Rapid diagnosis of Leishmania infection with a portable loopmediated isothermal amplification device

J Biosci. 2021;46(4):92. doi: 10.1007/s12038-021-00211-0.

Abstract

L. donovani is an intracellular protozoan parasite, that causes visceral leishmaniasis (VL), and consequently, post-kala azar dermal leishmaniasis (PKDL). Diagnosis and treatment of leishmaniasis is crucial for decreasing its transmission. Various diagnostic techniques like microscopy, enzyme-linked immunosorbent assays (ELISA) and PCR-based methods are used to detect leishmaniasis infection. More recently, loop-mediated isothermal amplification (LAMP) assay has emerged as an ideal diagnostic measure for leishmaniasis, primarily due to its accuracy, speed and simplicity. However, point-of-care diagnosis is still not been tested with the LAMP assay. We have developed a portable LAMP device for the monitoring of Leishmania infection. The LAMP assay performed using our device can detect and amplify as little as 100 femtograms of L. donovani DNA. In a preliminary study, we have shown that the device can also amplify L. donovani DNA present in VL and PKDL patient samples with high sensitivity (100%), specificity (98%) and accuracy (99%), and can be used both for diagnostic and prognostic analysis. To our knowledge, this is the first report to describe the development and application of a portable LAMP device which has the potential to evolve as a point-of-care diagnostic and prognostic tool for Leishmania infections in future.

MeSH terms

  • Case-Control Studies
  • DNA, Protozoan / genetics
  • Equipment Design
  • Fluorescence
  • Humans
  • Leishmania donovani / genetics
  • Leishmaniasis / diagnosis*
  • Leishmaniasis / parasitology*
  • Leprosy / parasitology
  • Molecular Diagnostic Techniques / instrumentation*
  • Molecular Diagnostic Techniques / methods
  • Nucleic Acid Amplification Techniques / instrumentation*
  • Nucleic Acid Amplification Techniques / methods
  • Parasite Load
  • Point-of-Care Systems
  • Polymerase Chain Reaction / methods
  • Sensitivity and Specificity

Substances

  • DNA, Protozoan

Supplementary concepts

  • LAMP assay