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 1128 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 .