title: A double-stranded probe coupled with isothermal amplification for qualitative and quantitative detection of avian reovirus.
abstract: We applied a probe-based real-time loop-mediated isothermal amplification (Cy5-RTqLAMP) technique targeting the avian reovirus (ARV) S3 gene to develop a rapid, sensitive, and specific method for virus detection and quantification. This test specifically detected the presence of ARV, but not other viruses or bacteria present in clinical or artificially spiked samples, including Newcastle disease virus, infectious bursal disease virus, fowl adenovirus, Marek's disease virus, Escherichia coli, and Salmonella spp. This test can detect ARV in less than one hour with an analytical sensitivity of 10 viral gene copies and 1 fg of total cDNA. The Cy5-RTqLAMP does not yield false positive results and is 100 times more sensitive than conventional PCR. This test was shown to be able to detect the presence of ARV in clinical samples. A similar strategy may be used for detection of other important human and animal viral pathogens.
authors.affiliation: Division of Veterinary Biotechnology, Indian Veterinary Research Institute, Izatnagar, 243122, UP, India. deep_biotek@yahoo.com.
authors.forename: D
authors.initials: D
authors.lastname: Kumar
authors.affiliation: Division of Veterinary Biotechnology, Indian Veterinary Research Institute, Izatnagar, 243122, UP, India.
authors.forename: T K S
authors.initials: TK
authors.lastname: Chauhan
authors.affiliation: Division of Livestock Product Technology, Indian Veterinary Research Institute, Izatnagar, 243122, UP, India.
authors.forename: R K
authors.initials: RK
authors.lastname: Agarwal
authors.affiliation: Avian Disease Section, Indian Veterinary Research Institute, Izatnagar, 243122, UP, India.
authors.forename: K
authors.initials: K
authors.lastname: Dhama
authors.affiliation: Division of Veterinary Biotechnology, Indian Veterinary Research Institute, Izatnagar, 243122, UP, India.
authors.forename: P P
authors.initials: PP
authors.lastname: Goswami
authors.affiliation: Avian Disease Section, Indian Veterinary Research Institute, Izatnagar, 243122, UP, India.
authors.forename: A K
authors.initials: AK
authors.lastname: Mariappan
authors.affiliation: Division of Veterinary Biotechnology, Indian Veterinary Research Institute, Izatnagar, 243122, UP, India.
authors.forename: A K
authors.initials: AK
authors.lastname: Tiwari
authors.affiliation: Division of Veterinary Biotechnology, Indian Veterinary Research Institute, Izatnagar, 243122, UP, India.
authors.forename: B P
authors.initials: BP
authors.lastname: Mishra
mesh_terms: D000818:Animals; D002645:Chickens; D017931:DNA Primers / Q000235:genetics; D004279:DNA, Viral / Q000235:genetics*; D006801:Humans; D021141:Nucleic Acid Amplification Techniques / Q000295:instrumentation / Q000379:methods*; D030101:Orthoreovirus, Avian / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000295:instrumentation / Q000379:methods; D011201:Poultry Diseases / Q000175:diagnosis / Q000821:virology*; D012088:Reoviridae Infections / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology
chemical_list: D017931:DNA Primers; D004279:DNA, Viral
keywords: Avian reovirus; Diagnosis; Loop-mediated isothermal amplification (LAMP); S3 gene; Virus copy number