Utility of Deep Sequencing for Detecting Heteroresistant Mycobacterium tuberculosis Infections among HIV-infected Persons

 描述(由申请人提供)：拟议的研究将有助于建立快速检测艾滋病毒感染结核病患者中耐异基因结核分枝杆菌(MTB)感染的方法。异质性耐药结核杆菌感染既涉及耐药类型的结核杆菌，也涉及药物敏感类型的结核杆菌。对结核分枝杆菌具有异质性耐药感染的人更有可能发展为完全耐药结核病，这可能导致治疗结果不佳，增加发病率和死亡率。艾滋病毒携带者感染异源耐药结核病的风险似乎特别高，在包括博茨瓦纳在内的资源贫乏国家，结核病仍然是艾滋病毒感染者死亡的主要原因。测序技术(深度测序)的最新进展可能是快速检测耐异药结核杆菌感染的有力工具。然而，还没有研究系统地评估在临床环境中检测结核病患者异质性耐药的深度测序。提高对异质性耐药的理解将有助于显著改善结核病发病率和死亡率，并促进对结核病的科学理解。我们建议对博茨瓦纳正在进行的基于人群的结核病研究中登记的结核病/艾滋病毒混合感染患者进行一项探索性的亚研究。我们将识别并招募200名新诊断的结核病患者进行一线治疗。我们将在登记时和治疗2个月后收集痰样本。我们将使用MTB DNA的深度测序来寻找异质性抗性。我们的建议有两个具体目标。目的1.我们将确定与结核杆菌培养样本的直接测序相比，从痰标本中直接测序是否能增加对异源耐药结核杆菌感染的检测。目的2.我们将确定在接受一线结核病治疗的患者中，异源耐药结核分枝杆菌感染状态是否可以预测对结核病治疗的反应。这项拟议的研究将首次系统地评估深度测序技术检测耐异耐药结核杆菌感染的使用，通过允许快速、早期地检测耐异耐药感染并相应地通知治疗决定，潜在地改进临床实践。

 DESCRIPTION (provided by applicant): The proposed study will help to establish methods for rapid detection of heteroresistant Mycobacterium tuberculosis (MTB) infections among HIV-infected TB patients. Heteroresistant MTB infections involve both drug-resistant and drug-susceptible types of MTB. A person with heteroresistant MTB infection is more likely to develop fully drug-resistant TB, which can cause poor treatment outcomes, increasing morbidity and mortality. People living with HIV appear to be especially at high risk for acquiring heteroresistan TB infections, and TB remains the leading cause of death among HIV-infected persons in resource poor countries, including Botswana. Recent developments in sequencing technology (deep sequencing) could be a powerful tool for rapidly detecting heteroresistant MTB infections. However, no study has systematically evaluated deep sequencing for detecting heteroresistance among TB patients in a clinical setting. Improved understanding of heteroresistance will contribute to significant improvements in TB morbidity and mortality and advance scientific understanding of TB. We propose to conduct an exploratory substudy of TB/HIV co-infected patients enrolled in an ongoing population-based TB research study in Botswana. We will identify and enroll 200 newly diagnosed TB patients on first-line therapy. We will collect sputum samples at enrollment and after 2 months of treatment. We will use deep sequencing of MTB DNA to look for heteroresistance. There are two specific aims to our proposal. Aim 1. We will determine whether direct sequencing from sputum specimen increases the detection of heteroresistant MTB infections compared to sequencing done on MTB culture samples. Aim 2. We will determine whether heteroresistant MTB infection status is predictive of response to TB treatment among patients on first-line TB therapy. This proposed research will be the first to systematically evaluate the use of deep sequencing to detect heteroresistant MTB infections, potentially improving clinical practice by allowing for rapid, early detection of heteroresistant infections and informing treatment decisions accordingly.