Preparation of colloidal gold test strips for the detection of antibodies to peste des petits ruminants based on monoclonal antibodies to N protein / 生物工程学报

A simple, fast, and visual method for detecting antibodies against peste des petits ruminants virus (PPRV) using colloidal gold strips was developed. In this study, the pET-32a-N was transformed into Escherichia coli Rosetta (DE3) for expression. Hybridoma cell lines were generated by fusing SP2/0 myeloma cells with splenocytes from immunized mice with the expressed and purified N protein of PPRV. The PPRV N protein was labeled with colloidal gold particles as the gold-labeled antigen. The N protein served as the gold standard antigen and as the test (T) line-coated antigen, while the monoclonal antibody served as the quality control (C) line-coated antibody to assemble the colloidal gold immunochromatographic test strips for detecting antibodies against the N protein of PPRV. Hybridoma cell line designated as 1F1 was able to stably secrete the monoclonal antibody against the N protein of PPRV. The titer of 1F1 monoclonal antibody in ascites was 1:128 000 determined by indirect enzyme-linked immunosorbent assays (ELISA), and the immunoglobulin subtype of the monoclonal antibody was IgG1, with kappa chain. The obtained monoclonal antibody was able to specifically recognize the N protein of PPRV, as shown by Western blotting and indirect immunofluorescent assay (IFA). The developed colloidal gold test strip method was able to detect PPRV antibodies specifically, and there was no difference between different batches of the test strips. Testing of a total of 122 clinical sera showed that the compliance rate of the test strip with ELISA test was 97.6%.The test strip assay developed in this study has good specificity, reproducibility, and sensitivity, and it can be used for the rapid detection of PPRV antibodies.

Characteristics of Proprioception in Patients with Knee Osteoarthritis / 医用生物力学

Objective To explore the proprioception characteristics of knee joints for knee osteoarthritis (KOA) patients before unicondylar knee arthroplasty (UKA) and total knee arthroplasty (TKA). Methods Twenty-nine single-compartment KOA patients were selected. Fifteen patients were treated with UKA (UKA group) and fourteen patients were treated with TKA (TKA group). The test was performed 1 to 3 days before the operation.The keen society scores (KSS) of KOA patients in UKA group and TKA group were compared, and their joint position sense and kinesthesia were compared with control group. Results Significant differences in KSS were found in TKA group and UKA group, and the knee joint position sense and kinesthesia showed no significant differences between TKA group and UKA group.There were no significant differences in the knee joint position sense and kinesthesia between the operated leg and unoperated leg in TKA group and UKA group, and between the left leg and right leg in control group. Compared with control group, there were significant differences between UKA group and TKA group in 60° position sense of the operated leg and unoperated leg.The kinesthesia of the operated leg and unoperated leg in UKA group and TKA group were also significantly different from that in control group. Conclusions Compared with control group, the proprioception of the knee joint in KOA patients was significantly reduced before the surgery, but the characteristics of proprioception in UKA group and TKA group were similar.