Division of Bioresources, International Institute for Zoonosis Control, Hokkaido University, Sapporo, Hokkaido, Japan.
Department of Pathobiology, Faculty of Veterinary Medicine, Lilongwe University of Agriculture and Natural Resources, Lilongwe, Malawi.
J Vet Diagn Invest. 2023 May;35(3):307-310. doi: 10.1177/10406387231164596. Epub 2023 Apr 8.
The lack of quick, accurate, and low-cost detection methods has hindered the active control strategies for bovine tuberculosis (bTB) in resource-limited countries with a high burden of disease. We developed a dry loop-mediated isothermal amplification (LAMP) assay for rapid and specific detection of , the principal causative agent of bTB, and evaluated the efficacy of the assay using suspected bTB samples collected during routine meat inspection at major regional abattoirs in Malawi. Template genomic DNA was extracted directly from the granulomatous bTB-like lesion (crude extracted DNA), as well as growth from the incubated mycobacterial growth indicator tubes (MGIT). Field results were visualized by the naked eye within 40 min following a color change of the amplified products. The sensitivity and specificity of the dry LAMP assay while using 152 DNA samples extracted from MGIT with confirmed results were 98% and 88%, respectively. When 43 randomly selected crude DNA samples from lesions were used, the sensitivity and specificity of the dry LAMP assay were 100% and 75%, respectively. Our LAMP assay offers the potential to meet the demands for a low-cost and rapid field detection tool for bTB in resource-limited countries in which bTB is endemic.
缺乏快速、准确和低成本的检测方法,阻碍了资源有限、疾病负担沉重的国家对牛结核病(bTB)采取积极的控制策略。我们开发了一种干式环介导等温扩增(LAMP)检测方法,用于快速和特异性检测bTB 的主要病原体 ,并使用在马拉维主要地区屠宰场进行常规肉品检验时收集的疑似 bTB 样本评估了该检测方法的效果。模板基因组 DNA 直接从肉芽肿性 bTB 样病变(粗提取 DNA)中提取,以及从孵育的分枝杆菌生长指示剂管(MGIT)中提取。在扩增产物发生颜色变化后 40 分钟内,通过肉眼即可观察到现场结果。当使用从 MGIT 中提取的 152 份经确认 结果的 DNA 样本进行检测时,干式 LAMP 检测方法的灵敏度和特异性分别为 98%和 88%。当随机选择 43 个病变的粗 DNA 样本进行检测时,干式 LAMP 检测方法的灵敏度和特异性分别为 100%和 75%。我们的 LAMP 检测方法有可能满足资源有限、bTB 流行的国家对低成本、快速现场检测 bTB 工具的需求。