Résumé
Objective@#To establish a method for detection of human antibodies against monkeypox virus.@*Mothds@#The enzyme linked immunosorbent assay (ELISA) plates were coasted with two monkeypox virus peptides from B21R protein, to establish an indirect ELISA for detecting monkeypox virus IgG antibody. The healthy individuals serum samples, monkeypox virus infected patient serum samples and other virus infected patient sera samples were applied to evaluate specificity of the peptides antigen. The reaction conditions were optimized.@*Results@#Synthesized two peptides from monkeypox virus BR21R protein did not cross react obviously with healthy person serum and other virus infected serum. It was shown that the reaction condition was best with sera dilution at 1∶50 when two combined peptides were coated at 100 ng /well, and second-antibody was diluted at 1∶20 000. At this condition the cut off value of IgG antibody in serum samples for ELISA were A450 reading of 0.393. The detected results of two serum samples collected from the monkeypox patient in Sierra Leone were strongly positive, the titers of IgG antibody in two sera were both 1∶6 400.@*Conclusions@#The indirect ELISA for detection of monkeypox virus infection was established preliminarily which provided useful tools for epidemiological study and diagnosis.
Résumé
Monkeypox is a zoonotic disease caused by monkeypox virus. It is similar to human smallpox, although typically much less serious and limited to human-to-human transmission. Smallpox no longer occurs following its worldwide eradication in 1980, whereas monkeypox still occurs sporadically in parts of Africa. To respond to the outbreak of monkeypox correctly, we need to rely on effective method of virus detection. This review presents the development in monkeypox virus detection method in recent years to provide reference for the situation of potential virus spreading into our country and for preventing and acting correctly against bioterrorism.
Résumé
Objective@#To generate monkeypox virus specific monoclonal antibodies for further establishing monkeypox virus immunofluorescence assay.@*Methods@#Monkeypox virus A29 protein, vaccinia ortholog A27 protein and cowpox ortholog 162 protein were expressed in E. coli BL21 to screen antibodies. Synthetic monkeypox virus A2917 ~ 49 polypeptide was used to immune BALB/c mice. Monkeypox virus monoclonal antibodies were generated through fusion, cloning and screening techniques. Indirect ELISA was performed to test antibodies specificity and subtype.@*Results@#A29, A27 and 162 proteins were highly expressed in E. coli and detected by Western blot. The three his-tagged proteins were purified using His-Bind affinity chromatography column. The purity of the proteins was all more than 90%. And 8 strains monkeypox virus specific monoclonal antibodies were screened by the three purified proteins. Two mAbs of 8 were IgG3 subtype and the rest were IgG1 subtype.@*Conclusions@#Eight strains of monkeypox virus specific monoclonal antibodies were generated, they can be used to further establish monkeypox virus immune immunofluorescence assay.