Monoclonal antibody-based colloid gold immunochromatographic strip for the rapid detection of Tomato zonate spot tospovirus
Abstract Background Tomato zonate spot virus (TZSV), a new species of genus Tospovirus, caused significant losses in yield and problems in quality of many important vegetables and ornamentals in Southwest China and posed a serious threat to important economic crops for the local farmers. A convenien...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2018-01-01
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| Series: | Virology Journal |
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| Online Access: | http://link.springer.com/article/10.1186/s12985-018-0919-5 |
| _version_ | 1811231302104383488 |
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| author | Yanbing Niu Defu Wang Liyan Cui Baoxia Wang Xiaojing Pang Peixia Yu |
| author_facet | Yanbing Niu Defu Wang Liyan Cui Baoxia Wang Xiaojing Pang Peixia Yu |
| author_sort | Yanbing Niu |
| collection | DOAJ |
| description | Abstract Background Tomato zonate spot virus (TZSV), a new species of genus Tospovirus, caused significant losses in yield and problems in quality of many important vegetables and ornamentals in Southwest China and posed a serious threat to important economic crops for the local farmers. A convenient and reliable method was urgently needed for rapid detection and surveillance of TZSV. Methods The nucleocapsid protein (N) of TZSV was expressed in Escherichia coli and purified, and was used as the antigen to immunize BALB/c mice. Three monoclonal antibodies (mAbs) 3A2, 5D2 and 5F7 against TZSV were obtained through the hybridoma technique. The mAb 3A2 was conjugated with colloid gold as detecting reagent; mAb 5D2 was coated on a porous nitrocellulose membrane as the detection line and protein A was coated as the control line respectively. The colloid gold immunochromatographic (GICA) strip was assembled. Results The analysis of Dot-ELISA and Western blot showed that the obtained three independent lines of mAbs 3A2, 5D2 and 5F7 specifically recognized TZSV N. Based on the assembly of GICA strip, the detection of TZSV was achieved by loading the infected sap onto the test strip for visual inspection. The analysis could be completed within 5–10 min. No cross-reaction occurred between TZSV and other tested viruses. The visual detection limit of the test strip for TZSV was 800 fold dilutions of TZSV-infected leaf samples. Conclusion The mAbs were specific and the colloidal GICA strip developed in this study was convenient, fast and reliable for the detection of TZSV. The method could be applied for the rapid diagnosis and surveillance of TZSV in the field. |
| first_indexed | 2024-04-12T10:42:29Z |
| format | Article |
| id | doaj.art-5152fe98c32843c588e97052584c848f |
| institution | Directory Open Access Journal |
| issn | 1743-422X |
| language | English |
| last_indexed | 2024-04-12T10:42:29Z |
| publishDate | 2018-01-01 |
| publisher | BMC |
| record_format | Article |
| series | Virology Journal |
| spelling | doaj.art-5152fe98c32843c588e97052584c848f2022-12-22T03:36:33ZengBMCVirology Journal1743-422X2018-01-011511910.1186/s12985-018-0919-5Monoclonal antibody-based colloid gold immunochromatographic strip for the rapid detection of Tomato zonate spot tospovirusYanbing Niu0Defu Wang1Liyan Cui2Baoxia Wang3Xiaojing Pang4Peixia Yu5College of Life Sciences, Shanxi Agricultural UniversityCollege of Life Sciences, Shanxi Agricultural UniversityCollege of Life Sciences, Shanxi Agricultural UniversityCollege of Life Sciences, Shanxi Agricultural UniversityCollege of Life Sciences, Shanxi Agricultural UniversityCollege of Life Sciences, Shanxi Agricultural UniversityAbstract Background Tomato zonate spot virus (TZSV), a new species of genus Tospovirus, caused significant losses in yield and problems in quality of many important vegetables and ornamentals in Southwest China and posed a serious threat to important economic crops for the local farmers. A convenient and reliable method was urgently needed for rapid detection and surveillance of TZSV. Methods The nucleocapsid protein (N) of TZSV was expressed in Escherichia coli and purified, and was used as the antigen to immunize BALB/c mice. Three monoclonal antibodies (mAbs) 3A2, 5D2 and 5F7 against TZSV were obtained through the hybridoma technique. The mAb 3A2 was conjugated with colloid gold as detecting reagent; mAb 5D2 was coated on a porous nitrocellulose membrane as the detection line and protein A was coated as the control line respectively. The colloid gold immunochromatographic (GICA) strip was assembled. Results The analysis of Dot-ELISA and Western blot showed that the obtained three independent lines of mAbs 3A2, 5D2 and 5F7 specifically recognized TZSV N. Based on the assembly of GICA strip, the detection of TZSV was achieved by loading the infected sap onto the test strip for visual inspection. The analysis could be completed within 5–10 min. No cross-reaction occurred between TZSV and other tested viruses. The visual detection limit of the test strip for TZSV was 800 fold dilutions of TZSV-infected leaf samples. Conclusion The mAbs were specific and the colloidal GICA strip developed in this study was convenient, fast and reliable for the detection of TZSV. The method could be applied for the rapid diagnosis and surveillance of TZSV in the field.http://link.springer.com/article/10.1186/s12985-018-0919-5Tomato zonate spot virus (TZSV)Monoclonal antibodiesColloid gold immunochromatographic (GICA) stripRapid detection |
| spellingShingle | Yanbing Niu Defu Wang Liyan Cui Baoxia Wang Xiaojing Pang Peixia Yu Monoclonal antibody-based colloid gold immunochromatographic strip for the rapid detection of Tomato zonate spot tospovirus Virology Journal Tomato zonate spot virus (TZSV) Monoclonal antibodies Colloid gold immunochromatographic (GICA) strip Rapid detection |
| title | Monoclonal antibody-based colloid gold immunochromatographic strip for the rapid detection of Tomato zonate spot tospovirus |
| title_full | Monoclonal antibody-based colloid gold immunochromatographic strip for the rapid detection of Tomato zonate spot tospovirus |
| title_fullStr | Monoclonal antibody-based colloid gold immunochromatographic strip for the rapid detection of Tomato zonate spot tospovirus |
| title_full_unstemmed | Monoclonal antibody-based colloid gold immunochromatographic strip for the rapid detection of Tomato zonate spot tospovirus |
| title_short | Monoclonal antibody-based colloid gold immunochromatographic strip for the rapid detection of Tomato zonate spot tospovirus |
| title_sort | monoclonal antibody based colloid gold immunochromatographic strip for the rapid detection of tomato zonate spot tospovirus |
| topic | Tomato zonate spot virus (TZSV) Monoclonal antibodies Colloid gold immunochromatographic (GICA) strip Rapid detection |
| url | http://link.springer.com/article/10.1186/s12985-018-0919-5 |
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