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Establishment of Eu3+ / Sm3+ dual?label time?resolved fluoroimmunoassay for measurement of anti?bodies to hepatitis C virus and its clinical application / 中华核医学与分子影像杂志
Chinese Journal of Nuclear Medicine and Molecular Imaging ; (6): 677-680, 2018.
Artigo em Chinês | WPRIM | ID: wpr-708936
ABSTRACT
Objective To establish a time-resolved fluoroimmunoassay (TRFIA) system for simul-taneous measurement of immunoglobulin (Ig)M and IgG antibodies to HCV. Methods Recombinant HCV antigen was fixed on microtiter plates to detect serum HCV antibodies. Eu3+-labeled anti-human IgM and Sm3+-labeled anti-human IgG were prepared. HCV-IgM and HCV-IgG TRFIA were established using indirect assay and further optimized and evaluated. The one-sided 95th-percentile was used to calculate the cut-off (CO) values. Results Defining 1 sample/ CO (S/ CO) as measuring unit, the detection limit was 0.06 S/ CO for HCV-IgM and 0.15 S/ CO for HCV-IgG. When diluted a strong-positive specimen from 112.5 to 151200, there was a good liner range within 112.5 to 112800 for HCV-IgM and 125 to 16400 for HCV-IgG. The average intra-assay CV of HCV-IgM and HCV-IgG were 3.37% and 3.66%, respectively, and the aver-age inter-assay CV were 6.52% and 6.75%, respectively. Compared with enzyme-linked immunosorbent as-say (ELISA) kits, the positive conformity rate, the negative conformity rate and total conformity rate were 100.0%(20/ 20), 90.0%(18/ 20), 95.0%(38/ 40) for HCV-IgM TRFIA, and were 100.0% (20/ 20), 95. 0%(19/ 20), 97.5%(39/ 40) for HCV-IgG TRFIA, respectively. Additionally, the established HCV-IgM and HCV-IgG assay kits presented good stability with a decline in the value of fluorescence of 11.1%and 9.5% respectively after being stored at 37 ℃ for 7 d. Conclusions The established HCV-IgM and HCV-IgG TRFIA could simultaneously measure HCV-IgM and HCV-IgG antibodies at one step. The method has wider detectable range and may be a more sensitive, stable, and reliable method for diagnosing HCV in-fection.

Texto completo: DisponíveL Índice: WPRIM (Pacífico Ocidental) Idioma: Chinês Revista: Chinese Journal of Nuclear Medicine and Molecular Imaging Ano de publicação: 2018 Tipo de documento: Artigo

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Texto completo: DisponíveL Índice: WPRIM (Pacífico Ocidental) Idioma: Chinês Revista: Chinese Journal of Nuclear Medicine and Molecular Imaging Ano de publicação: 2018 Tipo de documento: Artigo