Replication of Human Norovirus in Human Intestinal Enteroids Is Affected by Fecal Sample Processing
Human intestinal enteroids (HIEs) culture is an emerging model for assessing the infectivity of human noroviruses (HuNoVs). The model is based on detecting an increase in HuNoV RNA post-infection of HIEs. HuNoV fecal samples used for HIE infection are traditionally processed by serial filtration. Re...
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| Format: | Article |
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MDPI AG
2024-02-01
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| Series: | Viruses |
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| Online Access: | https://www.mdpi.com/1999-4915/16/2/241 |
| _version_ | 1797296820029751296 |
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| author | Revati Narwankar Malak A. Esseili |
| author_facet | Revati Narwankar Malak A. Esseili |
| author_sort | Revati Narwankar |
| collection | DOAJ |
| description | Human intestinal enteroids (HIEs) culture is an emerging model for assessing the infectivity of human noroviruses (HuNoVs). The model is based on detecting an increase in HuNoV RNA post-infection of HIEs. HuNoV fecal samples used for HIE infection are traditionally processed by serial filtration. Recently, processing HuNoV fecal samples by serial centrifugation was shown to retain vesicles containing HuNoV. The objective of this study was to investigate whether serially centrifuged fecal samples, RNA extraction kit (QIAamp versus MagMaX) and HIE age (newer versus older) affect HuNoV RNA fold increase in HIE. HuNoV GII.1, GII.4 and GII.6 fecal samples were prepared by serial centrifugation and filtration and the viral RNA in HIE was quantified at 1 and 72 h post-infection (hpi) following RNA extraction and RT-qPCR. The serially filtered GII.1, GII.4 and GII.6 showed successful replication in HIE, resulting in mean log increases of 2.2, 2 and 1.2, respectively, at 72 vs. 1 hpi. In contrast, only serially centrifuged GII.1 showed consistently successful replication. However, using newer HIE passages and the MagMAX kit resulted in mean log fold increases for serially centrifuged GII.1, GII.4 and GII.6 (1.6, 2.3 and 1.8 log, respectively) that were similar to serially filtered samples. Therefore, HuNoV fecal sample processing and HIE age can affect virus replication in the HIE model. |
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| format | Article |
| id | doaj.art-9c978a04b1434fb7b7d11db189d8d628 |
| institution | Directory Open Access Journal |
| issn | 1999-4915 |
| language | English |
| last_indexed | 2024-03-07T22:11:15Z |
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| spelling | doaj.art-9c978a04b1434fb7b7d11db189d8d6282024-02-23T15:37:35ZengMDPI AGViruses1999-49152024-02-0116224110.3390/v16020241Replication of Human Norovirus in Human Intestinal Enteroids Is Affected by Fecal Sample ProcessingRevati Narwankar0Malak A. Esseili1Center for Food Safety, Department of Food Science and Technology, University of Georgia, Griffin, GA 30223, USACenter for Food Safety, Department of Food Science and Technology, University of Georgia, Griffin, GA 30223, USAHuman intestinal enteroids (HIEs) culture is an emerging model for assessing the infectivity of human noroviruses (HuNoVs). The model is based on detecting an increase in HuNoV RNA post-infection of HIEs. HuNoV fecal samples used for HIE infection are traditionally processed by serial filtration. Recently, processing HuNoV fecal samples by serial centrifugation was shown to retain vesicles containing HuNoV. The objective of this study was to investigate whether serially centrifuged fecal samples, RNA extraction kit (QIAamp versus MagMaX) and HIE age (newer versus older) affect HuNoV RNA fold increase in HIE. HuNoV GII.1, GII.4 and GII.6 fecal samples were prepared by serial centrifugation and filtration and the viral RNA in HIE was quantified at 1 and 72 h post-infection (hpi) following RNA extraction and RT-qPCR. The serially filtered GII.1, GII.4 and GII.6 showed successful replication in HIE, resulting in mean log increases of 2.2, 2 and 1.2, respectively, at 72 vs. 1 hpi. In contrast, only serially centrifuged GII.1 showed consistently successful replication. However, using newer HIE passages and the MagMAX kit resulted in mean log fold increases for serially centrifuged GII.1, GII.4 and GII.6 (1.6, 2.3 and 1.8 log, respectively) that were similar to serially filtered samples. Therefore, HuNoV fecal sample processing and HIE age can affect virus replication in the HIE model.https://www.mdpi.com/1999-4915/16/2/241human norovirushuman intestinal enteroidsreplication |
| spellingShingle | Revati Narwankar Malak A. Esseili Replication of Human Norovirus in Human Intestinal Enteroids Is Affected by Fecal Sample Processing Viruses human norovirus human intestinal enteroids replication |
| title | Replication of Human Norovirus in Human Intestinal Enteroids Is Affected by Fecal Sample Processing |
| title_full | Replication of Human Norovirus in Human Intestinal Enteroids Is Affected by Fecal Sample Processing |
| title_fullStr | Replication of Human Norovirus in Human Intestinal Enteroids Is Affected by Fecal Sample Processing |
| title_full_unstemmed | Replication of Human Norovirus in Human Intestinal Enteroids Is Affected by Fecal Sample Processing |
| title_short | Replication of Human Norovirus in Human Intestinal Enteroids Is Affected by Fecal Sample Processing |
| title_sort | replication of human norovirus in human intestinal enteroids is affected by fecal sample processing |
| topic | human norovirus human intestinal enteroids replication |
| url | https://www.mdpi.com/1999-4915/16/2/241 |
| work_keys_str_mv | AT revatinarwankar replicationofhumannorovirusinhumanintestinalenteroidsisaffectedbyfecalsampleprocessing AT malakaesseili replicationofhumannorovirusinhumanintestinalenteroidsisaffectedbyfecalsampleprocessing |