Rapid and Visual Detection of Actinidia Chlorotic Ringspot-Associated Virus Using One-Step Reverse-Transcription Recombinase Polymerase Amplification Combined with Lateral Flow Dipstick Assay
Actinidia chlorotic ringspot-associated virus (AcCRaV) occurs widely in major kiwifruit producing areas of China and is often accompanied by coinfecting viruses, affecting the growth, yield, and quality of kiwifruit. Therefore, a rapid and sensitive detection method is crucial for diagnosing and developing effective AcCRaV management strategies. In this study, a one-step reverse-transcription recombinase polymerase amplification combined with a lateral flow dipstick (RT-RPA-LFD) assay was developed for rapid detection of AcCRaV. Specific primers and a probe were designed based on the conserved region of the coat protein gene sequence of AcCRaV. The one-step RT-RPA reaction can be performed at 35 and 40°C within 10 to 30 min, and the amplification results can be read directly on the LFD within 5 min. The detection limit of the one-step RT-RPA-LFD assay was 10-8 ng (about 20 viral copies), which was equal with one-step RT-qPCR and 100 times more sensitive than one-step RT-PCR. Moreover, the one-step RT-RPA-LFD assay was successfully applied to detect AcCRaV from crude extracts, and the entire detection process can be completed within 40 min. These results indicate that the RT-RPA-LFD assay is a simple, rapid, and sensitive strategy that can be used for accurate diagnosis of AcCRaV-infected kiwifruit plants in the field. To our knowledge, this is the first study applying the one-step RT-RPA-LFD assay to detect a kiwifruit virus.
| Medienart: |
E-Artikel |
|---|
| Erscheinungsjahr: |
2023 |
|---|---|
| Erschienen: |
2023 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:107 |
|---|---|
| Enthalten in: |
Plant disease - 107(2023), 12 vom: 01. Dez., Seite 3701-3707 |
| Sprache: |
Englisch |
|---|
| Beteiligte Personen: |
Zhang, A-Ling [VerfasserIn] |
|---|
| Links: |
|---|
| Themen: |
AcCRaV |
|---|
| Anmerkungen: |
Date Completed 01.01.2024 Date Revised 01.01.2024 published: Print-Electronic Citation Status MEDLINE |
|---|
| doi: |
10.1094/PDIS-02-23-0270-SR |
|---|
| funding: |
|
|---|---|
| Förderinstitution / Projekttitel: |
|
| PPN (Katalog-ID): |
NLM359675883 |
|---|
| LEADER | 01000caa a22002652 4500 | ||
|---|---|---|---|
| 001 | NLM359675883 | ||
| 003 | DE-627 | ||
| 005 | 20240108140724.0 | ||
| 007 | cr uuu---uuuuu | ||
| 008 | 231226s2023 xx |||||o 00| ||eng c | ||
| 024 | 7 | |a 10.1094/PDIS-02-23-0270-SR |2 doi | |
| 028 | 5 | 2 | |a pubmed24n1246.xml |
| 035 | |a (DE-627)NLM359675883 | ||
| 035 | |a (NLM)37467124 | ||
| 040 | |a DE-627 |b ger |c DE-627 |e rakwb | ||
| 041 | |a eng | ||
| 100 | 1 | |a Zhang, A-Ling |e verfasserin |4 aut | |
| 245 | 1 | 0 | |a Rapid and Visual Detection of Actinidia Chlorotic Ringspot-Associated Virus Using One-Step Reverse-Transcription Recombinase Polymerase Amplification Combined with Lateral Flow Dipstick Assay |
| 264 | 1 | |c 2023 | |
| 336 | |a Text |b txt |2 rdacontent | ||
| 337 | |a ƒaComputermedien |b c |2 rdamedia | ||
| 338 | |a ƒa Online-Ressource |b cr |2 rdacarrier | ||
| 500 | |a Date Completed 01.01.2024 | ||
| 500 | |a Date Revised 01.01.2024 | ||
| 500 | |a published: Print-Electronic | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a Actinidia chlorotic ringspot-associated virus (AcCRaV) occurs widely in major kiwifruit producing areas of China and is often accompanied by coinfecting viruses, affecting the growth, yield, and quality of kiwifruit. Therefore, a rapid and sensitive detection method is crucial for diagnosing and developing effective AcCRaV management strategies. In this study, a one-step reverse-transcription recombinase polymerase amplification combined with a lateral flow dipstick (RT-RPA-LFD) assay was developed for rapid detection of AcCRaV. Specific primers and a probe were designed based on the conserved region of the coat protein gene sequence of AcCRaV. The one-step RT-RPA reaction can be performed at 35 and 40°C within 10 to 30 min, and the amplification results can be read directly on the LFD within 5 min. The detection limit of the one-step RT-RPA-LFD assay was 10-8 ng (about 20 viral copies), which was equal with one-step RT-qPCR and 100 times more sensitive than one-step RT-PCR. Moreover, the one-step RT-RPA-LFD assay was successfully applied to detect AcCRaV from crude extracts, and the entire detection process can be completed within 40 min. These results indicate that the RT-RPA-LFD assay is a simple, rapid, and sensitive strategy that can be used for accurate diagnosis of AcCRaV-infected kiwifruit plants in the field. To our knowledge, this is the first study applying the one-step RT-RPA-LFD assay to detect a kiwifruit virus | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a AcCRaV | |
| 650 | 4 | |a RT-RPA | |
| 650 | 4 | |a lateral flow dipstick | |
| 650 | 4 | |a on-site application | |
| 650 | 4 | |a rapid diagnosis | |
| 650 | 7 | |a Recombinases |2 NLM | |
| 700 | 1 | |a Shi, Xia |e verfasserin |4 aut | |
| 700 | 1 | |a Xie, Cuijuan |e verfasserin |4 aut | |
| 700 | 1 | |a Yu, Feng |e verfasserin |4 aut | |
| 700 | 1 | |a Gao, Zhixiong |e verfasserin |4 aut | |
| 700 | 1 | |a Xu, Yan |e verfasserin |4 aut | |
| 700 | 1 | |a Liu, Zhande |e verfasserin |4 aut | |
| 773 | 0 | 8 | |i Enthalten in |t Plant disease |d 1997 |g 107(2023), 12 vom: 01. Dez., Seite 3701-3707 |w (DE-627)NLM098181742 |x 0191-2917 |7 nnns |
| 773 | 1 | 8 | |g volume:107 |g year:2023 |g number:12 |g day:01 |g month:12 |g pages:3701-3707 |
| 856 | 4 | 0 | |u http://dx.doi.org/10.1094/PDIS-02-23-0270-SR |3 Volltext |
| 912 | |a GBV_USEFLAG_A | ||
| 912 | |a GBV_NLM | ||
| 951 | |a AR | ||
| 952 | |d 107 |j 2023 |e 12 |b 01 |c 12 |h 3701-3707 | ||