Veterinary Research Forum
Vet Res Forum

An accurate, rapid and simple loop-mediated isothermal amplification ‎method for Explanatum explanatum detection in animals

Document Type : Short Communication

Authors

Muhammad Rashid 1
Muhammad Imran Rashid 2
Qasim Ali 2

1 Dr. Panjwani Center for Molecular Medicine and Drug Research, International Center for Chemical and Biological Sciences, University of ‎Karachi, Karachi, Pakistan‎

2 Department of Parasitology, Faculty of Veterinary Science, University of Veterinary and Animal Sciences, Lahore, Pakistan

https://doi.org/10.30466/vrf.2020.111872.2657
Abstract
Paramphistomosis is an infectious disease caused by the liver flukes and it is associated with heavy loss of ruminant's population. Explanatum explanatum is a digenetic trematode commonly affecting domesticated ruminants. The available methods for pathogen detection are laborious and expensive and offer limited specificity; thus, considered not suitable for post mortem pathogen detection, surveillance and prevalence studies. New detection techniques offering simplicity, specificity and rapidity are absolutely needed. We have designed a loop-mediated isothermal amplification (LAMP) based polymerase chain reaction method, targeting a sequence of the explanatum species, using a primer pair from the internal transcribed spacer-2 region. The DNA from adult flukes belonging to explanatum species was isolated from infected livers and used to optimize the LAMP assay. The specificity and sensitivity of the LAMP assay were evaluated and found highly efficient in species-specific DNA detection with the sensitivity to detect 50.00 pg DNA in a 25.00 µL reaction mix. The procedure has the potential to be adapted for stool samples for field detection and disease surveillance/prevalence in rural and unprivileged areas.

Keywords

Explanatum explanatum
Liver flukes
‎ Loop-mediated isothermal ‎amplification
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Veterinary Research Forum
Volume 12, Issue 3
Summer 2021
Pages 377-381

  • Receive Date 24 July 2019
  • Revise Date 17 December 2019
  • Accept Date 05 January 2020

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