Rapid detection of H5 subtype avian influenza virus using CRISPR Cas13a based-lateral flow dipstick
Copyright © 2023 Li, Shang, Luo, Zhang, Meng, Feng, Jiang, Yu, Deng, Liu and Liu..
Due to its high mortality rate, highly pathogenic avian influenza (HPAI), a notifiable animal illness designated by the World Organisation for Animal Health (WOAH), has caused enormous financial losses to the poultry sector. The H5 subtype of avian influenza virus (H5-AIV) is regarded as the most common highly pathogenic avian influenza virus (HPAIV) that threatens public health and safety. Virus isolation and reverse transcription quantitative PCR (RT-qPCR) are usually used to detect H5-AIV and are important for the timely diagnosis and control of H5-AIV. However, these methods are time-consuming and require a significant amount of effort. In this study, we established a recombinase-aided amplification (RAA) combined with CRISPR-Cas13a and lateral flow dipstick (LFD) assay for the detection of H5-AIV. The results showed that the process can be completed within 40 min at 37°C. The method had a detection limit of 0.1 copy/μL, which was comparable to the RT-qPCR. There was no cross-reactivity with H3-AIV, H7-AIV, H9-AIV, H10-AIV, IBV, NDV, RVA and DAstV. The kappa value of RT-RAA-Cas13a-LFD and RT-qPCR in 380 clinical samples was 0.89 (κ>0.75). In conclusion, we established a convenient, efficient and accurate method to detect H5-AIV, and the results can be visualized and interpreted using LFD, which can be adapted to the needs of grassroots laboratories and field-deployable assays. This approach provides a new perspective for clinical H5-AIV diagnosis and has great potential for application in clinical quarantine of the poultry farming.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2023 |
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| Erschienen: |
2023 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:14 |
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| Enthalten in: |
Frontiers in microbiology - 14(2023) vom: 08., Seite 1283210 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Li, Yang [VerfasserIn] |
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| Links: |
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| Themen: |
CRISPR Cas13a |
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| Anmerkungen: |
Date Revised 14.12.2023 published: Electronic-eCollection Citation Status PubMed-not-MEDLINE |
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| doi: |
10.3389/fmicb.2023.1283210 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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| 520 | |a Copyright © 2023 Li, Shang, Luo, Zhang, Meng, Feng, Jiang, Yu, Deng, Liu and Liu. | ||
| 520 | |a Due to its high mortality rate, highly pathogenic avian influenza (HPAI), a notifiable animal illness designated by the World Organisation for Animal Health (WOAH), has caused enormous financial losses to the poultry sector. The H5 subtype of avian influenza virus (H5-AIV) is regarded as the most common highly pathogenic avian influenza virus (HPAIV) that threatens public health and safety. Virus isolation and reverse transcription quantitative PCR (RT-qPCR) are usually used to detect H5-AIV and are important for the timely diagnosis and control of H5-AIV. However, these methods are time-consuming and require a significant amount of effort. In this study, we established a recombinase-aided amplification (RAA) combined with CRISPR-Cas13a and lateral flow dipstick (LFD) assay for the detection of H5-AIV. The results showed that the process can be completed within 40 min at 37°C. The method had a detection limit of 0.1 copy/μL, which was comparable to the RT-qPCR. There was no cross-reactivity with H3-AIV, H7-AIV, H9-AIV, H10-AIV, IBV, NDV, RVA and DAstV. The kappa value of RT-RAA-Cas13a-LFD and RT-qPCR in 380 clinical samples was 0.89 (κ>0.75). In conclusion, we established a convenient, efficient and accurate method to detect H5-AIV, and the results can be visualized and interpreted using LFD, which can be adapted to the needs of grassroots laboratories and field-deployable assays. This approach provides a new perspective for clinical H5-AIV diagnosis and has great potential for application in clinical quarantine of the poultry farming | ||
| 650 | 4 | |a Journal Article | |
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| 650 | 4 | |a H5-AIV | |
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| 650 | 4 | |a lateral flow dipstick | |
| 650 | 4 | |a rapid diagnosis | |
| 700 | 1 | |a Shang, Jiajing |e verfasserin |4 aut | |
| 700 | 1 | |a Luo, Juan |e verfasserin |4 aut | |
| 700 | 1 | |a Zhang, Fuyou |e verfasserin |4 aut | |
| 700 | 1 | |a Meng, Ge |e verfasserin |4 aut | |
| 700 | 1 | |a Feng, Yingjie |e verfasserin |4 aut | |
| 700 | 1 | |a Jiang, Wenming |e verfasserin |4 aut | |
| 700 | 1 | |a Yu, Xiaohui |e verfasserin |4 aut | |
| 700 | 1 | |a Deng, Chunran |e verfasserin |4 aut | |
| 700 | 1 | |a Liu, Guanhui |e verfasserin |4 aut | |
| 700 | 1 | |a Liu, Hualei |e verfasserin |4 aut | |
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