title: Comparison of a TaqMan real-time polymerase chain reaction assay with a loop-mediated isothermal amplification assay for detection of Gallid herpesvirus 1.
abstract: A TaqMan real-time polymerase chain reaction (PCR) and loop-mediated isothermal amplification (LAMP) assay were developed to detect Gallid herpesvirus 1 (GaHV-1, formerly Infectious laryngotracheitis virus). The standard curve of real-time PCR was established, and the sensitivity reached 10 copies/μl. In the current study, the conversion between viral titer and GaHV-1 genomic copy number was constructed. Six primers for LAMP assay amplified target gene at 65°C within 45 min, and the detection limit was 60 copies/μl. The 6 primers were highly specific, sensitive, and reproducible for detection of GaHV-1. Although the sensitivity of LAMP was lower than that of real-time PCR, LAMP was faster, less expensive, and did not require a thermocycler. The LAMP assay would be a viable alternative assay in diagnostic laboratories that do not employ real-time PCR technology.
authors.affiliation: Poultry Science Department, Auburn University, Auburn, AL 36849-5416, USA.
authors.forename: Shan-Chia
authors.initials: SC
authors.lastname: Ou
authors.forename: Joseph J
authors.initials: JJ
authors.lastname: Giambrone
authors.forename: Kenneth S
authors.initials: KS
authors.lastname: Macklin
mesh_terms: D000818:Animals; D002645:Chickens / Q000821:virology; D006566:Herpesviridae Infections / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology; D001352:Herpesvirus 1, Gallid / Q000235:genetics*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods / Q000662:veterinary*; D011201:Poultry Diseases / Q000175:diagnosis / Q000821:virology*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D012680:Sensitivity and Specificity