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: Journal Article
    肾移植后BK病毒(BKV)感染可引起BKV肾病(BKVAN),导致移植物功能障碍和同种异体移植物丢失。BKVAN的治疗是减少免疫抑制负荷,这会增加肾移植排斥的风险。除了BK病毒载量之外,没有生物标志物来监测BKV活性。系统研究了酶联免疫吸附斑点(ELISPOT)测定法作为监测移植后受体抗BKV免疫反应的工具的价值。电子数据库,包括MEDLINE,Scopus,并在Cochrane对照试验中央注册中心进行了ELISPOT评估BKV免疫反应的研究。BKV状态分为“活动BKV感染”和“解决BKV感染”。进行随机效应模型荟萃分析以确定ELISPOT测定的诊断性能,根据ELISPOT阳性和阴性结果将患者分为几组。确定了一百二十七篇文章,其中九篇被包括在内。ELISPOT阴性的患者BKV复制活跃的风险增加(比值比为71.9(95%-CI31.0-167.1)。合并敏感性为0.95(95%-CI0.89-0.98),特异性为0.88(95%-CI0.78-0.94)。与已解决BKV感染的患者相比,活动性BKV感染的患者之间产生IFN-γ的细胞数量的标准化平均差异为-2.09(95%-CI-2.50,-1.68)。ELISPOT分析是BKV风险评估的有用工具,与BKV负荷结合可以支持临床医生指导BKV复制患者的免疫抑制治疗。
    BK virus (BKV) infection after kidney transplantation can cause BKV nephropathy (BKVAN) resulting in graft dysfunction and allograft loss. The treatment for BKVAN is reduction of the immunosuppressive load which increases the risk of kidney transplant rejection. There is no biomarker to monitor BKV activity besides BK viral load. The value of the Enzyme-Linked Immunosorbent Spot (ELISPOT) assay as a tool to monitor the recipient\'s anti-BKV immune response after transplantation was investigated systematically. Electronic databases, including MEDLINE, Scopus, and the Cochrane Central Register of Controlled Trials were searched for studies of ELISPOT evaluating the immune response against BKV. BKV status was categorized as \"active BKV infection\" and as \"resolving BKV infection\". Random-effects model meta-analysis was performed to determine the diagnostic performance of the ELISPOT assay, after stratifying patients into groups based on positive and negative ELISPOT results. One-hundred twenty-seven articles were identified of which nine were included. Patients with negative ELISPOT had an increased risk of having active BKV replication (odds ratio of 71.9 (95%-CI 31.0-167.1). Pooled sensitivity was 0.95 (95%-CI 0.89-0.98) and specificity was 0.88 (95%-CI 0.78-0.94). The standardized mean difference of the number of IFN-γ producing cells between patients with active BKV infection compared with patients who had resolving BKV infection was -2.09 (95%-CI -2.50, -1.68). The ELISPOT assay is a useful tool for BKV risk assessment and in combination with BKV load may support clinicians in guiding immunosuppressive therapy in patients with BKV replication.
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  • 文章类型: Journal Article
    Acute rejection remains an important problem after kidney transplantation. Enzyme-linked immunosorbent spot (ELISPOT) assay has been investigated extensively and has shown promising results as a predictor of allograft rejection. The objective of this study was to systematically review and analyze the predictive value of the donor-specific ELISPOT assay to identify recipients at risk for acute rejection. Electronic databases were searched for studies reporting donor-specific ELISPOT and kidney transplantation outcomes. Odds ratio (OR) for acute rejection was calculated, along with standardized mean difference (SMD) of cytokine producing-cells between recipients with and without acute rejection. Pooled estimates were calculated using random-effect models. The positive ELISPOT cutoff frequencies were extracted from each study. From 665 articles found, 32 studies were included in the meta-analysis. IFN-γ was the most investigated cytokine (30 out of 32 studies). Patients with positive pre-transplantation donor-reactive IFN-γ ELISPOT had an OR of 3.3 for acute rejection (95%-CI 2.1 to 5.1), and OR of 6.8 (95%-CI 2.5 to 18.9) for post-transplantation ELISPOT. Recipients with rejection had significantly higher frequencies of pre- and post-transplantation cytokine producing-cells (SMD 0.47, 95%-CI 0.07 to 0.87 and SMD 3.68, 95%-CI 1.04 to 6.32, respectively). Pre-transplantation ELISPOT had a positive predictive value of 43% and a negative predictive value of 81% for acute rejection. A positive ELISPOT result was associated with a lower estimated glomerular filtration rate (SMD -0.59, 95%-CI -0.83 to -0.34). In conclusion, patients with a high frequency of donor-reactive IFN-γ ELISPOT are at higher risk for acute rejection. The donor-specific IFN-γ ELISPOT assay can serve as an immune-monitoring tool in kidney transplantation.
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  • 文章类型: Comparative Study
    BACKGROUND: The relative diagnostic accuracy of interferon γ release assays (IGRAs; based on ELISA [enzyme-linked immunosorbent assay] or ELISPOT [enzyme-linked immunosorbent spot], ie, the QuantiFERON and T-SPOT.TB tests, respectively) and the tuberculin skin test (TST) for latent tuberculosis (TB) infection in people with end-stage kidney disease is uncertain and national guidelines for their use are inconsistent.
    METHODS: Systematic review.
    METHODS: Evaluated performance of tests for latent TB with clinical risk-factor assessment.
    METHODS: People with end-stage kidney disease (chronic kidney disease stage 5 [eGFR <15] or kidney transplant recipients). No limits on setting.
    METHODS: ELISA- or ELISPOT-based IGRAs, TST, assays to detect antimycobacterial antibodies, and flow cytometry-based tests.
    RESULTS: Odds of test positivity with clinical risk factor for latent TB, expressed as ORs and relative ORs (RORs).
    RESULTS: 47 studies (6,828 participants) were included, but only 30 studies (4,546 participants) contained sufficient data to contribute to meta-analysis. Studies were predominately in the dialysis population (23/30; 3,700 participants) in countries with low to moderate TB prevalence (0.0-50.0 cases/10(5) persons). BCG vaccination rate was variable (2.7%-100.0%). 9 studies compared IGRAs with the TST directly, 17 studies evaluated the TST only, and the other 4 studies evaluated other tests. Compared to a positive TST result, a positive ELISA-based IGRA result was associated more strongly with radiologic evidence of past TB (ROR, 4.29; 95% CI, 1.83-10.3; P = 0.001) and contact with active TB (ROR, 3.36; 95% CI, 1.61-7.01; P = 0.001). Compared to a negative TST result, a negative ELISA-based IGRA result was associated more strongly with BCG vaccination (ROR, 0.30; 95% CI, 0.14-0.63; P = 0.002). There were insufficient data to compare performance of the ELISPOT-based IGRA with the TST or ELISA-based IGRA.
    CONCLUSIONS: 17 of 47 included studies (36.2%) did not contain sufficient data to contribute to meta-analysis.
    CONCLUSIONS: Compared to the TST, the ELISA-based IGRA was associated more strongly with risk factors for latent TB in end-stage kidney disease.
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  • 文章类型: Editorial
    The role of interferon-gamma release assays (IGRAs) for immunologic diagnosis of tuberculosis in children is under debate. We carried out a narrative review on the studies on IGRAs in paediatric populations. A literature search was conducted using multiple keywords and standardized terminology in Medline, EMBASE and Cochrane databases, up to January 27th, 2011. Study quality was assessed using the MOOSE checklist and results of relevant studies were summarized. Sixty-seven paediatric studies (study population ranging from 14 to 5,244 children) were identified. Non-commercial ELISPOT assay (by means of ESAT-6 and CFP-10 antigens) had been carried out in 11 studies. QuantiFERON-TB Gold (QFT-G), QuantiFERON-TB Gold In-tube (QFT-G-IT), and T-SPOT.TB assays had been performed in 10, 44 and 18 studies, respectively. Most studies reported higher specificity of IGRA than tuberculin skin test (TST), but interpretation of the results is complicated by the fact that a gold standard for the diagnosis of latent TB is lacking. The reported sensitivity for active TB ranged from 51-93 percent for QFT-G/QFT-G-IT and 40-100 percent for ELISPOT assays, suggesting that a negative IGRA result may not exclude tuberculosis. Combining TST and IGRA results increased the diagnostic sensitivity. Rates of indeterminate results largely varied (0 to 35 percent). Most of the studies on young (less than 5 years) or immune-compromised children reported a proportion of indeterminate results exceeding 4 percent. Agreement among TST and IGRA, assessed by the k statistics, ranged from -0.03 to 0.87. Higher rates of discordance were reported in BCG-vaccinated than in non-BCG-vaccinated children. Studies on children less than 5 years and immunocompromised children reported conflicting results, as did studies on serial IGRA determinations. Despite the large amount of literature data, the role of IGRA in the pediatric population is still unclear, especially in young children. Combined use of TST/IGRA may increase diagnostic sensitivity but interpretation of discordant results remains a challenging issue.
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