ELISA for detection of variant rabbit haemorrhagic disease virus RHDV2 antigen in liver extracts
| Autor: | Ana Podadera, Raciel Alarcón González, D. del Llano, M. García Ocaña, Kevin P. Dalton, Francisco Parra, J R de los Toyos, Víctor Granda, Inés Nicieza, J.M. Martín Alonso, Rosa Casais, Fernando Vázquez, Jesús Prieto |
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| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Veterinary Medicine 040301 veterinary sciences medicine.drug_class Hemorrhagic Disease Virus Rabbit Enzyme-Linked Immunosorbent Assay Biology Cross Reactions Monoclonal antibody Diagnostic tools Sensitivity and Specificity Virus law.invention 0403 veterinary science Rabbit haemorrhagic disease 03 medical and health sciences Antigen law Virology medicine Animals Antigens Viral Caliciviridae Infections Diagnostic Tests Routine Reproducibility of Results 04 agricultural and veterinary sciences biology.organism_classification Liver Extracts 030104 developmental biology Liver Polyclonal antibodies biology.protein Recombinant DNA |
| Zdroj: | Journal of virological methods. 251 |
| ISSN: | 1879-0984 |
| Popis: | The emergence and rapid spread of variant of the rabbit hemorrhagic disease virus (RHDV2) require new diagnostic tools to ensure that efficient control measures are adopted. In the present study, a specific sandwich enzyme-linked immunosorbent assay (ELISA) for detection of RHDV2 antigens in rabbit liver homogenates, based on the use of an RHDV2-specific monoclonal antibody (Mab) 2D9 for antigen capture and an anti-RHDV2 goat polyclonal antibody (Pab), was developed. This ELISA was able to successfully detect RHDV2 and RHDV2 recombinant virions with high sensitivity (100%) and specificity (97.22%). No cross-reactions were detected with RHDV G1 viruses while low cross-reactivity was detected with one of the RHDVa samples analyzed. The ELISA afforded good repeatability and had high analytical sensitivity as it was able to detect a dilution 1:163,640 (6.10ng/mL) of purified RHDV-N11 VLPs, which contained approximately 3.4×108molecules/mL particles. The reliable discrimination between closely related viruses is crucial to understand the epidemiology and the interaction of co-existing pathogens. In the work described here we design and validate an ELISA for laboratory based, specific, sensitive and reliable detection of RHDVb/RHDV2. This ELISA is a valuable, specific virological tool for monitoring virus circulation, which will permit a better control of this disease. |
| Databáze: | OpenAIRE |
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