Preparation and identification of rabbit anti-marmoset IgG-HRP conjugate antibody / 中国比较医学杂志

Objective To purify marmoset serum IgG, prepare and identify the antiserum and the rabbit anti-marmoset antibody IgG-HRP (horseradish peroxidase). Methods Using SDS-PAGE analysis to identify the serum IgG from HiTrapTM Protein G. The antiserum titer was determined by double immunodiffusion assay. The rabbit anti-marmoset IgG was labeled with HRP by improved sodium periodate method. ELISA and western blotting were used to evaluate the concentration and specificity of rabbit anti-marmoset IgG-HRP. Results The purity of purified marmoset serum IgG determined by SDS-PAGE was higher than 95% , and the anti-serum titer of the anti-marmoset IgG polyclonal antibody was 1∶64. The concentration of rabbit anti-marmoset IgG-HRP identified by direct ELISA was 1∶256 000, and that by western-blotting was 1∶15 000, with a strong specificity. Conclusions The IgG-HRP marker antibody is prepared and the specificity and concentration are identified by ELISA and western blotting. It reserves the resources for the detection system of marmoset pathogens and the molecular immunological testing system.

Detection and analysis of hematological and serum biochemical indices of thirty common marmosets / 中国比较医学杂志

Objective To examine hematological and biochemical parameters in 30 common marmosets,count the mean and standard deviations of each index, and analyze significant differences between female and male groups. Additionally,data from marmosets and macaque breeding were compared. Methods Blood was collected through the posterior limb vein while animals were awake. Hematology and serum biochemical indices were then measured with an automatic blood cell analyzer and blood biochemical analyzer, followed by statistical testing. Results No significant differences were measured in hematological indices between male and female groups. There was a significant difference between the female and male group in serum biochemical indices including high-density lipoprotein-cholesterol(HDL-C) and low-density lipoprotein-cholesterol(LDL-C)(P < 0.05). Compared with the foreign marmoset group, HGB, MCHC,NEUT,ALT, AST, and GLOB were visibly increased in the group of marmosets fed by our institution, but in accordance with the data range in rhesus monkeys. Conclusions Hematological and serum biochemical indices of common marmosets have been detected in this study and compared with related data in macaques and marmosets. Our findings provide basic data not only for pharmacological and toxicological studies,but also diagnosis and treatment of diseases.

Effects of transport and storage conditions on the nucleic acid detection for three kinds of mouse RNA viral samples / 中国比较医学杂志

Objective To analyze the effect of transport and storage conditions on the detection of pathogenic nucleic acid MHV, Reo-3, MNV in laboratory mouse cecal contents samples. Methods MHV, Reo-3 and MNV were mixed with mouse cecal contents and used as reference samples,respectively. They were placed in the lysis buffer of RNA extraction reagent(buffer AVL)or normal saline, and stored at 4℃ and room temperature(22℃-25℃). RNA of these samples was extracted at 1,2,3,7,and 14 days. Then the amount of nucleic acid in samples was detected by real-time fluorescence quantitative PCR. Results A greater decrease of the amount of nucleic acid was observed when the samples were placed in normal saline than that kept in buffer AVL. The amount of nucleic acid in samples stored at 4℃ was found to be higher than that stored at 25℃ room temperature. The amount of nucleic acid in the samples which were kept in buffer AVL at 4℃ for 3 days was higher than 50%,still detectable in the samples kept for 7 days,and undetectable at 14 days. Conclusions Mouse cecal content samples are preferably stored in the lysis buffer of RNA extraction reagent and transported at 4℃ for the detection of MHV, Reo-3, and MNV nucleic acid. It is better to complete the detection test within 3 days.