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Research Note: Rapid detection of avian infectious laryngotracheitis virus with real-time fluorescence-based recombinase-aided amplification.
- Source :
-
Poultry science [Poult Sci] 2020 Oct; Vol. 99 (10), pp. 4809-4813. Date of Electronic Publication: 2020 Jul 02. - Publication Year :
- 2020
-
Abstract
- In this study, specific primers and fluorescent probes were designed to target the thymidine kinase (TK) gene sequence of avian infectious laryngotracheitis virus (ILTV). Through specificity and sensitivity tests, a real-time fluorescence-based recombinase-aided amplification (RF-RAA) method for detecting ILTV was established. The results showed that the method was specific and could be used to accurately detect ILTV, and there was no cross-reaction with Newcastle disease virus (NDV), avian influenza virus (AIV), or infectious bronchitis virus (IBV). Real-time fluorescence-based recombinase-aided amplification had high sensitivity, and the lowest detectable limit (LDL) for ILTV could reach 10 copies/μL, 1,000 times more sensitive than conventional PCR (10 <superscript>4</superscript> copies/μL), to rival that of real-time fluorescence-based quantitative PCR (RFQ-PCR) (10 copies/μL). This method and RFQ-PCR were used to detect 96 samples of chicken throat swabs with ILT initially diagnosed in clinic from the north of China, and the coincidence rate of the 2 methods was 100%. The RF-RAA reaction required only 20-30 minutes to completing, and its sensitivity was much higher than that of conventional PCR. Real-time fluorescence-based recombinase-aided amplification is similar to RFQ-PCR and has the advantages of specificity, sensitivity, and high efficiency, so it is suitable for early clinical detection and epidemiological investigation of ILTV.<br /> (Copyright © 2020. Published by Elsevier Inc.)
- Subjects :
- Animals
Chickens
China
Fluorescence
Limit of Detection
Nucleic Acid Amplification Techniques veterinary
Recombinases metabolism
Sensitivity and Specificity
Herpesviridae Infections diagnosis
Herpesviridae Infections veterinary
Herpesviridae Infections virology
Herpesvirus 1, Gallid genetics
Poultry Diseases diagnosis
Poultry Diseases virology
Subjects
Details
- Language :
- English
- ISSN :
- 1525-3171
- Volume :
- 99
- Issue :
- 10
- Database :
- MEDLINE
- Journal :
- Poultry science
- Publication Type :
- Academic Journal
- Accession number :
- 32988516
- Full Text :
- https://doi.org/10.1016/j.psj.2020.06.025