MAS-PCR Rapid Screening Test for Multidrug-Resistant Tuberculosis (MDR-TB)

描述（由申请人提供）：结核分枝杆菌对用于治疗结核感染的药物的耐药性已成为一个重大问题，并且耐多药结核病（MDR-TB）的存在在全世界范围内不断增加。耐多药结核病被定义为对利福平（RIF）和异烟肼（INH）这两种用于治疗结核病的一线药物均具有耐药性。目前用于检测耐多药结核病的传统方法需要在培养物中培养结核菌，然后针对用于治疗的药物进行测试。这是很费力的，需要几周到几个月的时间才能知道结果，这会延迟开始使用更昂贵的有效二线药物的治疗。与耐多药结核病相关的基因突变已被确定，并且已经开发出几种快速聚合酶链反应（PCR）测试来筛查耐多药结核病。这些基于 PCR 的检测可提供更快速的结果，但价格昂贵，并且需要专门的实验室设备，而结核病高负担国家的大多数实验室无法轻易负担得起这些设备，而这些国家大多是资源有限的发展中国家。拟议研究的主要目的是测试一种简单且廉价的多重等位基因特异性聚合酶链反应（MAS-PCR）技术，该技术可用于在高负担资源有限的国家快速筛查耐多药结核病。 MAS-PCR 检测已在泰国曼谷玛希隆大学医学院微生物学系耐药结核病 (DRTB) 实验室的单一实验室环境中开发和测试。拟议的研究由两部分组成，旨在确认该测试可在多个实验室中使用，并且可以在这些实验室中提供准确且一致的结果。对于该研究的第一部分，已知对 RIF 和 INH 敏感或耐药的结核病模拟样本将被送往泰国和中国的五个实验室。实验室不知道样品是否敏感或耐药。每个实验室的测试结果将与样本的已知状态以及另一种更昂贵的基于 PCR 的测试 Genotype MTBDR Plus 的结果进行比较。在研究的第二部分中，中国和泰国的实验室将接收其医院确诊结核病患者的痰样本，并通过 MAS-PCR 检测对他们进行筛查，以确定他们是否可能是耐多药结核病病例。对当地样本进行的这些测试的结果将通过在 Siriraj 医院 DRTB 实验室使用 Genotype MTBDR Plus 对这些样本的 DNA 样本进行重新测试来确认。如果MAS-PCR筛查试验在泰国和中国的实验室成功实施，它将成为在其他高负担资源有限国家进一步试验和实施耐多药结核病常规筛查的候选方案。

DESCRIPTION (provided by applicant): Resistance of Mycobacterium tuberculosis to drugs used for treatment of TB infection has become a significant problem, and the existence of multidrug-resistant tuberculosis (MDR-TB) is increasing throughout the world. MDR-TB is defined as resistance to both rifampicin (RIF) and isoniazid (INH), two of the first-line drugs used for treatment of TB. The conventional method used today to detect MDR-TB requires growing the TB organism in culture and then testing it against the drugs used for treatment. This is laborious and requires weeks to months before the result is known, which delays starting treatment with more expensive second-line drugs that will be effective. Genetic mutations associated with MDR-TB have been identified, and several rapid polymerase chain reaction (PCR) tests have been developed to screen for MDR-TB. These PCR-based tests provide more rapid results, but they are expensive and require specialized lab equipment that cannot be easily afforded by most labs in high-burden TB countries, which are mostly resource-limited developing countries. The primary objective of the proposed research is to test a simple and inexpensive multiplex allele-specific polymerase chain reaction (MAS-PCR) technique that can be used for rapid screening of MDR-TB in high burden resource-limited countries. The MAS-PCR test has already been developed and tested in a single laboratory environment at the Drug-Resistant Tuberculosis (DRTB) Laboratory, Department of Microbiology, Faculty of Medicine, Siriraj Hospital, Mahidol University, and Bangkok, Thailand. The proposed study consists of two parts designed to confirm that this test is feasible for use in multiple labs and can provide accurate and consistent results across these labs. For the first part of the study, simulated samples of TB with known susceptibility or resistance to RIF and INH will be sent to each of five labs in Thailand and China. The labs will not know whether the samples are susceptible or resistant. Test results from each lab will be compared to the known status of the sample, as well as to the result from another more expensive PCR-based test, the Genotype MTBDR Plus. In the second part of the study, the labs in China and Thailand will receive sputum samples from current patients at their hospitals with confirmed TB and will screen them with the MAS-PCR test to determine whether they might be MDR-TB cases. The results from these tests on local specimens will be confirmed by retesting on the DNA samples from these specimens with Genotype MTBDR Plus at the DRTB Laboratory at Siriraj Hospital. If the MAS-PCR screening test is successfully implemented in the labs in Thailand and China, it will be a candidate for further testing in other high-burden resource limited countries and implementation for routine screening for MDR-TB.