就近检测:结合噬菌体和 LAMP 检测结核病、牛结核病和约翰氏病

Christopher G. Shield, Alexandra E. M. Bartlett, Pranabashis Haldar, Benjamin M. C. Swift
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引用次数: 0

摘要

分枝杆菌疾病每年对人类和兽医领域造成数百万人的影响。这些疾病的诊断费时费力,通常只有在疾病晚期才会敏感。因此,人们需要在感染早期阶段就能有效、快速、简便的新诊断方法。本研究详细介绍了对用于检测结核杆菌、牛结核杆菌和副结核杆菌亚种的 LAMP 检测方法的优化,并将其与噬菌体介导的裂解相结合,以满足新型诊断方法--噬菌体-LAMP 的需求。优化后的噬菌体-LAMP 检测方法的检测限低于每毫升 10 个分枝杆菌,且检测方法之间无交叉反应。噬菌体-LAMP 方法随后在少量疑似肺结核病人和疑似约翰氏病牛群的临床血液样本中进行了测试。噬菌体-LAMP 检测法可检测出这些样本中存活的结核分枝杆菌和副结核分枝杆菌。
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Detecting Closer to Care: Combining Phage and LAMP to Detect Tuberculosis, Bovine TB and Johne’s Disease
Mycobacterial diseases impact millions in the human and veterinary fields each year. Their diagnosis is long and laborious, often only sensitive in the late stages of disease. This has created an unmet need for new diagnostics that are effective in the earlier stages of infection and are quick and easy to perform. This study details the optimization of LAMP assays for the detection of M. tuberculosis, M. bovis and M. avium subsp. paratuberculosis combined with phage-mediated lysis to meet the needs of a novel diagnostic—termed phage-LAMP. The optimized phage-LAMP assay had a limit of detection of less than 10 mycobacteria per ml and no cross-reaction was seen between assays. The phage-LAMP method was then tested on a small number of clinical blood samples from suspected TB patients and herds suspected of Johne’s disease. The phage-LAMP assay could detect viable Mycobacterium tuberculosis and M. avium subsp. paratuberculosis in these samples.
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