Development and application of a triplex real-time PCR assay for the detection of H3, H4, and H5 subtypes of avian influenza virus
ABSTRACT: Avian influenza virus (AIV) poses a significant threat to the poultry industry and public health. Among the diverse AIV subtypes, H3, H4, and H5 are frequently detected in waterfowl and live poultry markets (LPM). The expeditious and precise identification of these subtypes is imperative i...
| Main Authors: | , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2024-02-01
|
| Series: | Poultry Science |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S0032579123008532 |
| _version_ | 1797317440431980544 |
|---|---|
| author | Feng Wen Congying Wang Jinyue Guo Hai Yu Sheng Yuan Yong Li Zhili Li Shujian Huang Zhaoping Liang |
| author_facet | Feng Wen Congying Wang Jinyue Guo Hai Yu Sheng Yuan Yong Li Zhili Li Shujian Huang Zhaoping Liang |
| author_sort | Feng Wen |
| collection | DOAJ |
| description | ABSTRACT: Avian influenza virus (AIV) poses a significant threat to the poultry industry and public health. Among the diverse AIV subtypes, H3, H4, and H5 are frequently detected in waterfowl and live poultry markets (LPM). The expeditious and precise identification of these subtypes is imperative in impeding the dissemination of the disease. In this study, we have developed a triplex real-time PCR assay endowed with the capacity to simultaneously discriminate AIV subtypes H3, H4, and H5. This method showcases remarkable specificity, selectively amplifying H3, H4, and H5 AIV subtypes sans any cross-reactivity with other subtypes or common avian pathogens. Furthermore, this method exhibits high sensitivity, with a detection threshold of 2.1 × 102 copies/μL for H3, H4, and H5 AIV subtypes. Additionally, the assay demonstrates reproducibility, as evidenced by intra- and interassay variability, with a coefficient of variation below 1.5%. A total of 338 cloacal swabs were collected from LPM to evaluate the performance of our assay. The obtained results evinced a high level of concordance with the sequencing data. In summary, our study has developed a triplex real-time PCR method that can be employed in laboratory-based testing and surveillance of AIV. This assay holds promise in augmenting our ability to detect and monitor AIV subtypes, thereby facilitating timely interventions and safeguarding both the poultry industry and public health. |
| first_indexed | 2024-03-08T03:36:00Z |
| format | Article |
| id | doaj.art-35ad787d26ee41b189433c6d27084cd8 |
| institution | Directory Open Access Journal |
| issn | 0032-5791 |
| language | English |
| last_indexed | 2024-03-08T03:36:00Z |
| publishDate | 2024-02-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Poultry Science |
| spelling | doaj.art-35ad787d26ee41b189433c6d27084cd82024-02-10T04:43:42ZengElsevierPoultry Science0032-57912024-02-011032103333Development and application of a triplex real-time PCR assay for the detection of H3, H4, and H5 subtypes of avian influenza virusFeng Wen0Congying Wang1Jinyue Guo2Hai Yu3Sheng Yuan4Yong Li5Zhili Li6Shujian Huang7Zhaoping Liang8College of Life Science and Engineering, Foshan University, Foshan 528231, Guangdong, ChinaCollege of Life Science and Engineering, Foshan University, Foshan 528231, Guangdong, ChinaCollege of Life Science and Engineering, Foshan University, Foshan 528231, Guangdong, ChinaShanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai 200241, ChinaCollege of Life Science and Engineering, Foshan University, Foshan 528231, Guangdong, ChinaCollege of Animal Science and Technology, Jiangxi Agricultural University, Nanchang 330045, Jiangxi, ChinaCollege of Life Science and Engineering, Foshan University, Foshan 528231, Guangdong, ChinaCollege of Life Science and Engineering, Foshan University, Foshan 528231, Guangdong, ChinaCollege of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, Guangdong, China; Corresponding author:ABSTRACT: Avian influenza virus (AIV) poses a significant threat to the poultry industry and public health. Among the diverse AIV subtypes, H3, H4, and H5 are frequently detected in waterfowl and live poultry markets (LPM). The expeditious and precise identification of these subtypes is imperative in impeding the dissemination of the disease. In this study, we have developed a triplex real-time PCR assay endowed with the capacity to simultaneously discriminate AIV subtypes H3, H4, and H5. This method showcases remarkable specificity, selectively amplifying H3, H4, and H5 AIV subtypes sans any cross-reactivity with other subtypes or common avian pathogens. Furthermore, this method exhibits high sensitivity, with a detection threshold of 2.1 × 102 copies/μL for H3, H4, and H5 AIV subtypes. Additionally, the assay demonstrates reproducibility, as evidenced by intra- and interassay variability, with a coefficient of variation below 1.5%. A total of 338 cloacal swabs were collected from LPM to evaluate the performance of our assay. The obtained results evinced a high level of concordance with the sequencing data. In summary, our study has developed a triplex real-time PCR method that can be employed in laboratory-based testing and surveillance of AIV. This assay holds promise in augmenting our ability to detect and monitor AIV subtypes, thereby facilitating timely interventions and safeguarding both the poultry industry and public health.http://www.sciencedirect.com/science/article/pii/S0032579123008532avian influenza virusH3H4H5TaqMan probe |
| spellingShingle | Feng Wen Congying Wang Jinyue Guo Hai Yu Sheng Yuan Yong Li Zhili Li Shujian Huang Zhaoping Liang Development and application of a triplex real-time PCR assay for the detection of H3, H4, and H5 subtypes of avian influenza virus Poultry Science avian influenza virus H3 H4 H5 TaqMan probe |
| title | Development and application of a triplex real-time PCR assay for the detection of H3, H4, and H5 subtypes of avian influenza virus |
| title_full | Development and application of a triplex real-time PCR assay for the detection of H3, H4, and H5 subtypes of avian influenza virus |
| title_fullStr | Development and application of a triplex real-time PCR assay for the detection of H3, H4, and H5 subtypes of avian influenza virus |
| title_full_unstemmed | Development and application of a triplex real-time PCR assay for the detection of H3, H4, and H5 subtypes of avian influenza virus |
| title_short | Development and application of a triplex real-time PCR assay for the detection of H3, H4, and H5 subtypes of avian influenza virus |
| title_sort | development and application of a triplex real time pcr assay for the detection of h3 h4 and h5 subtypes of avian influenza virus |
| topic | avian influenza virus H3 H4 H5 TaqMan probe |
| url | http://www.sciencedirect.com/science/article/pii/S0032579123008532 |
| work_keys_str_mv | AT fengwen developmentandapplicationofatriplexrealtimepcrassayforthedetectionofh3h4andh5subtypesofavianinfluenzavirus AT congyingwang developmentandapplicationofatriplexrealtimepcrassayforthedetectionofh3h4andh5subtypesofavianinfluenzavirus AT jinyueguo developmentandapplicationofatriplexrealtimepcrassayforthedetectionofh3h4andh5subtypesofavianinfluenzavirus AT haiyu developmentandapplicationofatriplexrealtimepcrassayforthedetectionofh3h4andh5subtypesofavianinfluenzavirus AT shengyuan developmentandapplicationofatriplexrealtimepcrassayforthedetectionofh3h4andh5subtypesofavianinfluenzavirus AT yongli developmentandapplicationofatriplexrealtimepcrassayforthedetectionofh3h4andh5subtypesofavianinfluenzavirus AT zhilili developmentandapplicationofatriplexrealtimepcrassayforthedetectionofh3h4andh5subtypesofavianinfluenzavirus AT shujianhuang developmentandapplicationofatriplexrealtimepcrassayforthedetectionofh3h4andh5subtypesofavianinfluenzavirus AT zhaopingliang developmentandapplicationofatriplexrealtimepcrassayforthedetectionofh3h4andh5subtypesofavianinfluenzavirus |