ABSTRACT
Objective To express EV71 VP1 in prokaryotic expression system,initially evaluate the ability of blocking EV71 infection and the neutralizing activity of its polyclonal antibody.Methods Construct the prokaryotic expression plasmid pET30a (+)-VP1.Induced expression in Transetta (DE3) with IPTG and identified by Western blot.After purified with HisBind Protein Purification Kit,its ability of blocking EV71 infection and the neutralizing activity of its polyclonal antibody were analyzed.Results Plasmid PET-30a(+)-VPI was constructed successfully and the objective protein was expressed effectively.The ELISA titer of the polyclonal antibody was 1:3200 while neutralizing titer was 1:16 and the recombination protein lost the ability of blocking EV71 infection.Conclusion The recombination protein can stimulate mice to produce antibodies and the polyclonal antibody shew neutralizing activity but the recombination protein lost the binding activity to receptors probably due to the wrong advanced structure.
ABSTRACT
Objective To identify the pathogenic microorganism by MALDI-TOF MS.Methods A total of 560 strains were resuscitated,which included 260 gram-positive bacteria strains,180 gram-negative bacteria strains,60 yeast-like-fungi strains and 60 enteropathogenic bacteria strains.Comparing MALDI-TOF MS with Vitek2 Compact,the discordant results were validated by 16S rDNA sequencing.Results Comparing MALDI-TOF MS with Vitek2 Compact,the coincidence rate was 94.6％ (246/260) for gram-positive bacteria,96.7％ (174/180) for gram-negative bacteria,95％ (57/60) for yeast-like-fungi,and 93.3％(56/60) for enteropathogenic bacteria Fifteen strains were validated by 16S rDNA gene sequencing.Comparing with sequencing,the coincidence rate of two methods was 66.7％ ( 10/15 ) for MALDI-TOF MS and 26.7％(4/11 ) for Vitek2 Compact,respectively.Conclusion MALDI-TOF MS shows rapid turnaround time and modest reagent costs,and it will be another effective tool for microorganism diagnosis.