Rapid molecular detection of Senecavirus A based on reverse transcription loop-mediated isothermal amplification (RT-LAMP) and CRISPR/Cas12a
IntroductionSenecavirus A (SVA), an emerging vesicular pathogen, is responsible for porcine idiopathic vesicular disease (PIVD). This disease is closely associated with porcine vesicular disease and acute neonatal piglet mortality, presenting a substantial threat to the global swine industry. At pre...
Saved in:
| Main Authors: | , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Frontiers Media S.A.
2025-04-01
|
| Series: | Frontiers in Bioengineering and Biotechnology |
| Subjects: | |
| Online Access: | https://www.frontiersin.org/articles/10.3389/fbioe.2025.1451125/full |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1849735741499768832 |
|---|---|
| author | Chenghui Jiang Chenghui Jiang Huibao Wang Rongxia Guo Rui Yang Xiaoming Li Ping Liu Jing Wang Jincai Yang Yanyan Chang Yanyan Chang Qiaoying Zeng |
| author_facet | Chenghui Jiang Chenghui Jiang Huibao Wang Rongxia Guo Rui Yang Xiaoming Li Ping Liu Jing Wang Jincai Yang Yanyan Chang Yanyan Chang Qiaoying Zeng |
| author_sort | Chenghui Jiang |
| collection | DOAJ |
| description | IntroductionSenecavirus A (SVA), an emerging vesicular pathogen, is responsible for porcine idiopathic vesicular disease (PIVD). This disease is closely associated with porcine vesicular disease and acute neonatal piglet mortality, presenting a substantial threat to the global swine industry. At present, the absence of effective drugs or vaccines for treating the disease makes accurate diagnosis of SVA of paramount importance for the effective prevention and control of the disease.MethodsIn this study, we combined reverse transcription loop-mediated isothermal amplification (RT-LAMP) and Clustered Regularly Interspaced Short Palindromic Repeats and CRISPR-associated protein12a (CRISPR/Cas12a) using a dual-labelled fluorescence quencher or fluorescent biotin single-stranded DNA reporter molecule to develop two rapid, reliable, and portable visual SVA assays: RT-LAMP-Cas12a-FQ and RT-LAMP-Cas12a-FB.ResultsThe two methods exhibited comparable detection limits, with 9.6 copies/μL achieved in 40 and 45 minutes, respectively. They did not cross-react with non-target nucleic acids extracted from other related viruses and showed high specificity for SVA RNA detection. Furthermore, the methods demonstrated satisfactory performance in detecting 69 porcine adventitious samples, with no significant differences from that of quantitative reverse transcription polymerase chain reaction (RT-qPCR).DiscussionIn summary, the RT-LAMP-Cas12a-FQ and RT-LAMP-Cas12a-FB methods developed are promising for early detection and routine surveillance of porcine SVA in resource-poor areas. |
| format | Article |
| id | doaj-art-4f5b54d2a9a044d1b702f5fb93a2a69c |
| institution | DOAJ |
| issn | 2296-4185 |
| language | English |
| publishDate | 2025-04-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Bioengineering and Biotechnology |
| spelling | doaj-art-4f5b54d2a9a044d1b702f5fb93a2a69c2025-08-20T03:07:28ZengFrontiers Media S.A.Frontiers in Bioengineering and Biotechnology2296-41852025-04-011310.3389/fbioe.2025.14511251451125Rapid molecular detection of Senecavirus A based on reverse transcription loop-mediated isothermal amplification (RT-LAMP) and CRISPR/Cas12aChenghui Jiang0Chenghui Jiang1Huibao Wang2Rongxia Guo3Rui Yang4Xiaoming Li5Ping Liu6Jing Wang7Jincai Yang8Yanyan Chang9Yanyan Chang10Qiaoying Zeng11College of Veterinary Medicine, Gansu Agricultural University, Lanzhou, Gansu, ChinaChina Agricultural Veterinary Biological Science and Technology Co., Ltd., Lanzhou, Gansu, ChinaCollege of Environmental Engineering, Gansu Forestry Voctech University, Tianshui, Gansu, ChinaCollege of Veterinary Medicine, Gansu Agricultural University, Lanzhou, Gansu, ChinaChina Agricultural Veterinary Biological Science and Technology Co., Ltd., Lanzhou, Gansu, ChinaChina Agricultural Veterinary Biological Science and Technology Co., Ltd., Lanzhou, Gansu, ChinaChina Agricultural Veterinary Biological Science and Technology Co., Ltd., Lanzhou, Gansu, ChinaChina Agricultural Veterinary Biological Science and Technology Co., Ltd., Lanzhou, Gansu, ChinaChina Agricultural Veterinary Biological Science and Technology Co., Ltd., Lanzhou, Gansu, ChinaCollege of Veterinary Medicine, Gansu Agricultural University, Lanzhou, Gansu, ChinaChina Agricultural Veterinary Biological Science and Technology Co., Ltd., Lanzhou, Gansu, ChinaCollege of Veterinary Medicine, Gansu Agricultural University, Lanzhou, Gansu, ChinaIntroductionSenecavirus A (SVA), an emerging vesicular pathogen, is responsible for porcine idiopathic vesicular disease (PIVD). This disease is closely associated with porcine vesicular disease and acute neonatal piglet mortality, presenting a substantial threat to the global swine industry. At present, the absence of effective drugs or vaccines for treating the disease makes accurate diagnosis of SVA of paramount importance for the effective prevention and control of the disease.MethodsIn this study, we combined reverse transcription loop-mediated isothermal amplification (RT-LAMP) and Clustered Regularly Interspaced Short Palindromic Repeats and CRISPR-associated protein12a (CRISPR/Cas12a) using a dual-labelled fluorescence quencher or fluorescent biotin single-stranded DNA reporter molecule to develop two rapid, reliable, and portable visual SVA assays: RT-LAMP-Cas12a-FQ and RT-LAMP-Cas12a-FB.ResultsThe two methods exhibited comparable detection limits, with 9.6 copies/μL achieved in 40 and 45 minutes, respectively. They did not cross-react with non-target nucleic acids extracted from other related viruses and showed high specificity for SVA RNA detection. Furthermore, the methods demonstrated satisfactory performance in detecting 69 porcine adventitious samples, with no significant differences from that of quantitative reverse transcription polymerase chain reaction (RT-qPCR).DiscussionIn summary, the RT-LAMP-Cas12a-FQ and RT-LAMP-Cas12a-FB methods developed are promising for early detection and routine surveillance of porcine SVA in resource-poor areas.https://www.frontiersin.org/articles/10.3389/fbioe.2025.1451125/fullSenecavirus ART-LAMPCRISPR/Cas12anucleic acid detectionvisualisation |
| spellingShingle | Chenghui Jiang Chenghui Jiang Huibao Wang Rongxia Guo Rui Yang Xiaoming Li Ping Liu Jing Wang Jincai Yang Yanyan Chang Yanyan Chang Qiaoying Zeng Rapid molecular detection of Senecavirus A based on reverse transcription loop-mediated isothermal amplification (RT-LAMP) and CRISPR/Cas12a Frontiers in Bioengineering and Biotechnology Senecavirus A RT-LAMP CRISPR/Cas12a nucleic acid detection visualisation |
| title | Rapid molecular detection of Senecavirus A based on reverse transcription loop-mediated isothermal amplification (RT-LAMP) and CRISPR/Cas12a |
| title_full | Rapid molecular detection of Senecavirus A based on reverse transcription loop-mediated isothermal amplification (RT-LAMP) and CRISPR/Cas12a |
| title_fullStr | Rapid molecular detection of Senecavirus A based on reverse transcription loop-mediated isothermal amplification (RT-LAMP) and CRISPR/Cas12a |
| title_full_unstemmed | Rapid molecular detection of Senecavirus A based on reverse transcription loop-mediated isothermal amplification (RT-LAMP) and CRISPR/Cas12a |
| title_short | Rapid molecular detection of Senecavirus A based on reverse transcription loop-mediated isothermal amplification (RT-LAMP) and CRISPR/Cas12a |
| title_sort | rapid molecular detection of senecavirus a based on reverse transcription loop mediated isothermal amplification rt lamp and crispr cas12a |
| topic | Senecavirus A RT-LAMP CRISPR/Cas12a nucleic acid detection visualisation |
| url | https://www.frontiersin.org/articles/10.3389/fbioe.2025.1451125/full |
| work_keys_str_mv | AT chenghuijiang rapidmoleculardetectionofsenecavirusabasedonreversetranscriptionloopmediatedisothermalamplificationrtlampandcrisprcas12a AT chenghuijiang rapidmoleculardetectionofsenecavirusabasedonreversetranscriptionloopmediatedisothermalamplificationrtlampandcrisprcas12a AT huibaowang rapidmoleculardetectionofsenecavirusabasedonreversetranscriptionloopmediatedisothermalamplificationrtlampandcrisprcas12a AT rongxiaguo rapidmoleculardetectionofsenecavirusabasedonreversetranscriptionloopmediatedisothermalamplificationrtlampandcrisprcas12a AT ruiyang rapidmoleculardetectionofsenecavirusabasedonreversetranscriptionloopmediatedisothermalamplificationrtlampandcrisprcas12a AT xiaomingli rapidmoleculardetectionofsenecavirusabasedonreversetranscriptionloopmediatedisothermalamplificationrtlampandcrisprcas12a AT pingliu rapidmoleculardetectionofsenecavirusabasedonreversetranscriptionloopmediatedisothermalamplificationrtlampandcrisprcas12a AT jingwang rapidmoleculardetectionofsenecavirusabasedonreversetranscriptionloopmediatedisothermalamplificationrtlampandcrisprcas12a AT jincaiyang rapidmoleculardetectionofsenecavirusabasedonreversetranscriptionloopmediatedisothermalamplificationrtlampandcrisprcas12a AT yanyanchang rapidmoleculardetectionofsenecavirusabasedonreversetranscriptionloopmediatedisothermalamplificationrtlampandcrisprcas12a AT yanyanchang rapidmoleculardetectionofsenecavirusabasedonreversetranscriptionloopmediatedisothermalamplificationrtlampandcrisprcas12a AT qiaoyingzeng rapidmoleculardetectionofsenecavirusabasedonreversetranscriptionloopmediatedisothermalamplificationrtlampandcrisprcas12a |