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 HIV 感染的结核病患者中异质耐药结核分枝杆菌 (MTB) 感染的方法。异抗性 MTB 感染包括耐药型和药物敏感型 MTB。感染异质耐药结核病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。艾滋病毒感染者似乎特别容易感染异质耐药性结核病，而结核病仍然是包括博茨瓦纳在内的资源贫乏国家艾滋病毒感染者死亡的主要原因。测序技术（深度测序）的最新发展可能成为快速检测异耐药 MTB 感染的强大工具。然而，尚无研究系统评估深度测序在临床环境中检测结核病患者异质耐药性的作用。提高对异质耐药性的认识将有助于显着降低结核病发病率和死亡率，并促进对结核病的科学认识。我们建议对博茨瓦纳正在进行的一项基于人群的结核病研究中登记的结核病/艾滋病毒合并感染患者进行探索性亚组研究。我们将识别并招募 200 名新诊断的结核病患者接受一线治疗。我们将在入组时和治疗 2 个月后收集痰样本。我们将使用 MTB DNA 深度测序来寻找异质抗性。我们的提案有两个具体目标。目标 1. 我们将确定与 MTB 培养样本测序相比，痰标本直接测序是否可以增加异质耐药 MTB 感染的检测。目标 2. 我们将确定异耐药 MTB 感染状态是否可以预测一线结核病治疗患者对结核病治疗的反应。这项拟议的研究将是第一个系统评估使用深度测序来检测异质耐药 MTB 感染的研究，通过允许快速、早期检测异质耐药感染并相应地告知治疗决策，有可能改善临床实践。

 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.