Study On Specific Detection Of Herpes Simplex Virus In Subgingival Plague By Magnetic Upconversion Microspheres

Objectives Porous carboxylated polystyrene microspheres were used to embed magnetic up-conversion microspheres of upconversion nanoparticles(UCNPs)Na YF4:Yb,Er and magnetic nanoparticles(MNPs).The fluorescence characteristics of up-conversion and the enrichment and separation of magnetic particles from microorganisms.To achieve rapid and accurate detection of Herpes simplex virus 1(HSV-1)in subgingival plaque samples of clinical patients.To quickly test periodontal pathogenic viruses on the chair side useing up-conversion nanomaterials.Provides research basis for the simultaneous detection of multiple periodontal pathogenic factors in the future.Methods Dispersion polymerization method was used to synthesize the polystyrene seeds.Prepare the porous carboxylated polystyrene nanoospheres by Seed Polymerization.Then Na YF4:Yb,Er nanoparticle(UCNPs)and magnetic nanoparticle(MNPs)were converted by high temperature swelling method.MNPs were embedded in porous polystyrene microspheres to obtain magnetically upconverted fluorescent-encoded microspheres with the best magnetic response and fluorescence intensity.Under certain inclusion exclusion criteria,sites with a Probing Depth(PD)greater than or equal to 5 mm in periodontitis were selected and sampled on a sterile paper tip.Microspheres and nested PCR were used to detect the same sample.Monoclonal antibody microspheres attached to HSV-1 were reacted with the sample.Viruses in the sample were detected by double-antibody sandwich assay.The binding of the microspheres to the virus was observed under transmission electron microscope(TEM).Through red and green laser excitation fluorescence microscopy observation test results.Using virus DNA extraction kit,extracting DNA,then perform nested PCR(nested polymerase chain reaction)test,to verify the accuracy of specific microsphere detection system.Results The experimentally produced polystyrene microspheres are uniform in size and have a particle size of about 0.5 ?m.The magnetic particles and up-conversion particles were encapsulated in the high-temperature swelling method to obtain magnetic up-conversion nanoparticles,and the nanoparticles did not have polymerization phenomenon.The microspheres have both magnetic and up-conversion phosphor particle characteristics.The patients who met the standards in the Department of Periodontology from September to December 2017 were collected and saved.The magnetically upconverting nanoparticle has specificity after binding to the monoclonal antibody of HSV-1.After the HSV-1 was ligated,it reacted with the rhodamine-labeled antibody and showed orange fluorescence under a fluorescence microscope.Nested PCR results in the detection of a 232 bp specific band.It has been demonstrated that magnetic up-conversion nanospheres can detect HSV-1 from subgingival plaques.Conclusion1.This project uses high temperature swelling method to embed rare earth upconverting nanoparticles and magnetic nanoparticles into porous polystyrene microspheres.The microspheres were characterized by scanning electron microscopy,their size was uniform,the particle size was 0.5 ?m,and they had good up-conversion and magnetic effects.2.Microspheres can be combined with monoclonal antibodies to form specific microspheres after activation.Microspheres can be specifically bound to HSV-1 after reaction with HSV-1 standard solution.Significant fluorescence manifestations can also be seen.The ball can specifically bind to HSV-1.3.Nested PCR was used as a control.It was verified that after simple sample pretreatment,the magnetic upconversion microsphere detection system constructed in this experiment can be used to detect and detect subgingival plaque.Herpes simplex virus type 1 lays the foundation for the establishment of a liquid-phase chip detection platform for high-throughput detection of multiple periodontal pathogenic factors.