A Novel Sandwich ELASA Based on Aptamer for Detection of Largemouth Bass Virus (LMBV)
Largemouth bass virus (LMBV) is a major viral pathogen in largemouth bass culture, usually causing high mortality and heavy economic losses. Accurate and early detection of LMBV is crucial for diagnosis and control of the diseases caused by LMBV. Previously, we selected the specific aptamers, LA38 a...
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| Format: | Article |
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MDPI AG
2022-04-01
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| Series: | Viruses |
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| Online Access: | https://www.mdpi.com/1999-4915/14/5/945 |
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| author | Xinyue Zhang Zemiao Zhang Junrong Li Xiaohong Huang Jingguang Wei Jiahui Yang Lingfeng Guan Xiaozhi Wen Shaowen Wang Qiwei Qin |
| author_facet | Xinyue Zhang Zemiao Zhang Junrong Li Xiaohong Huang Jingguang Wei Jiahui Yang Lingfeng Guan Xiaozhi Wen Shaowen Wang Qiwei Qin |
| author_sort | Xinyue Zhang |
| collection | DOAJ |
| description | Largemouth bass virus (LMBV) is a major viral pathogen in largemouth bass culture, usually causing high mortality and heavy economic losses. Accurate and early detection of LMBV is crucial for diagnosis and control of the diseases caused by LMBV. Previously, we selected the specific aptamers, LA38 and LA13, targeting LMBV by systematic evolution of ligands by exponential enrichment (SELEX). In this study, we further generated truncated LA38 and LA13 (named as LA38s and LA13s) with high specificity and affinities and developed an aptamer-based sandwich enzyme-linked apta-sorbent assay (ELASA) for LMBV diagnosis. The sandwich ELASA showed high specificity and sensitivity for the LMBV detection, without cross reaction with other viruses. The detection limit of the ELASA was as low as 1.25 × 10<sup>2</sup> LMBV-infected cells, and the incubation time of the lysate and biotin labeled aptamer was as short as 10 min. The ELASA could still detect LMBV infection in spleen lysates at dilutions of 1/25, with good consistency of qRT-PCR. For the fish samples collected from the field, the sensitivity of ELASA was 13.3% less than PCR, but the ELASA was much more convenient and less time consuming. The procedure of ELASA mainly requires washing and incubation, with completion in approximately 4 h. The sandwich ELASA offers a useful tool to rapidly detect LMBV rapidly, contributing to control and prevention of LMBV infection. |
| first_indexed | 2024-03-10T01:37:18Z |
| format | Article |
| id | doaj.art-4413c6c876074d73a826dcb7e6392cb1 |
| institution | Directory Open Access Journal |
| issn | 1999-4915 |
| language | English |
| last_indexed | 2024-03-10T01:37:18Z |
| publishDate | 2022-04-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Viruses |
| spelling | doaj.art-4413c6c876074d73a826dcb7e6392cb12023-11-23T13:30:42ZengMDPI AGViruses1999-49152022-04-0114594510.3390/v14050945A Novel Sandwich ELASA Based on Aptamer for Detection of Largemouth Bass Virus (LMBV)Xinyue Zhang0Zemiao Zhang1Junrong Li2Xiaohong Huang3Jingguang Wei4Jiahui Yang5Lingfeng Guan6Xiaozhi Wen7Shaowen Wang8Qiwei Qin9College of Marine Sciences, South China Agricultural University, Guangdong Laboratory for Lingnan Modern Agriculture, Guangzhou 510642, ChinaCollege of Marine Sciences, South China Agricultural University, Guangdong Laboratory for Lingnan Modern Agriculture, Guangzhou 510642, ChinaCollege of Marine Sciences, South China Agricultural University, Guangdong Laboratory for Lingnan Modern Agriculture, Guangzhou 510642, ChinaCollege of Marine Sciences, South China Agricultural University, Guangdong Laboratory for Lingnan Modern Agriculture, Guangzhou 510642, ChinaCollege of Marine Sciences, South China Agricultural University, Guangdong Laboratory for Lingnan Modern Agriculture, Guangzhou 510642, ChinaCollege of Marine Sciences, South China Agricultural University, Guangdong Laboratory for Lingnan Modern Agriculture, Guangzhou 510642, ChinaCollege of Marine Sciences, South China Agricultural University, Guangdong Laboratory for Lingnan Modern Agriculture, Guangzhou 510642, ChinaCollege of Marine Sciences, South China Agricultural University, Guangdong Laboratory for Lingnan Modern Agriculture, Guangzhou 510642, ChinaCollege of Marine Sciences, South China Agricultural University, Guangdong Laboratory for Lingnan Modern Agriculture, Guangzhou 510642, ChinaCollege of Marine Sciences, South China Agricultural University, Guangdong Laboratory for Lingnan Modern Agriculture, Guangzhou 510642, ChinaLargemouth bass virus (LMBV) is a major viral pathogen in largemouth bass culture, usually causing high mortality and heavy economic losses. Accurate and early detection of LMBV is crucial for diagnosis and control of the diseases caused by LMBV. Previously, we selected the specific aptamers, LA38 and LA13, targeting LMBV by systematic evolution of ligands by exponential enrichment (SELEX). In this study, we further generated truncated LA38 and LA13 (named as LA38s and LA13s) with high specificity and affinities and developed an aptamer-based sandwich enzyme-linked apta-sorbent assay (ELASA) for LMBV diagnosis. The sandwich ELASA showed high specificity and sensitivity for the LMBV detection, without cross reaction with other viruses. The detection limit of the ELASA was as low as 1.25 × 10<sup>2</sup> LMBV-infected cells, and the incubation time of the lysate and biotin labeled aptamer was as short as 10 min. The ELASA could still detect LMBV infection in spleen lysates at dilutions of 1/25, with good consistency of qRT-PCR. For the fish samples collected from the field, the sensitivity of ELASA was 13.3% less than PCR, but the ELASA was much more convenient and less time consuming. The procedure of ELASA mainly requires washing and incubation, with completion in approximately 4 h. The sandwich ELASA offers a useful tool to rapidly detect LMBV rapidly, contributing to control and prevention of LMBV infection.https://www.mdpi.com/1999-4915/14/5/945ELASAaptamerlargemouth bass virusrapid detection |
| spellingShingle | Xinyue Zhang Zemiao Zhang Junrong Li Xiaohong Huang Jingguang Wei Jiahui Yang Lingfeng Guan Xiaozhi Wen Shaowen Wang Qiwei Qin A Novel Sandwich ELASA Based on Aptamer for Detection of Largemouth Bass Virus (LMBV) Viruses ELASA aptamer largemouth bass virus rapid detection |
| title | A Novel Sandwich ELASA Based on Aptamer for Detection of Largemouth Bass Virus (LMBV) |
| title_full | A Novel Sandwich ELASA Based on Aptamer for Detection of Largemouth Bass Virus (LMBV) |
| title_fullStr | A Novel Sandwich ELASA Based on Aptamer for Detection of Largemouth Bass Virus (LMBV) |
| title_full_unstemmed | A Novel Sandwich ELASA Based on Aptamer for Detection of Largemouth Bass Virus (LMBV) |
| title_short | A Novel Sandwich ELASA Based on Aptamer for Detection of Largemouth Bass Virus (LMBV) |
| title_sort | novel sandwich elasa based on aptamer for detection of largemouth bass virus lmbv |
| topic | ELASA aptamer largemouth bass virus rapid detection |
| url | https://www.mdpi.com/1999-4915/14/5/945 |
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