Bioprocess Technology Division, Defence Research and Development Establishment, Gwalior, India.
Transbound Emerg Dis. 2019 Mar;66(2):1016-1022. doi: 10.1111/tbed.13126. Epub 2019 Feb 15.
Burkholderia mallei, a potential biothreat agent is the aetiological agent of glanders, a zoonotic disease primarily affecting equines. B. mallei shares close genetic proximity with B. pseudomallei, the aetiological agent of melioidosis. Hence, molecular detection of B. mallei and its differentiation from B. pseudomallei has always been challenging. Early diagnosis of glanders is critical for timely treatment in humans and disease containment in animals. In this study a recombinase polymerase amplification-lateral flow (RPA-LF) assay has been developed for early and accurate detection of B. mallei. RPA-LF assay was found to be highly sensitive and detected as low as 10 fg genomic DNA of B. mallei. The assay was highly specific and could differentiate B. mallei and B. pseudomallei. The assay also detected B. mallei in artificially spiked blood, tap water and garden soil. The established assay is simple, rapid and does not require complex instrumentation. The field deployable assay can have better implications in rapid glanders diagnosis and environmental detection of B. mallei over PCR-based detection tools in glanders endemic areas with limited laboratory resources.
马鼻疽伯克霍尔德菌是一种潜在的生物威胁因子，是马传染性鼻疽的病原体，主要感染马属动物。马鼻疽伯克霍尔德菌与类鼻疽伯克霍尔德菌亲缘关系密切，后者是类鼻疽病的病原体。因此，马鼻疽伯克霍尔德菌的分子检测及其与类鼻疽伯克霍尔德菌的区分一直具有挑战性。早期诊断鼻疽对人类的及时治疗和动物疫病的控制至关重要。本研究建立了一种重组酶聚合酶扩增-侧向流动（RPA-LF）检测方法，用于早期、准确地检测马鼻疽伯克霍尔德菌。该检测方法具有很高的灵敏度，最低可检测到 10 fg 的马鼻疽伯克霍尔德菌基因组 DNA。该检测方法具有高度特异性，可区分马鼻疽伯克霍尔德菌和类鼻疽伯克霍尔德菌。该检测方法还可检测人工污染血液、自来水和花园土壤中的马鼻疽伯克霍尔德菌。建立的检测方法简单、快速，不需要复杂的仪器。在鼻疽病流行地区，现场可部署的检测方法比基于 PCR 的检测工具在资源有限的实验室中具有更好的快速诊断鼻疽和环境中检测马鼻疽伯克霍尔德菌的应用前景。
