Comparison of a TaqMan real-time polymerase chain reaction assay with a loop-mediated isothermal amplification assay for detection of Gallid herpesvirus 1
| Autor: | Joseph J. Giambrone, Shan-Chia Ou, Kenneth S. Macklin |
|---|---|
| Rok vydání: | 2011 |
| Předmět: |
Detection limit
Gallid herpesvirus 1 General Veterinary biology Chemistry Loop-mediated isothermal amplification Recombinase Polymerase Amplification Herpesviridae Infections Real-Time Polymerase Chain Reaction biology.organism_classification Sensitivity and Specificity Virology Molecular biology law.invention Titer Real-time polymerase chain reaction Herpesvirus 1 Gallid law TaqMan Animals Chickens Nucleic Acid Amplification Techniques Poultry Diseases Polymerase chain reaction |
| Zdroj: | Journal of Veterinary Diagnostic Investigation. 24:138-141 |
| ISSN: | 1943-4936 1040-6387 |
| DOI: | 10.1177/1040638711427578 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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