Highly sensitive antigen detection procedures for the diagnosis of infectious bovine rhinotracheitis: amplified ELISA and reverse passive haemagglutination |
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| Authors: | S Edwards G C Gitao |
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| Institution: | 1. Key Laboratory of Biorheological Science and Technology, Ministry of Education, College of Bioengineering, Chongqing University, Chongqing 400044, China;2. School of Life Science, Chongqing University, Chongqing 400044, China;3. Laboratory Research Center, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400044, China;1. CAS Key Laboratory of Urban Pollutant Conversion, Institute of Urban Environment, Chinese Academy of Sciences, Xiamen, Fujian 361021, PR China;2. University of Chinese Academy of Sciences, Beijing 100049, PR China;3. College of Resources and Environment, Quanzhou Normal University, Quanzhou, Fujian 362000, PR China;4. College of Chemistry & Chemical Engineering, Northwest Normal University, Lanzhou, Gansu 730070, PR China;5. Tianjin University, Tianjin 300072, PR China;1. Food Science and Technology, Department of Food Science and Technology, Islamic Azad University, Science and Research Branch, Tehran, Iran;2. Department of Food Science and Technology, Science and Research Branch, Islamic Azad University, Tehran, Iran;3. Biotechnology Research Center, Pasteur Institute of Iran, Pasteur sq., Pasteur st., Tehran, Iran |
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| Abstract: | The sensitivity of an enzyme-linked immunosorbent assay (ELISA) for the detection of bovid herpesvirus 1 antigen was increased by up to 50-fold using the biotin-avidin interaction to amplify the reaction, when compared with a simple sandwich ELISA. An alternative immunoassay, reverse passive haemagglutination (RPHA), had a similar sensitivity to the amplified ELISA, and was technically simpler to perform. Both the amplified ELISA and the RPHA could detect viral antigen in the nasal secretions of calves undergoing experimental primary infection with the virus from Day 3 to Day 7 after inoculation. Neither assay was as sensitive as virus isolation in cell culture and they failed to detect antigen in virus-positive samples from the calves from 8 days after inoculation, and from vaccinated calves undergoing challenge infection. |
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