Development and evaluation of RT-qPCR assays for two neglected orthobunyaviruses: Oya virus and Ebinur Lake virus
Objectives: Oya virus (OYAV) and Ebinur lake virus (EBIV) belong to the genus Orthobunyavirus within the Peribunyaviridae family, and both are recognized as the novel virus with potential threat to the animal or public health. Given their potential to cause outbreaks and their detection in diverse s...
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| Format: | Article |
| Language: | English |
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Elsevier
2024-01-01
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| Series: | Virus Research |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S0168170223002277 |
| _version_ | 1797616003766550528 |
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| author | Siyuan Liu Wei Chen Raphael Nyaruaba Shunlong Wang Cihan Yang Qun Wu Ying Liu Puyu Liu Fei Wang Jingling Wang Zhiming Yuan Dingwei Sun Han Xia |
| author_facet | Siyuan Liu Wei Chen Raphael Nyaruaba Shunlong Wang Cihan Yang Qun Wu Ying Liu Puyu Liu Fei Wang Jingling Wang Zhiming Yuan Dingwei Sun Han Xia |
| author_sort | Siyuan Liu |
| collection | DOAJ |
| description | Objectives: Oya virus (OYAV) and Ebinur lake virus (EBIV) belong to the genus Orthobunyavirus within the Peribunyaviridae family, and both are recognized as the novel virus with potential threat to the animal or public health. Given their potential to cause outbreaks and their detection in diverse samples across different regions, the need for a reliable and efficient molecular detection method for OYAV and EBIV becomes imperative. Methods: The S-segment of OYAV and EBIV was used for designing specific primer and probe sets, which were employed in a real-time reverse transcription quantitative PCR (RT-qPCR) assay. The analytical performance of these assays, encompassing specificity, sensitivity, reproducibility, and fitness for purpose, was thoroughly evaluated across various sample matrices. Results: The developed RT-qPCR assays were very specific to their respective targets. Both assays were highly reproducible (%CV<3) and sensitive with the 95% limit of detection (LOD) of 0.80 PFU/mL for OYAV primer probe set and 0.37 PFU/mL for EBIV primer probe set. Furthermore, the assays fitness for purpose was good as it could detect the specific viruses in virus-spiked serum samples, virus-inoculated mosquito samples, field caught mosquitoes and biting midge samples. Conclusions: Our study has successfully developed specific, sensitive, and reliable RT-qPCR assays for the detection of OYAV and EBIV. These assays hold great promise for their potential application in clinical and field samples in the future. |
| first_indexed | 2024-03-11T07:34:58Z |
| format | Article |
| id | doaj.art-7d8a2a8f6cba4b4493e7797ffc2b035f |
| institution | Directory Open Access Journal |
| issn | 1872-7492 |
| language | English |
| last_indexed | 2024-03-11T07:34:58Z |
| publishDate | 2024-01-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Virus Research |
| spelling | doaj.art-7d8a2a8f6cba4b4493e7797ffc2b035f2023-11-17T05:24:56ZengElsevierVirus Research1872-74922024-01-01339199265Development and evaluation of RT-qPCR assays for two neglected orthobunyaviruses: Oya virus and Ebinur Lake virusSiyuan Liu0Wei Chen1Raphael Nyaruaba2Shunlong Wang3Cihan Yang4Qun Wu5Ying Liu6Puyu Liu7Fei Wang8Jingling Wang9Zhiming Yuan10Dingwei Sun11Han Xia12These authors contributed equally to this work.; Key Laboratory of Special Pathogens and Biosafety, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, Hubei, China; University of Chinese Academy of Sciences, Beijing, ChinaThese authors contributed equally to this work.; Key Laboratory of Special Pathogens and Biosafety, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, Hubei, China; University of Chinese Academy of Sciences, Beijing, ChinaKey Laboratory of Special Pathogens and Biosafety, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, Hubei, China; University of Chinese Academy of Sciences, Beijing, China; Sino-Africa Joint Research Center, Chinese Academy of Sciences, Wuhan, Hubei, ChinaKey Laboratory of Special Pathogens and Biosafety, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, Hubei, China; University of Chinese Academy of Sciences, Beijing, ChinaKey Laboratory of Special Pathogens and Biosafety, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, Hubei, China; University of Chinese Academy of Sciences, Beijing, ChinaKey Laboratory of Special Pathogens and Biosafety, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, Hubei, China; University of Chinese Academy of Sciences, Beijing, ChinaHainan Provincial Center for Disease Control and Prevention, Haikou, ChinaHainan Provincial Center for Disease Control and Prevention, Haikou, ChinaKey Laboratory of Special Pathogens and Biosafety, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, Hubei, ChinaYunnan Tropical and Subtropical Animal Virus Disease Laboratory, Yunnan Animal Science and Veterinary Institute, Kunming, Yunnan Province, ChinaKey Laboratory of Special Pathogens and Biosafety, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, Hubei, China; University of Chinese Academy of Sciences, Beijing, ChinaHainan Provincial Center for Disease Control and Prevention, Haikou, China; Corresponding author.Corresponding author.; Key Laboratory of Special Pathogens and Biosafety, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, Hubei, China; University of Chinese Academy of Sciences, Beijing, China; Hubei Jiangxia Laboratory, Wuhan, ChinaObjectives: Oya virus (OYAV) and Ebinur lake virus (EBIV) belong to the genus Orthobunyavirus within the Peribunyaviridae family, and both are recognized as the novel virus with potential threat to the animal or public health. Given their potential to cause outbreaks and their detection in diverse samples across different regions, the need for a reliable and efficient molecular detection method for OYAV and EBIV becomes imperative. Methods: The S-segment of OYAV and EBIV was used for designing specific primer and probe sets, which were employed in a real-time reverse transcription quantitative PCR (RT-qPCR) assay. The analytical performance of these assays, encompassing specificity, sensitivity, reproducibility, and fitness for purpose, was thoroughly evaluated across various sample matrices. Results: The developed RT-qPCR assays were very specific to their respective targets. Both assays were highly reproducible (%CV<3) and sensitive with the 95% limit of detection (LOD) of 0.80 PFU/mL for OYAV primer probe set and 0.37 PFU/mL for EBIV primer probe set. Furthermore, the assays fitness for purpose was good as it could detect the specific viruses in virus-spiked serum samples, virus-inoculated mosquito samples, field caught mosquitoes and biting midge samples. Conclusions: Our study has successfully developed specific, sensitive, and reliable RT-qPCR assays for the detection of OYAV and EBIV. These assays hold great promise for their potential application in clinical and field samples in the future.http://www.sciencedirect.com/science/article/pii/S0168170223002277Oya virus (OYAV)Ebinur lake virus (EBIV)RT-qPCRVirus detection |
| spellingShingle | Siyuan Liu Wei Chen Raphael Nyaruaba Shunlong Wang Cihan Yang Qun Wu Ying Liu Puyu Liu Fei Wang Jingling Wang Zhiming Yuan Dingwei Sun Han Xia Development and evaluation of RT-qPCR assays for two neglected orthobunyaviruses: Oya virus and Ebinur Lake virus Virus Research Oya virus (OYAV) Ebinur lake virus (EBIV) RT-qPCR Virus detection |
| title | Development and evaluation of RT-qPCR assays for two neglected orthobunyaviruses: Oya virus and Ebinur Lake virus |
| title_full | Development and evaluation of RT-qPCR assays for two neglected orthobunyaviruses: Oya virus and Ebinur Lake virus |
| title_fullStr | Development and evaluation of RT-qPCR assays for two neglected orthobunyaviruses: Oya virus and Ebinur Lake virus |
| title_full_unstemmed | Development and evaluation of RT-qPCR assays for two neglected orthobunyaviruses: Oya virus and Ebinur Lake virus |
| title_short | Development and evaluation of RT-qPCR assays for two neglected orthobunyaviruses: Oya virus and Ebinur Lake virus |
| title_sort | development and evaluation of rt qpcr assays for two neglected orthobunyaviruses oya virus and ebinur lake virus |
| topic | Oya virus (OYAV) Ebinur lake virus (EBIV) RT-qPCR Virus detection |
| url | http://www.sciencedirect.com/science/article/pii/S0168170223002277 |
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