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1.
Chinese Journal of Biotechnology ; (12): 4915-4926, 2023.
Artigo em Chinês | WPRIM | ID: wpr-1008068

RESUMO

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.


Assuntos
Animais , Camundongos , Peste dos Pequenos Ruminantes/prevenção & controle , Anticorpos Monoclonais , Reprodutibilidade dos Testes , Vírus da Peste dos Pequenos Ruminantes , Anticorpos Antivirais , Ensaio de Imunoadsorção Enzimática , Cabras
2.
Chinese Journal of Zoonoses ; (12): 72-80, 2017.
Artigo em Chinês | WPRIM | ID: wpr-511119

RESUMO

In the paper,we introduced the peculiarity of Candida albicans and the disease caused by it,expounded the complexity of the pathogenesis,enumerated the advantages of the RNA-Seq and reviewed its application to study on the pathogenesis of Candida albicans,found out some shortages in previous studies,and anticipated the possible trends of such study in future.In conclusion,some remarkable achievements will bring about by use of improved RNA-Seq for intensive researches on the pathogenesis of Candida albicans.

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