Microbiology Division, Defence Research and Development Establishment, Gwalior, India.
Biotechnology Division, Defence Research and Development Establishment, Gwalior, India.
Transbound Emerg Dis. 2018 Feb;65(1):e32-e39. doi: 10.1111/tbed.12665. Epub 2017 Jun 25.
Burkholderia mallei is the aetiological agent of glanders, a highly contagious and re-emerging zoonotic disease. Early diagnosis of glanders is critically important to ensure timely treatment with appropriate antibiotics in humans, and to prevent spread of infection in animals. Molecular detection of B. mallei has always been troublesome because of its genetic similarity with Burkholderia pseudomallei, the causative agent of melioidosis. In present investigation, a set of six B. mallei-specific primers were designed and a simple, rapid, specific and sensitive real-time loop-mediated isothermal amplification (LAMP) assay was developed for detection of B. mallei. The LAMP assay could detect as low as 1 pg of B. mallei genomic DNA and 5.5 × 10 CFU/ml of B. mallei in spiked human blood. The assay was highly specific for B. mallei as it did not cross-react with other bacterial strains used in the study. The established LAMP assay is field adaptable and can be a better and viable alternative to PCR-based techniques for detection of B. mallei in glanders endemic areas with resource-limited settings.
鼻疽伯克霍尔德菌是马鼻疽的病原体，马鼻疽是一种高度传染性和重现的人畜共患病。早期诊断马鼻疽对于确保人类及时使用适当的抗生素进行治疗以及防止动物感染传播至关重要。由于其与鼻疽假单胞菌（导致类鼻疽的病原体）的遗传相似性，鼻疽伯克霍尔德菌的分子检测一直存在困难。在本研究中，设计了一组 6 个鼻疽伯克霍尔德菌特异性引物，并开发了一种简单、快速、特异和敏感的实时环介导等温扩增（LAMP）检测方法。LAMP 检测可检测低至 1pg 的鼻疽伯克霍尔德菌基因组 DNA 和 5.5×10 个 CFU/ml 的人工污染人血中的鼻疽伯克霍尔德菌。该检测方法对鼻疽伯克霍尔德菌具有高度特异性，因为它与研究中使用的其他细菌菌株没有交叉反应。建立的 LAMP 检测方法适用于现场，并且可以作为基于 PCR 技术的更好和可行的替代方法，用于在资源有限的地方性鼻疽流行地区检测鼻疽伯克霍尔德菌。
