Document Type : Short Communication
Dr.Panjwani Center for Molecular Medicine and Drug Research PCMD, Store, University of Karachi
Professor, Department of Parasitology University of Veterinary and Animal Sciences, UVAS Syed Abdul Qadir Jillani (Out Fall) Road, Lahore 54200- Pakistan
Department of Pathobiology Gomal University: Dera Ismail Khan, KPK
Paramphistomosis is an infectious disease, caused by the liver flukes, and 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, expensive and offer limited specificity, thus considered not suitable for postmortem pathogen detection, surveillance and prevalence studies. New detection techniques which offer simplicity, specificity and rapidity are absolutely needed. We have designed a Loop-Mediated Isothermal Amplification (LAMP) based PCR method, targeting a sequence of the explanatum species, using a primer pair from the internal transcribed spacer-2 (ITS2) region. DNA from adult flukes belonging to species explanatum 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 detecting species-specific DNA with the sensitivity to detect 50pg DNA in a 25 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.