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 develo...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 1 380 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.展开更多
The envelope protein （Env） of lentiviruses such as HIV, SIV, FIV and EIAV is larger than that of other retroviruses. The Chinese EIAV attenuated vaccine is based on Env and has helped to successfully control this vi...The envelope protein （Env） of lentiviruses such as HIV, SIV, FIV and EIAV is larger than that of other retroviruses. The Chinese EIAV attenuated vaccine is based on Env and has helped to successfully control this virus, demonstrating that envelope is crucial for vaccine. We compared Env variation of the four kinds of lentiviruses. Phylogenetic analysis showed that the evolutionary relationship of Env between HIV and SIV was the closest and they appeared to descend from a common ancestor, and the relationship of HIV and EIAV was the furthest. EIAV had the shortest Env length and the least number of potential N-linked glyeosylation sites （PNGS） as well as glyeosylation density compared to various immunodefieiency viruses. However, HIV had the longest Env length and the most PNGS. Moreover, the alignment of HIV and SIV showed that PNGS were primarily distributed within extraeellular membrane protein gp120 rather than transmembrane gp41. It implies that the size difference among these viruses is associated with a lentivirus specific function and also the diversity of env. There arc low levels of modification of glycosylation sites of Env and selection of optimal protective epitopes might be useful for development of an effective vaccine against HIV/AIDS.展开更多
基金founded by the Key Technologies R&D Program of China during the 11th Five-Year Plan period (2006BAD06A11)
文摘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 1 380 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.
基金Natural Science Foundation of China(30970162)Tianjin Municipal Science and Technology Foundation(08ZCGHHZ01800)
文摘The envelope protein （Env） of lentiviruses such as HIV, SIV, FIV and EIAV is larger than that of other retroviruses. The Chinese EIAV attenuated vaccine is based on Env and has helped to successfully control this virus, demonstrating that envelope is crucial for vaccine. We compared Env variation of the four kinds of lentiviruses. Phylogenetic analysis showed that the evolutionary relationship of Env between HIV and SIV was the closest and they appeared to descend from a common ancestor, and the relationship of HIV and EIAV was the furthest. EIAV had the shortest Env length and the least number of potential N-linked glyeosylation sites （PNGS） as well as glyeosylation density compared to various immunodefieiency viruses. However, HIV had the longest Env length and the most PNGS. Moreover, the alignment of HIV and SIV showed that PNGS were primarily distributed within extraeellular membrane protein gp120 rather than transmembrane gp41. It implies that the size difference among these viruses is associated with a lentivirus specific function and also the diversity of env. There arc low levels of modification of glycosylation sites of Env and selection of optimal protective epitopes might be useful for development of an effective vaccine against HIV/AIDS.
