Research Note: Rapid detection of avian infectious laryngotracheitis virus with real-time fluorescence-based recombinase-aided amplification
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 det...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2020-10-01
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| Series: | Poultry Science |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S0032579120303874 |
| _version_ | 1818851171539877888 |
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| author | Wenjing Wang Chunguang Wang Zichuang Zhang Peng Zhang Shanshan Yao Jingru Liu Xianghe Zhai Tie Zhang |
| author_facet | Wenjing Wang Chunguang Wang Zichuang Zhang Peng Zhang Shanshan Yao Jingru Liu Xianghe Zhai Tie Zhang |
| author_sort | Wenjing Wang |
| collection | DOAJ |
| description | 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 (104 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. |
| first_indexed | 2024-12-19T07:00:47Z |
| format | Article |
| id | doaj.art-fcee207d7db843a586c572b109af1233 |
| institution | Directory Open Access Journal |
| issn | 0032-5791 |
| language | English |
| last_indexed | 2024-12-19T07:00:47Z |
| publishDate | 2020-10-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Poultry Science |
| spelling | doaj.art-fcee207d7db843a586c572b109af12332022-12-21T20:31:26ZengElsevierPoultry Science0032-57912020-10-01991048094813Research Note: Rapid detection of avian infectious laryngotracheitis virus with real-time fluorescence-based recombinase-aided amplificationWenjing Wang0Chunguang Wang1Zichuang Zhang2Peng Zhang3Shanshan Yao4Jingru Liu5Xianghe Zhai6Tie Zhang7College of Veterinary Medicine, Hebei Agricultural University, Baoding 071001, ChinaCollege of Veterinary Medicine, Hebei Agricultural University, Baoding 071001, ChinaCollege of Animal Science and Technology, Jiangxi Agricultural University, Nanchang 330045, ChinaCollege of Veterinary Medicine, Hebei Agricultural University, Baoding 071001, ChinaCollege of Veterinary Medicine, Hebei Agricultural University, Baoding 071001, ChinaCollege of Veterinary Medicine, Hebei Agricultural University, Baoding 071001, ChinaCollege of Veterinary Medicine, Hebei Agricultural University, Baoding 071001, ChinaCollege of Veterinary Medicine, Hebei Agricultural University, Baoding 071001, China; Corresponding author: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 (104 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.http://www.sciencedirect.com/science/article/pii/S0032579120303874infectious laryngotracheitis virusreal-time fluorescence-based recombinase-aided amplificationTK gene |
| spellingShingle | Wenjing Wang Chunguang Wang Zichuang Zhang Peng Zhang Shanshan Yao Jingru Liu Xianghe Zhai Tie Zhang Research Note: Rapid detection of avian infectious laryngotracheitis virus with real-time fluorescence-based recombinase-aided amplification Poultry Science infectious laryngotracheitis virus real-time fluorescence-based recombinase-aided amplification TK gene |
| title | Research Note: Rapid detection of avian infectious laryngotracheitis virus with real-time fluorescence-based recombinase-aided amplification |
| title_full | Research Note: Rapid detection of avian infectious laryngotracheitis virus with real-time fluorescence-based recombinase-aided amplification |
| title_fullStr | Research Note: Rapid detection of avian infectious laryngotracheitis virus with real-time fluorescence-based recombinase-aided amplification |
| title_full_unstemmed | Research Note: Rapid detection of avian infectious laryngotracheitis virus with real-time fluorescence-based recombinase-aided amplification |
| title_short | Research Note: Rapid detection of avian infectious laryngotracheitis virus with real-time fluorescence-based recombinase-aided amplification |
| title_sort | research note rapid detection of avian infectious laryngotracheitis virus with real time fluorescence based recombinase aided amplification |
| topic | infectious laryngotracheitis virus real-time fluorescence-based recombinase-aided amplification TK gene |
| url | http://www.sciencedirect.com/science/article/pii/S0032579120303874 |
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