Pre-clinical Development of an Advanced Genexpert Test for Drug Resistant Mtb

先进的 Genexpert 耐药 Mtb 检测的临床前开发

• 批准号: 8829144
• 负责人: David Alland
• 金额: $ 128.42万
• 依托单位: RBHS-NEW JERSEY MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-04-01 至 2019-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8829144
• 关键词: Accounting; Advanced Development; Algorithms; Amikacin; Aminoglycoside Antibiotics; Aminoglycosides; Antitubercular Agents; Automation; Biological Assay; Chemistry; Clinical; Clinical Trials; Color; Complement; Computer software; Country; Decision Making; Detection; Development; Disease; Drug resistance; Drug resistance in tuberculosis; Dyes; Extreme drug resistant tuberculosis; Fluoroquinolones; Future; Genetic; Grant; Hand; Health; Incidence; Infection Control; Institution; Kanamycin; Laboratories; Methods; Modification; Molecular; Moxifloxacin; Multi-Drug Resistance; Mutation; Mycobacterium tuberculosis; Patients; Performance; Pharmaceutical Preparations; Phase; Predisposition; Preparation; Procedures; Quality Control; Reagent; Reporting; Resistance; Rifampicin resistance; Rifampin; Risk; Robotics; Sampling; Sentinel; Sputum; Temperature; Testing; Therapeutic; Time; Tracer; Tuberculosis; United States National Institutes of Health; Validation; Work; amplification detection; assay development; base; clinically actionable; cost effective; expectation; fluoroquinolone resistance; improved; inhibitor/antagonist; innovation; internal control; isoniazid; killings; manufacturing process; melting; mortality; novel; pathogen; pre-clinical; preclinical study; prevent; reagent testing; research and development; research clinical testing; resistance mutation; tuberculosis drugs

DESCRIPTION (provided by applicant): The need for rapid and sensitive methods to detect drug-resistant tuberculosis (TB) has never been greater. The incidence of multi-drug-resistant (MDR) and extensively-drug resistant (XDR) TB is increasing at an alarming rate. Even in the absence of MDR and XDR TB, rapid Fluoroquinolone (FQ) resistance testing will be increasingly needed for patients who are treated with the Moxifloxacin-based regimes that are expected to partially replace Isoniazid (INH)-based treatment in the future. Our group developed the Xpert MTB/RIF (Xpert) assay, the first ever self-contained, near-patient assay that can perform TB detection and also detect resistance to the drug Rifampicin (RIF). This test is transforming TB detection worldwide by making TB detection simple and rapid. However in regards to drug resistance detection, the Xpert assay is limited to identifying patients who potentially have MDR by using RIF-resistance as a surrogate. The Xpert assay cannot tell whether a patient with RIF resistance remains INH-susceptible, and thus could still be treated with an INH-containing regime. Nor can it identify which RIF-resistant patients can be treated with a FQ and/or aminoglycoside as opposed to those who have XDR TB. Thus, patients are at risk for being either over treated or undertreated, with potentially serious consequences. Furthermore, patients with undiagnosed XDR can remain infectious for prolonged periods, further spreading their disease. Here, we propose to combine highly innovative 10-color real-time PCR detection (that makes use of novel large Stokes-shift dyes), innovative three phase PCR fluidics, sloppy molecular beacon probes, the existing GeneXpert platform and basic Xpert assay cartridge to create a new assay that identifies resistance to INH, the FQs, amikacin and kanamycin, important types of resistance not detected by the Xpert assay. This new "Xtend-XDR" assay will complement the current Xpert assay, making it possible to select appropriate treatment (and infection control) approaches in patients identified with TB. The Xtend-XDR assay will use the same testing procedures and testing platform as the Xpert, and largely be restricted to patients who already test TB positive (and RIF resistant) in a primary Xpert test. These features will make it highly cost effective. We have developed proof of principal for all essential assay components. In the current proposal, we will finalize assay development, and create manufacturing processes, quality control, and assay software. Finally, we will perform the analytic and pre-clinical studies needed to prepare for clinical trials. These activities are exactly the type requested in the current partnership RFA. Our work will be performed in the following specific aims: 1. Finalize all assay parameters and internal control components in wet format. 2. Adapt assay for robotic assembly and modify assembly line for cartridge needs. 3. Develop analytic software and a simple interface for clinical operators. 4. Perform analytic and pre-clinical studies in preparation for clinical trials.

描述（由申请人提供）：需要快速和敏感的方法检测耐药性结核病（TB）从未如此大。多药耐药（MDR）和广泛抗药（XDR）结核病的发生率正在以惊人的速度增加。即使在没有MDR和XDR TB的情况下，对于接受莫西法沙星治疗的患者来说，越来越多地需要快速氟喹诺酮（FQ）耐药性测试，这些患者将来有望部分替代基于异念珠菌（INH）的治疗。 我们的小组开发了XPERT MTB/RIF（XPERT）测定法，这是第一个独立的，近乎病人的测定法，可以执行TB检测并检测到对Rifampicin（RIF）的耐药性。该测试通过使结核病检测简单而迅速地改变了全世界的结核病检测。但是，在耐药性检测方面，XPERT分析仅限于鉴定通过使用RIF抗性作为替代物具有MDR的患者。 XPERT测定法无法判断出具有RIF耐药性的患者是否仍然可感染，因此仍然可以通过含INH的制度进行治疗。与患有XDR TB的患者相反，它也无法确定可以用FQ和/或氨基糖苷治疗哪些耐能力性患者。因此，患者面临过度治疗或不足治疗的风险，并可能带来严重的后果。此外，未诊断XDR的患者长期可以保持感染力，从而进一步传播其疾病。 在这里，我们建议将高度创新的10色实时PCR检测（利用新颖的大型斯托克斯搬运染料），创新性的三期PCR流体，草率的分子信标探测，现有的GenExpert平台和基本XPERT ASSAY Cartridge创建了一种对电阻的新测定，以识别Inh，fqs，amikikikin，Amikikin，amikikin，Kan Amikikin，Kan，Kan， XPERT分析。这种新的“ XTEND-XDR”测定将补充当前的XPERT分析，使得在用TB鉴定的患者中选择适当的治疗方法（和感染控制）方法。 XTEND-XDR分析将使用与XPERT相同的测试程序和测试平台，并且在很大程度上仅限于在初级XPERT测试中已经测试TB阳性（和RIF）的患者。这些功能将使其具有很高的成本效益。 我们已经为所有基本测定部分开发了本金证明。在当前的建议中，我们将最终确定测定开发，并创建制造过程，质量控制和测定软件。最后，我们将进行为临床试验做准备所需的分析和临床前研究。这些活动正是当前合伙RFA中要求的类型。我们的工作将以以下特定目的进行：1。以湿法最终确定所有测定参数和内部控制组件。 2。适应机器人组件和修改装配线的测定法。 3。为临床操作员开发分析软件和简单的接口。 4。进行分析和临床前研究，为临床试验做准备。