Tuberculosis Diagnostics: Towards More Precise Testing in Children

结核病诊断：对儿童进行更精确的检测

• 批准号: 7470622
• 负责人: JOSEPH Alphonso BELLANTI
• 金额: $ 7.61万
• 依托单位: GEORGETOWN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-08-01 至 2009-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7470622
• 关键词: 5 year old; AIDS/HIV problem; Acid Fast Bacillae Staining Method; Adolescent; Adult; Age; Age-Years; Agreement; Antigens; Australia; Bacillus (bacterium); Binding; Biological Assay; Blood Tests; Calmette-Guerin Bacillus; Cells; Cellular Immunity; Child; Childhood; Clinical; Communicable Diseases; Communities; Contracts; Developed Countries; Developing Countries; Diagnosis; Diagnostic; Diagnostic Procedure; Disadvantaged; Disease; Dose; Effectiveness of Interventions; False Positive Reactions; Gender; Genus Mycobacterium; Goals; Gold; Health Personnel; Homelessness; Hypersensitivity skin testing; Immigrant; Immune; In Vitro; Incidence; Individual; Infant; Infection; Interferon Type II; Jail; Laboratories; Lobe; Lymphocyte; Malaria; Measurement; Measures; Mediating; Methodology; Methods; Mycobacterium tuberculosis; Negative Skin Test; Older Population; Paper; Patients; Peptides; Persons; Pilot Projects; Population; Predisposition; Prevalence; Production; Proteins; Range; Rate; Reading; Reporting; Research; Resources; Severity of illness; Shelter facility; Sputum; Standards of Weights and Measures; Symptoms; Techniques; Testing; Tuberculin; Tuberculosis; United States Food and Drug Administration; Variant; Whole Blood; World Health Organization; age group; base; cell mediated immune response; clinical Diagnosis; cross reactivity; cytokine; disorder prevention; early childhood; improved; in vitro Assay; named group; response; tool

DESCRIPTION (provided by applicant): This is a revised R03 application to develop an improved in vitro assay of cell mediated immunity to measure cytokine production for the diagnosis of tuberculosis in children. The century-old tuberculin skin test remains the standard way of diagnosis of latent tuberculosis (TB) infection, and is often used as an ancillary test in the diagnosis of clinical TB. The skin test requires two separate encounters with a health-care provider, and there is a sizable proportion of persons, particularly in disadvantaged, immigrant populations, who find it difficult to return to have their skin tests read. In addition, the test requires careful application of the test dose, and careful reading to be reliable, and is often done poorly. Clearly, better tests are needed. A whole blood test based upon release of interferon-gamma from sensitized lymphocytes stimulated with PPD antigen (the QuantiFERON-TB test, from Cellestis Ltd. South Melbourne, Australia) had been previously approved by the FDA. Recently a newer version (the QuantiFERON-TB Gold test) has also been approved, but neither test for the pediatric population. The use of the recently available, highly specific pools of ESAT-6 and CFP-10 peptides (included in the QuantiFERON-TB Gold test) which are absent from all BCG strains and from most non-tuberculosis mycobacteria (with the exception of M. kansasii, M. szulgai, M. marinum, M. leprae, M. bovis and M. africanum) should aid diagnosis of tuberculosis and eliminate the problem of cross- reactivity in individuals infected with most non-tuberculous mycobacteria or previously immunized with BCG. In the present studies we shall be using the QuantiFERON-TB Gold Test which contains as individual components the newly available pools of overlapping peptides of ESAT-6 and CFP-10. We shall also employ cocktails of the two plus TB7.7 (p4), as well as RD1- selected peptides of ESAT-6 and CFP-10 provided by Dr. D. Goletti. A set of pilot studies based upon the use of this improved test will be performed in a clinically well pediatric, mostly immigrant population to evaluate its correlation with conventional tuberculin skin testing (TST), and, on a short-term basis, to determine its ability to predict clinical TB. A particular emphasis will be placed on the younger infant-child (1 to 5 year-old) population where both the severity of disease and the need is great. Moreover, since in patients with suspected clinical TB, there may be "false-negative" skin tests due to suppressor cytokines and other mechanisms related to maturational immaturity that may obscure true cell-mediated immunity to MTB, studies will be also included to determine optimal dose- response relationships (particularly important in children) which may permit overcoming this "false- negativity" in the laboratory. Thus, these studies will explore techniques for differentiating false negative and false positive reactions, and hopefully permit more reliable In vitro testing for the determination of true cell- mediated immunity to MTB and possible differentiating latent from active TB disease.

描述（由申请人提供）：这是修订后的 R03 申请，旨在开发改进的细胞介导免疫体外测定法，以测量细胞因子的产生，以诊断儿童结核病。具有百年历史的结核菌素皮试仍然是诊断潜伏性结核感染的标准方法，并且经常被用作临床结核病诊断的辅助试验。皮肤测试需要与医疗保健提供者进行两次单独的接触，并且有相当一部分人，特别是弱势群体和移民群体，发现很难返回进行皮肤测试读取。此外，该测试需要仔细应用测试剂量，并仔细读数才能可靠，但往往做得很差。显然，需要更好的测试。基于 PPD 抗原刺激的致敏淋巴细胞释放干扰素-γ 的全血检测（QuantiFERON-TB 检测，来自澳大利亚南墨尔本的 Cellestis Ltd.）此前已获得 FDA 批准。最近，更新版本（QuantiFERON-TB Gold 测试）也已获得批准，但这两种测试均不适用于儿科人群。使用最近可用的高度特异性的 ESAT-6 和 CFP-10 肽库（包括在 QuantiFERON-TB Gold 测试中），所有 BCG 菌株和大多数非结核分枝杆菌（分枝杆菌除外）均不存在这些肽库。 kansasii、M. szulgai、M. marinum、M. leprae、M. bovis 和 M. africanum）应有助于结核病的诊断并消除以下问题：感染大多数非结核分枝杆菌或之前接受过卡介苗免疫的个体存在交叉反应。在本研究中，我们将使用 QuantiFERON-TB Gold Test，其中包含新可用的 ESAT-6 和 CFP-10 重叠肽池作为单独的成分。我们还将使用两者的混合物加上 TB7.7 (p4)，以及由 D. Goletti 博士提供的 ESAT-6 和 CFP-10 的 RD1 选择肽。一系列基于使用这种改进测试的试点研究将在临床良好的儿科（主要是移民人群）中进行，以评估其与传统结核菌素皮肤测试（TST）的相关性，并在短期内确定其预测临床结核病的能力。将特别重视年龄较小的婴儿（1 至 5 岁）人群，因为他们的疾病严重程度和需求都很大。此外，由于在疑似临床结核病患者中，由于抑制性细胞因子和其他与成熟不成熟相关的机制可能会出现“假阴性”皮试，这可能会掩盖真正的细胞介导的针对结核病的免疫，因此还将包括研究以确定最佳的剂量反应关系（对儿童尤其重要）可能允许克服实验室中的这种“假阴性”。因此，这些研究将探索区分假阴性和假阳性反应的技术，并希望能够进行更可靠的体外测试，以确定真正的细胞介导的MTB免疫，并可能区分潜伏性结核病和活动性结核病。

项目成果

Rapid diagnosis of Mycobacterium tuberculosis infection in children using interferon-gamma release assays (IGRAs).

使用干扰素-γ释放测定（IGRA）快速诊断儿童结核分枝杆菌感染。

• DOI: 10.2500/aap.2012.33.3574
• 发表时间: 2012
• 期刊: Allergy and asthma proceedings
• 影响因子: 2.8
• 作者: Riazi,Shahla;Zeligs,Barbara;Yeager,Henry;Peters,StephenM;Benavides,GermanA;DiMita,Onorina;Bellanti,JosephA
• 通讯作者: Bellanti,JosephA