Development of a sandwich ELISA for the detection of Chinese sacbrood virus infection
Chinese sacbrood disease (CSBD) is a highly pathogenic infectious disease in bees that is caused by Chinese sacbrood virus (CSBV). Although several molecular detection methods have been developed for CSBV, there are no commercially available enzyme-linked immunosorbent assay (ELISA) kits. We therefo...
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Published in | Archives of virology Vol. 165; no. 7; pp. 1551 - 1556 |
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Main Authors | , , , , |
Format | Journal Article |
Language | English |
Published |
Vienna
Springer Vienna
01.07.2020
Springer Nature B.V |
Subjects | |
Online Access | Get full text |
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Summary: | Chinese sacbrood disease (CSBD) is a highly pathogenic infectious disease in bees that is caused by Chinese sacbrood virus (CSBV). Although several molecular detection methods have been developed for CSBV, there are no commercially available enzyme-linked immunosorbent assay (ELISA) kits. We therefore developed a sandwich ELISA to detect CSBV antigens. To this end, monoclonal antibodies were produced using VP2 as an immunogen and subsequently characterized. Hybridomas were screened for the secretion of immunoglobulin G (IgG). Using an unlabeled monoclonal antibody (mAb) for coating and a horseradish peroxidase (HRP)-labeled mAb for detection, a CSBV sandwich ELISA method was established. This method showed specificity for CSBV and did not show cross-reactivity with other bee viruses. The detection limit of the sandwich ELISA was 3.675 × 10
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copies/µL. Sixty bee larvae were tested using our sandwich ELISA method, and the presence of CSBV was verified by reverse transcription polymerase chain reaction (RT-PCR). The total coincidence rate was 90%. Thus, a sandwich ELISA method with high specificity and accuracy and a detection limit of 3.675 × 10
4
copies/µL has been successfully developed and can be used for the clinical detection of CSBV. This method will support rapid diagnosis, real-time monitoring, and early warning of CSBD. |
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Bibliography: | ObjectType-Article-2 SourceType-Scholarly Journals-1 ObjectType-Undefined-1 ObjectType-Feature-3 content type line 23 |
ISSN: | 0304-8608 1432-8798 |
DOI: | 10.1007/s00705-020-04634-2 |