Enzyme-Linked Immunospot Assay

酶联免疫斑点测定
  • 文章类型: Review
    肺结核(TB)在中国很常见,但过去很少有结核病伴凝血障碍和全血细胞减少的报道。在这份报告中,一名70岁的女性因食欲不振而入院,深色尿液,恶心,呕吐,疲劳,双侧下肢水肿;胸部CT提示双肺弥漫性感染性病变,凝血功能障碍,和完全的全血细胞减少症,最初被认为是由严重感染引起的。然而,患者的症状没有通过有效的经验性抗生素治疗得到改善,重复的胸部CT显示肺部病变比以前恶化得更多,凝血障碍和全血细胞减少没有改善。最后,结核病患者使用支气管镜肺泡灌洗技术对结核分枝杆菌(MTB)的酶联免疫斑点试验(ELISPOT)和宏基因组测序(mNGS)检测呈阳性.所以ati-TB是用HRftELfx(异烟肼,0.3gqd;利福喷丁,0.45gbiw;乙胺丁醇,0.75gqd;和左氧氟沙星,0.5gqd)方案。最终,患者的临床症状明显改善,肺部病变被吸收,凝血功能和血细胞计数恢复正常,取得了满意的治疗效果。
    Pulmonary Tuberculosis (TB) is common in China, but tuberculosis with coagulation disorders and pancytopenia have rarely been reported in the past. In this report presented, a 70-year-old female was admitted to the hospital with poor appetite, dark urine, nausea, vomiting, fatigue, and bilateral lower limb edema; chest CT suggested diffuse infectious lesions in both lungs, coagulation dysfunction, and complete pancytopenia, which was initially considered to be caused by severe infection. However, the patient\'s symptoms did not improve by potent empiric antibiotics treatment, and a repeat chest CT showed that the lung lesions deteriorated more than before, and coagulation disorders and pancytopenia did not improve. Finally, the TB patient tested positive for enzyme-linked immunospot assay (ELISPOT) and metagenomic sequencing (mNGS) of Mycobacterium tuberculosis (MTB) using bronchoscopic alveolar lavage. So ati-TB was initiated with HRftELfx (isoniazid, 0.3 g qd; rifapentine, 0.45 g biw; ethambutol, 0.75 g qd; and levofloxacin, 0.5 g qd) regimen. Eventually, the patient\'s clinical symptoms improved significantly, the pulmonary lesions were absorbed, and the coagulation function and blood cell count returned to normal, which achieved a satisfactory treatment effect.
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  • 文章类型: Review
    背景:结核病是一种空气传播的传染病,在胸部影像学上有多种形态学改变。结核病特异性酶联免疫斑点测定(T-SPOT。TB)广泛用于结核病的诊断。临床上,肺结核合并T-SPOT。TB阴性和间质改变非常罕见。
    方法:T-SPOT。TB,致病性测试,胸部CT扫描,下一代测序(NGS)。
    结果:实验室检查显示T-SPOT阴性。TB和痰抗酸染色,胸部CT显示两肺间质纤维化和多个高密度阴影,痰NGS显示结核分枝杆菌感染。
    结论:T-SPOT阴性。TB和间质性肺改变不排除结核分枝杆菌感染。NGS在感染性疾病病原体检测中具有较高的特异性,尤其是在复杂的,混合传染病。
    BACKGROUND: Tuberculosis is an airborne infectious disease with multiple morphologic changes on chest imaging. Tuberculosis-specific enzyme-linked immunospot assay (T-SPOT.TB) is widely used in the diagnosis of tuberculosis. Clinically, pulmonary tuberculosis with T-SPOT.TB negative and interstitial changes is very rare.
    METHODS: T-SPOT.TB, pathogenetic testing, chest CT scan, next-generation sequencing (NGS).
    RESULTS: Laboratory tests showed negative T-SPOT.TB and sputum antacid staining, chest CT showed interstitial fibrosis and multiple high-density shadows in both lungs, and sputum NGS showed Mycobacterium tuberculosis infection.
    CONCLUSIONS: Negative T-SPOT.TB and interstitial lung changes do not exclude Mycobacterium tuberculosis infection. NGS has a high specificity in the detection of pathogens in infectious diseases, especially in complex, mixed infectious diseases.
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  • 文章类型: Case Reports
    An 88-year-old woman was referred to our hospital for autoimmune hepatitis in 2016. She was treated with prednisolone. In 2018, she was rehospitalized owing to hepatitis relapse. Steroid pulse therapy was performed. She exhibited good recovery of hepatitis, but was transferred to a convalescent ward in a general hospital because of decreased activity of daily life. After a month later, she had high fever and cough. She was diagnosed as having tuberculosis because of positive Mycobacterium tuberculosis polymerase chain reaction. At our first medical examination in 2016, we performed enzyme-linked immunospot and the result was undeterminable. There is an increase in the opportunities to use immunosuppressant and biologic agents for elderly patients. Our case report should contribute to future medical care for elderly patients who are at risk of latent tuberculosis infection.
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  • 文章类型: Case Reports
    BACKGROUND: Acute hepatitis E virus (HEV) infections are usually self-limiting in immunocompetent patients. HEV persistence has been described only in immunosuppressed patients such as solid-organ transplant recipients, patients with hematological diseases, or patients with human immunodeficiency virus (HIV) infection.
    UNASSIGNED: A 61-year-old patient was admitted in hospital for jaundice and asthenia.
    UNASSIGNED: The patient had underlying cirrhosis and developed a chronic HEV infection.
    METHODS: Ribavirin therapy was initiated.
    RESULTS: Ribavirin therapy for 12 months allowed the clearance of the virus and HEV viral load remained undetectable thereafter. This patient had taken no immunosuppressive drugs, was not suffering from any autoimmune disease and was not infected with HIV. We studied the patient\'s anti-HEV immune response months after the viral clearance. His peripheral blood mononuclear cells (PBMC) were stimulated in vitro by HEV peptides. The patient had a mild T lymphopenia, but polyclonal stimulation of PBMC showed a robust T cell response. The response of his anti-HEV specific interferon-γ producing T cells was low.
    CONCLUSIONS: Other studies are now needed to identify the population with a chronic evolution of HEV infection despite no apparent immunodepression.
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  • 文章类型: Case Reports
    Host hepatitis C virus (HCV)-specific T cell responses and the ability of the virus to escape this response are important correlates of infection outcome. Understanding this host-viral interplay has been difficult given the often asymptomatic nature of acute HCV infection. We studied a recent transmission case to determine whether adapted viral strains can be transmitted and influence the recipient\'s anti-HCV T cell response. The diversity of viral populations was examined using next-generation sequencing, and HCV-specific T cell interferon (IFN)-γ responses were assessed using a peptide panel representing the autologous viruses. HCV-specific T cell responses in the source were directed against peptides that did not match the dominant autologous virus but rather low-frequency variants, implying existing viral adaptation in the source strain. Most HCV T cell epitopes that elicited an IFN-γ response in the source did not in the recipient, despite the pair sharing human leukocyte antigen alleles that govern antigen presentation and similar autologous viruses. Intrahost HCV variation in the recipient fell within predicted T cell epitopes, suggesting alternative targets of the immune response. These data suggest that transmission of adapted viral species can direct the host\'s HCV-specific immune response profile during acute infection.
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  • 文章类型: Case Reports
    Ewing sarcoma is a highly resistant disease with a <10% chance of survival at 5 years after failure of frontline chemotherapy. This is a case report of an Ewing sarcoma patient with metastatic disease recurrence <2 years after standard chemotherapy/radiation who achieved a durable and sustained complete response after 2 series of treatments with Vigil (GMCSF/bi-shRNA furin DNA autologous tumor immunotherapy) serially manufactured from first and second recurrences with ELISPOT assay correlation. Results support justification of further testing of Vigil with ELISPOT assay as a biomarker to assess level of immune response and correlation with disease control.
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  • 文章类型: Case Reports
    Tuberculosis (TB) remains a major health problem in the world. The clinical forms of TB in children are variable, pulmonary involvement occurs in two thirds of cases. In the remaining third, clinical forms incluye node, meningeal and osteoarticular involvement.
    METHODS: 7 year old boy with a history of an osteolytic lesion of the right ischial branch. Three months later he presented with spondylodiscitis at L2-L3, associated with a large abscess in the right iliac psoas muscle. Pott\'s disease was suspected, and tuberculin test and T-SPOT®.TB test were performed, with a positive result. Antimicrobial treatment was initiated with isoniazid, rifampicin, pyrazinamide and ethambutol. After 30 days, Mycobacterium tuberculosis was isolated from psoas abscess. We discuss methods of TB diagnosis, with special emphasis on immunological methods: tuberculin test and interferon-gamma release assays. Methods of immunological TB diagnosis are an important contribution to the diagnosis of this disease, allowing early initiation of treatment.
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  • 文章类型: Clinical Trial, Phase II
    The first phase IIb safety and efficacy trial of a new tuberculosis vaccine since that for BCG was completed in October 2012. BCG-vaccinated South African infants were randomized to receive modified vaccinia virus Ankara, expressing the Mycobacterium tuberculosis antigen 85A (MVA85A), or placebo. MVA85A did not significantly boost the protective effect of BCG. Cryopreserved samples provide a unique opportunity for investigating the correlates of the risk of tuberculosis disease in this population. Due to the limited amount of sample available from each infant, preliminary work was necessary to determine which assays and conditions give the most useful information. Peripheral blood mononuclear cells (PBMC) were stimulated with antigen 85A (Ag85A) and purified protein derivative from M. tuberculosis in an ex vivo gamma interferon (IFN-γ) enzyme-linked immunosorbent spot assay (ELISpot) and a Ki67 proliferation assay. The effects of a 2-h or overnight rest of thawed PBMC on ELISpot responses and cell populations were determined. Both the ELISpot and Ki67 assays detected differences between the MVA85A and placebo groups, and the results correlated well. The cell numbers and ELISpot responses decreased significantly after an overnight rest, and surface flow cytometry showed a significant loss of CD4(+) and CD8(+) T cells. Of the infants tested, 50% had a positive ELISpot response to a single pool of flu, Epstein-Barr virus (EBV), and cytomegalovirus (CMV) (FEC) peptides. This pilot work has been essential in determining the assays and conditions to be used in the correlate study. Moving forward, PBMC will be rested for 2 h before assay setup. The ELISpot assay, performed in duplicate, will be selected over the Ki67 assay, and further work is needed to evaluate the effect of high FEC responses on vaccine-induced immunity and susceptibility to tuberculosis disease.
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  • 文章类型: Journal Article
    一名24岁有脊柱关节病病史的男性出现高烧,颈淋巴结肿大和泛发性斑丘疹。他在入院前3周用泼尼松龙治疗慢性葡萄膜炎,然后改用柳氮磺吡啶。实验室发现显示明显的白细胞增多,常见的非典型淋巴细胞。停用柳氮磺吡啶,并探讨单核细胞增多症的病因。入院期间,他出现了无结石性胆囊炎和低血压。全身性皮质类固醇给药后,所有症状迅速改善。传染性单核细胞增多症的研究结果为阴性,并使用酶联免疫斑点测定法确认了柳氮磺胺吡啶诱导的超敏反应综合征的诊断。
    A 24-year-old male with a history of spondyloarthropathy presented with high fever, cervical lymphadenopathy and generalized maculopapular rash. He was treated with prednisolone for chronic uveitis before being switched to sulfasalazine 3 weeks prior to admission. Laboratory findings revealed marked leukocytosis with frequent atypical lymphocytes. Sulfasalazine was discontinued and the etiology of mononucleosis syndrome explored. During admission, he developed acalculous cholecystitis and hypotension. All symptoms quickly improved following administration of systemic corticosteroids. The investigation for infectious mononucleosis yielded negative results and a diagnosis of sulfasalazine-induced hypersensitivity syndrome was confirmed using enzyme-linked immunospot assays.
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  • DOI:
    文章类型: English Abstract
    OBJECTIVE: To highlight the clinical features and diagnosis of chronic disseminated tuberculosis, with emphasizing the usefulness of several recently available diagnostic technologies in this setting.
    METHODS: We presented a case of chronic disseminated tuberculosis diagnosed with the combined application of interferon-gamma release assay T-SPOT. TB, 18F-fluorodeoxyglucose (FDG)-positron emission tomography (PET), and gene chip assay.
    RESULTS: A 53-year-old gentleman who had chronic cough for 7 years and fever for 2 weeks was referred to our hospital for further evaluation. 18F-FDG-PET/CT scan showed increased FDG uptake in multiple lesions involving bilateral lungs, supraclavicular, mediastinal and intro-abdominal lymph nodes and bones, mimicking metastatic malignancy. T-SPOT. TB assay revealed significant responses [ early secreting antigen target 6 (ESAT-6): 3 908 spot forming cells (SFCs)/10(6) peripheral blood mononuclear cells (PBMCs), culture filtrate protein (CFP-10): 3 400 SFCs/10(6) PBMCs]. Subsequent biopsy of supraclavicular lymph node, lung, and ilium revealed granulomas, while culture of the obtained tissue yeilded mycobacteria. Gene chip testing identified M. tuberculosis sensitive to isoniazid and rifampin. After 10 weeks of treatment for tuberculosis, the patient\'s condition was improved and a second T-SPOT. TB assay showed significantly reduced responses (ESAT-6: 1528 SFCs/10(6) PBMCs; CFP-10: 1460 SFCs/10(6) PBMCs).
    CONCLUSIONS: Timely diagnosis of chronic disseminated tuberculosis requires high index of suspicion. T-SPOT. TB assay, PET/CT, and gene chip assay may provide valuable information that facilitates further diagnostic procedures and treatment decision.
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