title: Multiplex Amplification Refractory Mutation System PCR (ARMS-PCR) provides sequencing independent typing of canine parvovirus.
abstract: Canine parvovirus-2 antigenic variants (CPV-2a, CPV-2b and CPV-2c) ubiquitously distributed worldwide in canine population causes severe fatal gastroenteritis. Antigenic typing of CPV-2 remains a prime focus of research groups worldwide in understanding the disease epidemiology and virus evolution. The present study was thus envisioned to provide a simple sequencing independent, rapid, robust, specific, user-friendly technique for detecting and typing of presently circulating CPV-2 antigenic variants. ARMS-PCR strategy was employed using specific primers for CPV-2a, CPV-2b and CPV-2c to differentiate these antigenic types. ARMS-PCR was initially optimized with reference positive controls in two steps; where first reaction was used to differentiate CPV-2a from CPV-2b/CPV-2c. The second reaction was carried out with CPV-2c specific primers to confirm the presence of CPV-2c. Initial validation of the ARMS-PCR was carried out with 24 sequenced samples and the results were matched with the sequencing results. ARMS-PCR technique was further used to screen and type 90 suspected clinical samples. Randomly selected 15 suspected clinical samples that were typed with this technique were sequenced. The results of ARMS-PCR and the sequencing matched exactly with each other. The developed technique has a potential to become a sequencing independent method for simultaneous detection and typing of CPV-2 antigenic variants in veterinary disease diagnostic laboratories globally.
authors.affiliation: ICAR-Indian Veterinary Research Institute, Izatnagar, Bareilly, UP 243122, India.
authors.forename: Vishal
authors.initials: V
authors.lastname: Chander
authors.affiliation: ICAR-Indian Veterinary Research Institute, Izatnagar, Bareilly, UP 243122, India. Electronic address: soumendu.biotech@gmail.com.
authors.forename: Soumendu
authors.initials: S
authors.lastname: Chakravarti
authors.affiliation: ICAR-Indian Veterinary Research Institute, Izatnagar, Bareilly, UP 243122, India.
authors.forename: Vikas
authors.initials: V
authors.lastname: Gupta
authors.affiliation: ICAR-Indian Veterinary Research Institute, Izatnagar, Bareilly, UP 243122, India.
authors.forename: Sukdeb
authors.initials: S
authors.lastname: Nandi
authors.affiliation: ICAR-Indian Veterinary Research Institute, Izatnagar, Bareilly, UP 243122, India.
authors.forename: Mithilesh
authors.initials: M
authors.lastname: Singh
authors.affiliation: ICAR-Indian Veterinary Research Institute, Izatnagar, Bareilly, UP 243122, India.
authors.forename: Surendra Kumar
authors.initials: SK
authors.lastname: Badasara
authors.affiliation: ICAR-Indian Veterinary Research Institute, Izatnagar, Bareilly, UP 243122, India.
authors.forename: Chhavi
authors.initials: C
authors.lastname: Sharma
authors.affiliation: Defence Institute of Bio-Energy Research, Project Site Secunderabad, Military Farm Road, Old Bowenpally, Secunderabad, TS 500 011, India.
authors.forename: Mitesh
authors.initials: M
authors.lastname: Mittal
authors.affiliation: ICAR-Indian Veterinary Research Institute, Izatnagar, Bareilly, UP 243122, India. Electronic address: satyadandapat2008@hotmail.com.
authors.forename: S
authors.initials: S
authors.lastname: Dandapat
authors.affiliation: ICAR-Indian Veterinary Research Institute, Izatnagar, Bareilly, UP 243122, India.
authors.forename: V K
authors.initials: VK
authors.lastname: Gupta
mesh_terms: D000818:Animals; D000940:Antigenic Variation / Q000235:genetics; D004283:Dog Diseases / Q000821:virology; D004285:Dogs; D005243:Feces / Q000821:virology; D058889:Molecular Typing / Q000379:methods*; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D010322:Parvoviridae Infections / Q000662:veterinary / Q000821:virology; D017993:Parvovirus, Canine / Q000145:classification* / Q000235:genetics*
keywords: ARMS-PCR; Antigenic typing; Canine parvovirus