Immunodiagnostic

免疫诊断
  • 文章类型: Journal Article
    球菌病是阿根廷某些地区特有的潜在威胁生命的真菌感染。感染是由球虫引起的。主要通过球虫抗体(Ab)检测诊断。访问快速,高度准确的诊断测试对于确保及时的抗真菌治疗至关重要。与其他Ab检测测定相比,sōnaCoccidioidesAb侧流测定(LFA)执行速度更快,需要更少的实验室基础设施和设备,可能为球孢子菌病流行地区的快速病例筛查提供实质性改进;然而,需要对此测试进行验证。因此,我们的目的是评估杆纳球虫Ab(LFA)的分析性能,并比较与抗球虫Ab检测试验的一致性.共检测了103份人血清标本,包括25例来自球孢子菌病患者的标本和78例无球孢子菌病患者的标本。以88%的灵敏度进行了球藻抗体侧流测定(LFA),特异性和准确性为87%。此外,球虫AbLFA与其他抗球虫Ab检测试验有良好的一致性.我们的发现表明,s_naCoccidioidesAbLFA具有令人满意的性能,可能有助于在流行地区诊断球虫菌病。
    Coccidiomycosis is a potentially life-threatening fungal infection endemic to certain regions of Argentina. The infection is caused by Coccidioides spp. and is primarily diagnosed by Coccidioides antibody (Ab) detection. Access to rapid, highly accurate diagnostic testing is critical to ensure prompt antifungal therapy. The sōna Coccidioides Ab Lateral Flow Assay (LFA) performs faster and requires less laboratory infrastructure and equipment compared with other Ab detection assays, potentially providing a substantial improvement for rapid case screening in coccidioidomycosis-endemic regions; however, validation of this test is needed. Thus, we aimed to evaluate the analytical performance of the sōna Coccidioides Ab (LFA) and compare agreement with anti-Coccidioides Ab detection assays. A total of 103 human sera specimens were tested, including 25 specimens from patients with coccidioidomycosis and 78 from patients without coccidioidomycosis. The sōna Coccidioides Ab Lateral Flow Assay (LFA) was performed with a sensitivity of 88%, and specificity and accuracy of 87%. Furthermore, the Coccidioides Ab LFA had good agreement with other anti-Coccidioides Ab detection assays. Our findings suggest the sōna Coccidioides Ab LFA has satisfactory performance and may be useful for diagnosing coccidioidomycosis in endemic regions.
    导出

    更多引用

    收藏

    翻译标题摘要

    我要上传

       PDF(Pubmed)

  • 文章类型: Observational Study
    需要新的诊断工具来快速评估肺结核(PTB)治疗的疗效。这项研究的目的是在巴拉圭进行的一项观察性和横断面队列研究中评估几种免疫生物标志物。
    在开始治疗前(T0)对32例临床和微生物学证实的PTB患者进行了评估,治疗2个月后(T1)和治疗结束时(T2)。在每个时间点血浆IFN-y水平,17促炎和抗炎细胞因子/趋化因子和补体因子C1q,使用QuantiFERON-TB金加和重组耻垢分枝杆菌肝素结合血凝素(rmsHBHA)作为刺激抗原,在未刺激和Mtb特异性刺激的全血样品中评估C3和C4。还评估了全血细胞计数和肝酶测定,并将其与血浆中的生物标志物水平相关联。
    在未受激等离子体中,C1q(P<0.001),C4(P<0.001),血红蛋白(P<0.001),基线时,PTB患者的淋巴细胞比例(P<0.001)和绝对白细胞计数(P=0.01)显着高于治愈患者。发现C1q和C4水平与痰中结核分枝杆菌负荷有关。最后,组合分析确定了血浆宿主特征,包括检测响应于rmsHBHA的C1q和IL-13水平,作为区分PTB患者与治愈的TB谱的工具,AUC为0.92(敏感性94%,特异性79%)。
    这项观察性研究为整个抗结核治疗过程中的宿主免疫反应提供了新的见解,并强调了宿主C1q和HBHA特异性IL-13反应作为替代血浆生物标志物在监测结核病治疗效果中的作用。
    New diagnostic tools are needed to rapidly assess the efficacy of pulmonary tuberculosis (PTB) treatment. The aim of this study was to evaluate several immune biomarkers in an observational and cross-sectional cohort study conducted in Paraguay.
    Thirty-two patients with clinically and microbiologically confirmed PTB were evaluated before starting treatment (T0), after 2 months of treatment (T1) and at the end of treatment (T2). At each timepoint plasma levels of IFN-y, 17 pro- and anti-inflammatory cytokines/chemokines and complement factors C1q, C3 and C4 were assessed in unstimulated and Mtb-specific stimulated whole blood samples using QuantiFERON-TB gold plus and recombinant Mycobacterium smegmatis heparin binding hemagglutinin (rmsHBHA) as stimulation antigen. Complete blood counts and liver enzyme assays were also evaluated and correlated with biomarker levels in plasma.
    In unstimulated plasma, C1q (P<0.001), C4 (P<0.001), hemoglobin (P<0.001), lymphocyte proportion (P<0.001) and absolute white blood cell count (P=0.01) were significantly higher in PTB patients at baseline than in cured patients. C1q and C4 levels were found to be related to Mycobacterium tuberculosis load in sputum. Finally, a combinatorial analysis identified a plasma host signature comprising the detection of C1q and IL-13 levels in response to rmsHBHA as a tool differentiating PTB patients from cured TB profiles, with an AUC of 0.92 (sensitivity 94% and specificity 79%).
    This observational study provides new insights on host immune responses throughout anti-TB treatment and emphasizes the role of host C1q and HBHA-specific IL-13 response as surrogate plasma biomarkers for monitoring TB treatment efficacy.
    导出

    更多引用

    收藏

    翻译标题摘要

    我要上传

       PDF(Pubmed)

  • 文章类型: Journal Article
    背景:胰腺导管腺癌(PDAC)是一种破坏性疾病,需要精确诊断才能有效治疗。然而,糖抗原19-9(CA19-9)的诊断价值有限。因此,本研究旨在鉴定用于PDAC诊断的新型肿瘤相关自身抗体(TAAbs).
    方法:三阶段策略,包括发现,test,并实施了验证。使用HuProt™人蛋白质组微阵列v3.1筛选49个样品中的潜在TAAbs。随后,通过PDAC中的酶联免疫吸附试验(ELISA)评估了477个样品中潜在的TAAb水平,良性胰腺疾病(BPD),和正常控制(NC),其次是诊断模型的构建。
    结果:在发现阶段,蛋白质微阵列鉴定出167个候选TAAbs。基于生物信息学分析,选择15种肿瘤相关抗原(TAA)用于使用ELISA进一步验证。10个TAAbs在PDAC患者中表现出差异表达(P<0.05)。曲线下面积(AUC)范围为0.61至0.76。包括三种TAAbs(抗HEXB,抗TXLNA,然后开发了抗SLAMF6),显示AUC为0.81(58.0%灵敏度,86.0%特异性)和0.78(55.71%灵敏度,87.14%特异性)用于区分PDAC和NC。此外,该模型的AUC为0.80(灵敏度为58.0%,86.25%特异性)和0.83(55.71%灵敏度,100%特异性)用于在测试和验证阶段区分PDAC和BPD,分别。值得注意的是,免疫诊断模型与CA19-9的组合导致PDAC阳性率增加至92.91%.
    结论:免疫诊断模型可能为PDAC诊断提供一种新的血清学检测方法,为开发有效的诊断生物标志物提供有价值的见解。
    BACKGROUND: Pancreatic ductal adenocarcinoma (PDAC) is a devastating disease that requires precise diagnosis for effective treatment. However, the diagnostic value of carbohydrate antigen 19 - 9 (CA19-9) is limited. Therefore, this study aims to identify novel tumor-associated autoantibodies (TAAbs) for PDAC diagnosis.
    METHODS: A three-phase strategy comprising discovery, test, and validation was implemented. HuProt™ Human Proteome Microarray v3.1 was used to screen potential TAAbs in 49 samples. Subsequently, the levels of potential TAAbs were evaluated in 477 samples via enzyme-linked immunosorbent assay (ELISA) in PDAC, benign pancreatic diseases (BPD), and normal control (NC), followed by the construction of a diagnostic model.
    RESULTS: In the discovery phase, protein microarrays identified 167 candidate TAAbs. Based on bioinformatics analysis, fifteen tumor-associated antigens (TAAs) were selected for further validation using ELISA. Ten TAAbs exhibited differentially expressed in PDAC patients in the test phase (P < 0.05), with an area under the curve (AUC) ranging from 0.61 to 0.76. An immunodiagnostic model including three TAAbs (anti-HEXB, anti-TXLNA, anti-SLAMF6) was then developed, demonstrating AUCs of 0.81 (58.0% sensitivity, 86.0% specificity) and 0.78 (55.71% sensitivity, 87.14% specificity) for distinguishing PDAC from NC. Additionally, the model yielded AUCs of 0.80 (58.0% sensitivity, 86.25% specificity) and 0.83 (55.71% sensitivity, 100% specificity) for distinguishing PDAC from BPD in the test and validation phases, respectively. Notably, the combination of the immunodiagnostic model with CA19-9 resulted in an increased positive rate of PDAC to 92.91%.
    CONCLUSIONS: The immunodiagnostic model may offer a novel serological detection method for PDAC diagnosis, providing valuable insights into the development of effective diagnostic biomarkers.
    导出

    更多引用

    收藏

    翻译标题摘要

    我要上传

       PDF(Pubmed)

  • 文章类型: Journal Article
    抗体代表了一种相对成熟的检测手段,在癌症的临床诊断和治疗中作为治疗药物载体,其中单克隆抗体(mAb)目前占据主导地位。然而,由于单克隆抗体的诸多局限性,小分子单域抗体的出现和发展是不可避免的,比如它们的大尺寸,复杂结构,对极端温度敏感,和肿瘤微环境。因此,自从1995年在软骨鱼类中首次发现以来,IgNAR已成为替代单克隆抗体的替代治疗策略,因此,这种新型免疫球蛋白在临床诊断和肿瘤治疗方面受到了广泛关注。IgNAR的可变新抗原受体(VNAR)因其体积小,为新型抗肿瘤药物的开发提供了优势,高稳定性,亲和力高,以及其他结构和功能特征。在这方面,需要更好地了解IgNAR/VNAR在临床和抗肿瘤治疗中的独特特征和治疗潜力.本文综述了其独特的生化条件和分子结构在临床诊断和新型抗肿瘤药物方面的优势。同时,现有结合药物的主要优点,基于单结构域抗体,在这里介绍,从而为今后临床诊断和抗肿瘤治疗的发展提供新的思路和方法。
    Antibodies represent a relatively mature detection means and serve as therapeutic drug carriers in the clinical diagnosis and treatment of cancer-among which monoclonal antibodies (mAbs) currently occupy a dominant position. However, the emergence and development of small-molecule monodomain antibodies are inevitable due to the many limitations of mAbs, such as their large size, complex structure, and sensitivity to extreme temperature, and tumor microenvironments. Thus, since first discovered in Chondroid fish in 1995, IgNAR has become an alternative therapeutic strategy through which to replace monoclonal antibodies, thus entailing that this novel type of immunoglobulin has received wide attention with respect to clinical diagnoses and tumor therapies. The variable new antigen receptor (VNAR) of IgNAR provides an advantage for the development of new antitumor drugs due to its small size, high stability, high affinity, as well as other structural and functional characteristics. In that respect, a better understanding of the unique characteristics and therapeutic potential of IgNAR/VNAR in clinical and anti-tumor treatment is needed. This article reviews the advantages of its unique biochemical conditions and molecular structure for clinical diagnoses and novel anti-tumor drugs. At the same time, the main advantages of the existing conjugated drugs, which are based on single-domain antibodies, are introduced here, thereby providing new ideas and methods for the development of clinical diagnoses and anti-tumor therapies in the future.
    导出

    更多引用

    收藏

    翻译标题摘要

    我要上传

       PDF(Pubmed)

  • 文章类型: Journal Article
    囊性包虫病(CE),由细粒棘球蚴引起的,是一种主要的人畜共患疾病,会导致大量的人类发病率和死亡率。这种世界性疾病很难诊断,请客,和控制。到目前为止,含有抗原B或抗原5的包虫囊液的粗提物已被用作其免疫诊断的主要抗原来源。主要问题是它与感染其他蠕虫的人的血清反应。目前没有标准,具体,或用于疾病诊断的敏感测试,并且没有人类疫苗的报道。
    考虑到需要有效的免疫和/或免疫诊断,六种细粒大肠杆菌抗原,抗原5,抗原B,热休克蛋白如Hsp-8和Hsp-90,磷酸烯醇丙酮酸羧激酶,和tetraspanin-1被选择。
    使用各种硅片工具,通过靶向抗原5,抗原B,预测T细胞和B细胞表位(混杂肽)。热休克蛋白如Hsp-8和Hsp-90,磷酸烯醇丙酮酸羧激酶,和tetraspanin-1.
    有12种混杂肽与人类白细胞抗原(HLA)I类重叠,二级,和构象B细胞表位。这种免疫显性肽可用作亚单位疫苗。此外,还发现了六种特异于细粒大肠杆菌的肽,这可能是诊断CE的重要标志,可能防止误诊和管理不善。
    这些表位可能是颗粒大肠杆菌中最重要的疫苗靶标,因为它们具有最混杂的肽和B细胞表位,以及对不同等位基因的最高亲和力,由对接分数确定。然而,使用体外和体内模型进行额外的研究。
    UNASSIGNED: Cystic echinococcosis (CE), caused by Echinococcus granulosus, is a major zoonotic disease that causes significant human morbidity and mortality. This cosmopolitan disease is difficult to diagnose, treat, and control. So far, crude extracts of hydatid cyst fluid containing antigen B or antigen 5 have been used as the primary antigenic source for its immunodiagnosis. The main issue is that it reacts with sera from people infected with other helminths. There is currently no standard, specific, or sensitive test for disease diagnosis, and no human vaccine has been reported.
    UNASSIGNED: Considering the need for efficient immunization and/or immunodiagnosis, six E. granulosus antigens, antigen 5, antigen B, heat shock proteins such as Hsp-8 and Hsp-90, phosphoenolpyruvate carboxykinase, and tetraspanin-1, were chosen.
    UNASSIGNED: Using various in silico tools, T cell and B cell epitopes (promiscuous peptides) were predicted by targeting antigen 5, antigen B, heat shock proteins such as Hsp-8 and Hsp-90, phosphoenolpyruvate carboxykinase, and tetraspanin-1.
    UNASSIGNED: There are twelve promiscuous peptides with overlapping human leukocyte antigen (HLA) class-I, class-II, and conformational B cell epitopes. Such immunodominant peptides could be useful as subunit vaccines. Furthermore, six peptides specific for E. granulosus were also discovered, which may prove to be important markers in the diagnosis of CE, potentially preventing misdiagnosis and mismanagement.
    UNASSIGNED: These epitopes may be the most important vaccine targets in E. granulosus because they have the most promiscuous peptides and B cell epitopes, as well as the highest affinity for different alleles, as determined by docking scores. However, additional research using in vitro and in vivo models is undertaken.
    导出

    更多引用

    收藏

    翻译标题摘要

    我要上传

       PDF(Pubmed)

  • 文章类型: Journal Article
    背景:血吸虫病是由血吸虫属吸虫引起的一种被忽视的热带寄生虫病,影响全球约2.4亿人。该疾病的诊断可以通过寄生虫学进行,分子,和/或免疫学方法,然而,开发新的诊断方法对于指导政策决策仍然至关重要,监测疾病趋势并评估干预措施的有效性。
    目的:在这个意义上,目前的工作总结了关于酶联免疫吸附试验中应用的抗原的系统综述的结果,在过去的十年中获得了专利并出版了。
    方法:文献检索策略使用医学主题标题(网格)术语定义为描述符。“曼氏血吸虫”与描述符“免疫测定”一起使用,“酶联免疫吸附测定”,\"elisa\",和“抗原”,使用\"和\"连接器。专利搜索是使用关键字完成的,包括诊断和血吸虫病或血吸虫病或血吸虫。多个数据库用于专利检索,如知识产权国家研究所;欧洲专利局;美国专利商标局;专利范围,谷歌专利。
    结果:检索到41篇文章,其中只有五个符合资格标准。从数据库中获得了17项专利,和最相关的发明的简要描述在这里给出。
    结论:血吸虫病被认为是世界上最重要的蠕虫病。因此,寻找和发展基于血清学的诊断方法,以降低该疾病的发病率和死亡率是重要的。
    Schistosomiasis is a neglected tropical parasitic disease caused by trematode worms of the genus schistosoma, which affects approximately 240 million people worldwide. the diagnosis of the disease can be performed by parasitological, molecular, and/or immunological methods, however, the development of new diagnostic methods still essential to guide policy decisions, monitor disease trends and assess the effectiveness of interventions.
    in this sense, the current work summarizes the findings of a systematic review regarding antigens applied in the enzyme-linked immunosorbent assay test, which were patented and published over the last ten years.
    the literature search strategy used medical subject heading (mesh) terms to define as descriptors. \"schistosoma mansoni\" was used in arrangement with the descriptors \"immunoassay\", \"enzyme-linked immunosorbent assay\", \"elisa\", and \"antigens\", using the \"and\" connector. the patent search was done using keywords, including diagnosis and schistosoma or schistosomiasis or schistosome. several databases were employed for the patent search, such as intellectual property national institute; european patent office; the united states patent and trademark office; patent scope, and google patents.
    forty-one articles were retrieved, of which only five met the eligibility criteria. seventeen patents were taken from the databases, and a brief description of the most relevant inventions is given here.
    schistosomiasis is considered the most important helminthic disease in worldwide. therefore, it is important to of searching for and develops diagnostic methods based on serology to reduce morbidity and mortality caused by the disease.
    导出

    更多引用

    收藏

    翻译标题摘要

    我要上传

    求助全文

  • 文章类型: Journal Article
    检测KPC型碳青霉烯酶对于指导适当的抗生素治疗以及实施抗生素管理和感染控制措施是必要的。目前,很少有测试能够区分碳青霉烯酶类型,限制实验室报告是否存在。这项工作的目的是产生抗体并开发ELISA测试以检测KPC-2及其D179突变体。使用兔和小鼠多克隆抗体设计ELISA-KPC测试。测试了四种不同的方案以选择具有最高灵敏度和特异性的细菌接种物。标准化程序是使用109个先前表征的临床分离株进行的,显示100%的灵敏度和89%的特异性。ELISA-KPC检测到所有产生碳青霉烯酶的分离株,包括显示ESBL表型的KPC变体,例如KPC-33和-66。
    The detection of KPC-type carbapenemases is necessary for guiding appropriate antibiotic therapy and the implementation of antimicrobial stewardship and infection control measures. Currently, few tests are capable of differentiating carbapenemase types, restricting the lab reports to their presence or not. The aim of this work was to raise antibodies and develop an ELISA test to detect KPC-2 and its D179 mutants. The ELISA-KPC test was designed using rabbit and mouse polyclonal antibodies. Four different protocols were tested to select the bacterial inoculum with the highest sensitivity and specificity rates. The standardisation procedure was performed using 109 previously characterised clinical isolates, showing 100% of sensitivity and 89% of specificity. The ELISA-KPC detected all isolates producing carbapenemases, including KPC variants displaying the ESBL phenotype such as KPC-33 and -66.
    导出

    更多引用

    收藏

    翻译标题摘要

    我要上传

       PDF(Pubmed)

  • 文章类型: Journal Article
    内脏利什曼病(VL)是由利什曼原虫属的细胞内原生动物引起的感染的致命表现。在新世界国家,VL被归类为人畜共患疾病,家犬是其主要水库。无症状的狗与有症状的狗一样有能力将利什曼原虫传播给载体,然而,目前的诊断测试是有限的,对这一重要人群的敏感性较低。准确测试的发展是早期诊断的基础,治疗,和控制犬利什曼病。在这项研究中,我们研究了重组蛋白(dynamin-1-likeprotein,Dyn-1)来自L.婴儿,作为利什曼病血清诊断有症状和无症状犬的潜在靶抗原。通过使用来自症状(n=25)的血清的ELISA测定来评估蛋白质的抗原性能,使用ELISA的无症状(n=34)和未感染的狗(n=36)。此外,实验感染克氏锥虫(n=49)并自然感染巴贝虫的狗的血清。测试(n=8)以评估可能的交叉反应性。利什曼原虫的粗可溶性抗原(CSA)用作抗原对照,而K39和K26用作参考抗原,因为它们已经广泛用于商业测试。与抗原K39(88%)相比,基于rDyn-1的检测显示出最高的灵敏度(97%),K26(86%)和粗提物(95%)。rDyn-1蛋白(94%)也获得了测试中最高的特异性,与其他抗原K39(81%)相比,K26(87%),和粗提物(77%)。这项研究表明,rDyn-1ELISA测定法能够识别100%的无症状犬,确立其作为犬利什曼病诊断目标的潜力。
    Visceral leishmaniasis (VL) is a fatal manifestation of an infection caused by intracellular protozoa of the Leishmania genus. In New World countries, VL is classified as a zoonotic disease with domestic dogs acting as its main reservoir. Asymptomatic dogs are as competent to transmit Leishmania to the vectors as symptomatic dogs, however current diagnostic tests are limited and present low sensitivity for this important group. The development of accurate tests is fundamental to the early diagnosis, treatment, and control of canine leishmaniasis. In this study, we investigated the use of a recombinant protein (dynamin-1-like protein, Dyn-1) from L. infantum, as a potential target antigen for leishmaniasis serodiagnosis in both symptomatic and asymptomatic dogs. The antigenic performance of the protein was evaluated by means of ELISA assays using sera from symptomatic (n = 25), asymptomatic (n = 34) and non-infected dogs (n = 36) using ELISA. In addition, sera from dogs experimentally infected with Trypanosoma cruzi (n = 49) and naturally infected with Babesia sp. (n = 8) were tested to evaluate possible cross-reactivity. A crude soluble antigen (CSA) of Leishmania was used as an antigen control and K39 and K26 were used as reference antigens because they are already widely used in commercial tests. rDyn-1-based assay showed the highest sensitivity (97%) compared to the antigens K39 (88%), K26 (86%) and crude extract (95%). The highest specificity among the tests was also obtained with the protein rDyn-1 (94%), compared with the other antigens K39 (81%), K26 (87%), and crude extract (77%). This study showed that the rDyn-1 ELISA assay was able to identify 100% of asymptomatic dogs, establishing its potential as a target for the diagnosis of canine leishmaniasis.
    导出

    更多引用

    收藏

    翻译标题摘要

    我要上传

    求助全文

  • 文章类型: Journal Article
    人类弓形虫病是一种被忽视的蠕虫病,它是由人畜共患的round虫物种犬弓形虫和弓形虫引起的。人类弓形虫病的诊断是基于临床,寄生虫学,和流行病学标准,以及检测抗弓形虫抗体的血清学测试。尽管如此,由于缺乏疾病的病理症状和体征,血清学是支持结论性诊断的关键证据。TES-ELISA是用于诊断的最广泛使用的血清学测试。然而,TES抗原与其他蠕虫抗原产生的抗体的交叉反应是其在寄生虫患病率高的国家应用的主要缺点。犬T.canis重组抗原已被描述为用于诊断的天然TES的替代物。然而,抗原蛋白的选择是一个复杂的过程,需要验证。在本文中,我们开发了一种基于eGFP载体的系统来表达和纯化T.canis抗原蛋白的重组多肽块。通过免疫印迹检测到强烈的交叉反应多肽,并避免最终产生嵌合原型蛋白。此外,产生了包含完整测试蛋白质的对照嵌合蛋白质。纯化的嵌合抗原用阴性和阳性对照个体的310个血清样品在ELISA和免疫印迹测定中进行测试。我们的结果表明,嵌合rCHITC0和rCHITC1抗原(敏感性为62%58%,在IB-rCHITC0,ELISA-rCHITC0,ELISA-rCHITC1和IB-rCHITC1中,OLMS分别为38%和16%)在特异性方面表现更好(为91%,89%,对于OLMS,ELISA-rCHITC1,IB-rCHITC1,ELISA-rCHITC0和IB-rCHITC0分别为87%和76%)比T.canisTES-ELISA(特异性为61%),在受感染个体的血清样本中给出更高的信号,以及区分其他寄生虫感染的假阳性病例的可能性。我们的数据表明T.canis嵌合蛋白,代表II期研究的候选抗原。
    Human toxocariasis is one of the neglected helminthiases and it is caused by the zoonotic roundworm species Toxocara canis and Toxocara cati. Diagnosis of human toxocariasis is based on the combination of clinical, parasitological, and epidemiological criteria, as well as serology tests that detect anti-Toxocara antibodies. Notwithstanding, due to the absence of pathognomonic symptoms and signs of the disease, serology is the key evidence to support a conclusive diagnosis. TES-ELISA is the most widely used serological test for diagnosis. However, cross-reaction of TES antigens with antibodies produced to other helminth antigens is a major drawback for its application in countries with high parasitic prevalence. T. canis recombinant antigens have been described as an alternative to native TES for diagnosis. Nevertheless, the selection of antigenic proteins is a complex process that requires validation. In this paper, we developed an eGFP carrier-based system to express and purify blocks of recombinant polypeptides of T. canis antigenic proteins. Intense cross-reaction polypeptides were detected by Immunoblot and avoided to finally produce a chimeric prototype protein. Additionally, a control chimeric protein that harbors the complete tested proteins was produced. Purified chimeric antigens were tested in ELISA and Immunoblot assays with 310 sera samples of negative and positive control individuals. Our results showed that chimeric rCHITC0 and rCHITC1 antigens (with sensitivities of 62% 58%, 38% and 16% in IB-rCHITC0, ELISA-rCHITC0, ELISA-rCHITC1 and IB-rCHITC1 respectively for OLMS) can perform better in terms of specificity (being 91%, 89%, 87% and 76% for ELISA-rCHITC1, IB-rCHITC1, ELISA-rCHITC0 and IB-rCHITC0 respectively for OLMS) than T. canis TES-ELISA (with 61% specificity), giving a higher signal with serum samples of infected individuals as well the possibility to discriminate false positive cases with other parasitic infections. Our data suggest that T. canis chimeric proteins, represent candidate antigens for phase II studies.
    导出

    更多引用

    收藏

    翻译标题摘要

    我要上传

       PDF(Pubmed)

  • 文章类型: Journal Article
    疟疾是全球健康和残疾的主要原因之一,即使在努力根除它之后。迄今为止,尚无高效疫苗可用于其控制。疫苗效力低的主要原因是人群中潜在疫苗候选抗原基因的广泛多态性和HLA多态性。这个问题可以通过开发使用混杂肽结合HLA等位基因数量的疫苗来解决。本研究通过靶向间日疟原虫的PPPK-DHPS和DHFR-TS蛋白预测T和B细胞表位(混杂肽),使用不同的硅片工具。所选择的肽根据其免疫原性被表征为混杂肽。抗原性和疏水性。此外,来确认它们的免疫原性,这些肽被用于分子建模和对接分析。为了确定与不同HLA-I类的必要亲和力,和HLAII类等位基因,仅选择5种用于DHFR-TS的肽和3种用于PPPK-DHPS的肽作为混杂肽。D1肽与HLA等位基因具有最大的结合能,根据HLA-肽复合物建模和结合相互作用分析。这些发现可能导致开发具有改善的安全性和有效性的基于表位的疫苗接种。这些表位可能是间日疟原虫的主要疫苗靶标,因为它们具有较高数量的混杂肽。此外,如通过对接评分所分析的,B细胞表位对不同等位基因具有最大亲和力。然而,需要在体外和体内进行进一步的研究.
    Malaria is one of the major causes of health and disability globally, even after tremendous efforts to eradicate it. Till date no highly effective vaccine is available for its control. The primary reason for the low efficacy of vaccines is extensive polymorphism in potential vaccine candidate antigen genes and HLA polymorphisms in the human population. This problem can be resolved by developing a vaccine using promiscuous peptides to combine the number of HLA alleles. This study predicted T and B cell epitopes (promiscuous peptides) by targeting PPPK-DHPS and DHFR-TS proteins of Plasmodium vivax, using different in silico tools. Selected peptides were characterized as promiscuous peptides on the basis of their immunogenicity, antigenicity and hydrophobicity. Furthermore, to confirm their immunogenicity, these peptides were utilized for molecular modelling and docking analysis. For determining the requisite affinity with distinct HLA Class-I, and HLA Class-II alleles, only five peptides for DHFR-TS and 3 peptides for PPPK-DHPS were chosen as promiscuous peptides. The D1 peptide has the maximum binding energy with HLA alleles, according to HLA-peptide complex modelling and binding interaction analyses. These findings could lead to the development of epitope-based vaccinations with improved safety and efficacy. These epitopes could be major vaccine targets in P. vivax as they possess a higher number of promiscuous peptides. Also, the B cell epitopes possess maximum affinity towards different alleles as analyzed by docking scores. However, further investigation is warranted in vitro and in vivo.
    导出

    更多引用

    收藏

    翻译标题摘要

    我要上传

    求助全文

公众号