Comparison of a TaqMan real-time polymerase chain reaction assay with a loop-mediated isothermal amplification assay for detection of Gallid herpesvirus 1

Journal of Veterinary Diagnostic Investigation : Official Publication of the American Association of Veterinary Laboratory Diagnosticians, Inc
Shan-Chia OuKenneth S Macklin

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.

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Citations

Jan 28, 2014·Avian Pathology : Journal of the W.V.P.A·Kimberly R MenendezNathaniel L Tablante
Nov 1, 2013·World Journal of Virology·Shan-Chia Ou, Joseph J Giambrone
Nov 27, 2015·Journal of Water and Health·Kayo BiancoMaysa Mandetta Clementino
Dec 21, 2013·PloS One·Alessandra TammaroMark C Dessing
Oct 3, 2018·Journal of Environmental Quality·Asli AslanAsheley Chapman
Nov 13, 2013·Journal of Environmental Quality·Lisa M Durso
Dec 29, 2020·The Analyst·Mohamed El-TholothHaim H Bau

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Datasets Mentioned

BETA
EU104908

Methods Mentioned

BETA
PCR
electron microscopy
electrophoresis

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