BOP, CAL) and radiographic bone loss were documented, and peri-implant crevicular fluid (PICF) samples were collected. Various intruments and methodologies were employed to assess PPh components (mucosa thickness, supracrestal tissue height, keratinized mucosa) and peri-implant attached mucosa (AM). NSMT was applied to diseased implant sites. All clinical parameters were reassessed again by taking PICF samples at the 6th month-after treatment (PT). In PICF samples obtained from both groups, MMP-8 and CLP levels were evaluated using the ELISA test.
    RESULTS: PrT-PD,PrT-GI,PrT-CAL and PrT-BOP percentage values in Group-2 were significantly higher than Group-1.PrT-PD,PrTPI scores are significantly higher in thin biotype implants. All components of the PPh and AM were significantly lower in thin biotype. Intra-group time-dependent changes of MMP-8 and CLP were significant in both groups (p < 0.05). When the relationship between thin and thick biotype and biochemical parameters was evaluated, the change in PrT-PT didn\'t show a significant difference (p > 0.05).
    CONCLUSIONS: PPh plays a role in influencing the severity of peri-implant diseases. However, the impact of phenotype on NSMT outcomes was similar in both groups.
    CONCLUSIONS: The PPh should be considered when planning implant surgery.
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