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MedLine Citation:
PMID:  23537303     Owner:  NLM     Status:  In-Data-Review    
We demonstrate herein a novel time-resolved fluoroimmunoassay (TRFIA) with high sensitivity and wide range for quantitative detection of ANA-Ig(GAM) antibodies using a biotin-avidin amplification system. The immunoassay was conducted by following procedures for a typical sandwich immunoreactions with cell nucleus form Hela and the Eu(3+)-labeled biotin combined with biotinylated mouse anti-human Ig(GAM) served as the solid nuclear antigen for ANA and the tracer, respectively. The sensitivity, specificity, and stability of the kit were evaluated and comparison with the classical enzyme-linked immunosorbent assay (ELISA) kit was also made. The average intra-assay and interassay CVs detected by the established ANA-Ig(GAM) biotin-avidin-TRFIA were 4.21% and 6.34%, respectively. The lower detection limit was 2.24 U/mL, and the mean recovery rate was 100.74%. The good measurable range of the established biotin-avidin-TRFIA was within 1.95-64,000 U/mL, while it was only within 32.5-4000 U/mL using an ELISA kit. The values determined by the biotin-avidin-TRFIA and ELISA correlated well (R2 = 0.989). The positive rate of healthy volunteers and patients with systemic lupus erythematosus (SLE), rheumatoid arthritis (RA), primary biliary cirrhosis (PBC), Sjögren's syndrome (SS), scleroderma, and mixed connective tissue disease (MCTD) was 0, 100%, 18.5%, 100%, 37.9%, 90.9%, and 92%, respectively. We conclude that the biotin-avidin-TRFIA we developed gives promise for greater sensitivity and accurate detection for ANA-Ig(GAM) in diagnosing and monitoring autoimmune disorders.
Jie Liu; Yan Ye; Zhigang Hu; Yaohong Zou; Guoqian Chen; Lei Yu
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Journal of immunoassay & immunochemistry     Volume:  34     ISSN:  1532-4230     ISO Abbreviation:  J Immunoassay Immunochem     Publication Date:  2013 Apr 
Date Detail:
Created Date:  2013-03-29     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  100963688     Medline TA:  J Immunoassay Immunochem     Country:  England    
Other Details:
Languages:  eng     Pagination:  197-207     Citation Subset:  IM    
a Affiliated Wuxi People's Hospital of Nanjing Medical University , Wuxi , China.
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