Classification of adenoviral conjunctivitis based on sequencing and phylogenetic analysis of hexon and fiber / 中华实验眼科杂志

Objective:To classify human adenovirus (HAdV) of adenoviral conjunctivitis by sequencing and phylogenetic analysis of hexon protein and fiber protein.Methods:A total of 256 conjunctival swabs were collected from the inferior conjunctival sac of 256 patients with viral conjunctivitis in Shanghai from January 2015 to August 2017.After DNA extraction, the whole length of hexon and fiber was amplified by PCR to perform gene sequencing and phylogenetic analysis.The study protocol was approved by the Ethics Committee of Shanghai General Hospital (No.2020-202).Written informed consent was obtained from each subject.Results:Among the samples, 89(34.76%) were positive for hexon gene amplification, including 1(1.12%) of HAdV-C1, 7(7.87%) of HAdV-C2, 20(22.47%) of HAdV-B3, 6(6.747%) of HAdV-E4, 23(25.84%) of HAdV-D8, 17(19.10%) of HAdV-D19, and 15(16.85%) of HAdV-D37.In phylogenetic analysis, sequenced hexon gene was clustered with the reference prototype correctly.In fiber phylogenetic analysis, 15 of HAdV-D19 and 1 of HAdV-D37 were cross clustered.Conclusions:The combined sequencing of hexon and fiber can provide abundant and effective biological information for the subtype and pathogenicity analysis of HAdV.

Co-expression, purification and bioassay of three avian viral antigens / 生物工程学报

To achieve uniform soluble expression of multiple proteins in the same Escherichia coli strain, and simplify the process steps of antigen production in genetic engineering subunit multivalent vaccine, we co-expressed three avian virus proteins including the fowl adenovirus serotype 4 (FAdV-4) Fiber-2 protein, infectious bursal disease virus (IBDV) VP2 protein and egg-drop syndrome virus (EDSV) Fiber protein in E. coli BL21(DE3) cells after optimization of gene codon, promoter, and tandem expression order. The purified proteins were analyzed by Western blotting and agar gel precipitation (AGP). The content of the three proteins were well-proportioned after co-expression and the purity of the purified proteins were more than 80%. Western blotting analysis and AGP experiment results show that all the three co-expression proteins had immunoreactivity and antigenicity. It is the first time to achieve the three different avian virus antigens co-expression and co-purification, which simplified the process of antigen production and laid a foundation for the development of genetic engineering subunit multivalent vaccine.