Development and Validation of a Recombinant Envelop Glycoprotein Based Indirect Enzyme-Linked Immunosorbent Assay to Detect the Antibody to REV |
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Authors: | ZHANG Wen-juan PEI Zong-fei SHI Feng NIU Xing NIU Zhong-xiang |
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Affiliation: | [1]College of Veterinary Medicine, Shandong Agricultural University, Tai'an 271018, P.R.China [2]Veterinary Department, Shandong Vocational Animal Science and Veterinary College, Weifang 261000, P.R.China |
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Abstract: | Based on the antigenic analysis of Reticuloendotheliosis virus (REV) envelop glycoprotein (env) protein,an alternative indirect enzyme-linked immunosorbent assay (ELISA) for detection of REV antibody was developed using a truncated env protein of REV produced in Escherichia coli. This assay was validated by comparison with an indirect immunofluorescence assay (IFA) and a REV-based ELISA. The diagnostic sensitivity (DSN),specificity (DSP) and accuracy of the env indirect-ELISA (ienv-ELISA) were 93.7,93.3 and 93.6% compared with IFA on 1380 field serum samples,and 84.8,95.2 and 91.7% compared with the REV-based ELISA on 96 field sera,respectively. Cross-reactivity assay showed that this assay was REV-specific. Repeatability test revealed that the coefficients of variation of positive sera within and between runs were less than 15%. This method is simpler to produce and perform,time-saving and suitable for large scale survey of REV antibody at low cost. |
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Keywords: | REV env protein prokaryotic expression indirect ELISA antibody detection |
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