Cargando…
CRISPR-Cas13a-based detection method for avian influenza virus
Avian influenza virus (AIV) causes huge losses to the global poultry industry and poses a threat to humans and other mammals. Fast, sensitive, and portable diagnostic methods are essential for efficient avian influenza control. Here, a clustered regularly interspaced short palindromic repeats (CRISP...
Autores principales: | , , , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Frontiers Media S.A.
2023
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10598603/ https://www.ncbi.nlm.nih.gov/pubmed/37886067 http://dx.doi.org/10.3389/fmicb.2023.1288951 |
_version_ | 1785125591303847936 |
---|---|
author | Wu, Yuhan Zhan, Jiaxing Shan, Zhaomeng Li, Yanbing Liu, Yining Li, Yan Wang, Yixin Liu, Zhe Wen, Xuexia Wang, Xiurong |
author_facet | Wu, Yuhan Zhan, Jiaxing Shan, Zhaomeng Li, Yanbing Liu, Yining Li, Yan Wang, Yixin Liu, Zhe Wen, Xuexia Wang, Xiurong |
author_sort | Wu, Yuhan |
collection | PubMed |
description | Avian influenza virus (AIV) causes huge losses to the global poultry industry and poses a threat to humans and other mammals. Fast, sensitive, and portable diagnostic methods are essential for efficient avian influenza control. Here, a clustered regularly interspaced short palindromic repeats (CRISPR)-Cas13a based platform was developed to detect AIV. This novel method was developed to specifically detect H1–H16 subtypes of AIV with fluorescence and lateral flow-based readouts and exhibited no cross-reactivity with Newcastle disease virus, avian infectious bronchitis virus, or infectious bursal disease virus. The limit of detection was determined to be 69 and 690 copies/μL using fluorescence and lateral flow as readouts, respectively. The developed assay exhibited 100% consistency with quantitative real-time polymerase chain reaction in detecting clinical samples. The heating of unextracted diagnostic samples to obliterate nuclease treatment was introduced to detect viral RNA without nucleic acid extraction. Single-step optimization was used to perform reverse transcription, recombinase polymerase amplification, and CRISPR-Cas13a detection in a tube. These advances resulted in an optimized assay that could specifically detect AIV with simplified procedures and reduced contamination risk, highlighting the potential to be used in point-of-care testing. |
format | Online Article Text |
id | pubmed-10598603 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-105986032023-10-26 CRISPR-Cas13a-based detection method for avian influenza virus Wu, Yuhan Zhan, Jiaxing Shan, Zhaomeng Li, Yanbing Liu, Yining Li, Yan Wang, Yixin Liu, Zhe Wen, Xuexia Wang, Xiurong Front Microbiol Microbiology Avian influenza virus (AIV) causes huge losses to the global poultry industry and poses a threat to humans and other mammals. Fast, sensitive, and portable diagnostic methods are essential for efficient avian influenza control. Here, a clustered regularly interspaced short palindromic repeats (CRISPR)-Cas13a based platform was developed to detect AIV. This novel method was developed to specifically detect H1–H16 subtypes of AIV with fluorescence and lateral flow-based readouts and exhibited no cross-reactivity with Newcastle disease virus, avian infectious bronchitis virus, or infectious bursal disease virus. The limit of detection was determined to be 69 and 690 copies/μL using fluorescence and lateral flow as readouts, respectively. The developed assay exhibited 100% consistency with quantitative real-time polymerase chain reaction in detecting clinical samples. The heating of unextracted diagnostic samples to obliterate nuclease treatment was introduced to detect viral RNA without nucleic acid extraction. Single-step optimization was used to perform reverse transcription, recombinase polymerase amplification, and CRISPR-Cas13a detection in a tube. These advances resulted in an optimized assay that could specifically detect AIV with simplified procedures and reduced contamination risk, highlighting the potential to be used in point-of-care testing. Frontiers Media S.A. 2023-10-11 /pmc/articles/PMC10598603/ /pubmed/37886067 http://dx.doi.org/10.3389/fmicb.2023.1288951 Text en Copyright © 2023 Wu, Zhan, Shan, Li, Liu, Li, Wang, Liu, Wen and Wang. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
spellingShingle | Microbiology Wu, Yuhan Zhan, Jiaxing Shan, Zhaomeng Li, Yanbing Liu, Yining Li, Yan Wang, Yixin Liu, Zhe Wen, Xuexia Wang, Xiurong CRISPR-Cas13a-based detection method for avian influenza virus |
title | CRISPR-Cas13a-based detection method for avian influenza virus |
title_full | CRISPR-Cas13a-based detection method for avian influenza virus |
title_fullStr | CRISPR-Cas13a-based detection method for avian influenza virus |
title_full_unstemmed | CRISPR-Cas13a-based detection method for avian influenza virus |
title_short | CRISPR-Cas13a-based detection method for avian influenza virus |
title_sort | crispr-cas13a-based detection method for avian influenza virus |
topic | Microbiology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10598603/ https://www.ncbi.nlm.nih.gov/pubmed/37886067 http://dx.doi.org/10.3389/fmicb.2023.1288951 |
work_keys_str_mv | AT wuyuhan crisprcas13abaseddetectionmethodforavianinfluenzavirus AT zhanjiaxing crisprcas13abaseddetectionmethodforavianinfluenzavirus AT shanzhaomeng crisprcas13abaseddetectionmethodforavianinfluenzavirus AT liyanbing crisprcas13abaseddetectionmethodforavianinfluenzavirus AT liuyining crisprcas13abaseddetectionmethodforavianinfluenzavirus AT liyan crisprcas13abaseddetectionmethodforavianinfluenzavirus AT wangyixin crisprcas13abaseddetectionmethodforavianinfluenzavirus AT liuzhe crisprcas13abaseddetectionmethodforavianinfluenzavirus AT wenxuexia crisprcas13abaseddetectionmethodforavianinfluenzavirus AT wangxiurong crisprcas13abaseddetectionmethodforavianinfluenzavirus |