title: Development and evaluation of loop-mediated isothermal amplification assay for rapid detection of Capripoxvirus.
abstract: OBJECTIVE
  The present study was undertaken to develop a nucleic acid-based diagnostic assay loop-mediated isothermal amplification assay (LAMP) targeting highly conserved genomic regions of Capripoxvirus (CaPVs) and its comparative evaluation with real-time polymerase chain reaction (PCR).
  
  
  METHODS
  Lyophilized vaccine strain of sheeppox virus (SPPV) was used for optimization of LAMP assay. The LAMP assay was designed using envelope immunogenic protein (P32) coding gene targeting highly conserved genomic regions of CaPV responsible for causing sheep pox, goat pox, and lumpy skin disease in sheep, goat and cattle respectively. Serial tenfold dilution of SPPV recombinant plasmid DNA was used for a calculating limit of detection. Analytical sensitivity and specificity were performed.
  
  
  RESULTS
  The test described is quick (30 min), sensitive and specific for detection of CaPVs. The described assay did not show any cross-reactivity to other related viruses that cause apparently similar clinical signs. It was found to be ten times more sensitive than conventional PCR however, 100 times less sensitive than quantitative PCR (qPCR). LAMP assay results were monitored by color change method using picogreen dye and agarose gel electrophoresis.
  
  
  CONCLUSIONS
  LAMP assay can be a very good alternative for CaPV detection to other molecular techniques requiring sophisticated equipments.
authors.affiliation: Department of Animal Biotechnology, College of Veterinary Sciences, LLR University of Veterinary and Animal Sciences, Hisar, Haryana, India.
authors.forename: Kanisht
authors.initials: K
authors.lastname: Batra
authors.affiliation: Department of Animal Biotechnology, College of Veterinary Sciences, LLR University of Veterinary and Animal Sciences, Hisar, Haryana, India.
authors.forename: Aman
authors.initials: A
authors.lastname: Kumar
authors.affiliation: Department of Animal Biotechnology, College of Veterinary Sciences, LLR University of Veterinary and Animal Sciences, Hisar, Haryana, India.
authors.forename: Vinay
authors.initials: V
authors.lastname: Kumar
authors.affiliation: Department of Animal Biotechnology, College of Veterinary Sciences, LLR University of Veterinary and Animal Sciences, Hisar, Haryana, India.
authors.forename: Trilok
authors.initials: T
authors.lastname: Nanda
authors.affiliation: Resource Faculty, Department of Animal Biotechnology, College of Veterinary Sciences, Lala Lajpat Rai University of Veterinary and Animal Sciences, Hisar, Haryana, India.
authors.forename: Narender S
authors.initials: NS
authors.lastname: Maan
authors.affiliation: Department of Animal Biotechnology, College of Veterinary Sciences, LLR University of Veterinary and Animal Sciences, Hisar, Haryana, India.
authors.forename: Sushila
authors.initials: S
authors.lastname: Maan
keywords: Capripoxvirus; loop-mediated isothermal amplification assay; real-time polymerase chain reaction; sensitivity; specificity