Development of a real-time loop-mediated isothermal amplification assay for detection of Burkholderia mallei

作者：Pal V; Saxena A; Singh S; Goel A K; Kumar J S; Parida M M; Rai G P

来源：Transboundary and Emerging Diseases, 2018, 65(1): e32-e39.

DOI：10.1111/tbed.12658

• Burkholderia mallei
• glanders
• isothermal DNA amplification
• LAMP assay

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 1pg of B. mallei genomic DNA and 5.5x10(3)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.

• 出版日期2018-2