Detection of the circulating antigen 14-3-3 protein of Schistosoma japonicum by time-resolved fluoroimmunoassay in rabbits

Chun-Yan Qian,Xu-Ren Yin,Wei Zhang,Jue Zhang,Chuan-Xin Yu,Li-Jun Song,Biao Huang,Jie Wang,Xue-Dan Ke

doi:10.1186/1756-3305-4-95

Chun-Yan Qian, Xu-Ren Yin + Show 7 more

Open Access

https://doi.org/10.1186/1756-3305-4-95

Copy DOI

Journal: Parasites & Vectors	Publication Date: May 28, 2011
Citations: 22	License type: cc-by

Affiliation: Jiangsu Institute of Parasitic Diseases

Abstract

BackgroundSchistosomiasis remains a major public health concern that afflicts millions of people worldwide. Low levels of Schistosoma infection require more sensitive diagnostic methods. In this study, a time-resolved fluoroimmunoassay (TRFIA) was developed for detecting the signal transduction protein 14-3-3, a circulating antigen of Schistosoma japonicum.ResultsThe detection limit of 14-3-3-TRFIA was 0.78 ng/ml, with a linear measurement range from 0.78 to 800 ng/ml. The average intra-assay and inter-assay variability of this TRFIA was 8.9% and 12.2% respectively, and the mean recovery rate ranged from 92.1% to 115.5%. Within the first 21 days post-infection in rabbits, the positive rates of the 14-3-3-TRFIA were distinctly higher compared to ELISA. All these findings illustrate that 14-3-3-TRFIA has a higher detection efficacy and is a good early diagnostic method for active Schistosoma infection.ConclusionsA sandwich TRFIA for detecting the circulating antigen 14-3-3 of S. japonicum has been developed, and has demonstrated to be a good potential diagnostic method for schistosomiasis.

Highlights

Schistosomiasis remains a major public health concern that afflicts millions of people worldwide
For 14-3-3-enzyme-linked immunosorbent assay (ELISA), the calibration curve was linear from 6.25-200 ng/ml before plateauing. All these data indicated that time-resolved fluoroimmunoassay (TRFIA) has a better sensitivity and measurement range than ELISA
The recovery rates of 14-3-3-TRFIA and 14-3-3-ELISA were analyzed by adding recombinant 14-3-3 proteins at two different concentrations (20 and 100 ng/ml) to healthy rabbit serum from Group B

Summary

Introduction

Schistosomiasis remains a major public health concern that afflicts millions of people worldwide. Serum samples were collected at 0, 7, 14, 21, 28 days post-infection from all rabbits and stored at -80°C for subsequent experiments. The detection limit of TRFIA was defined by the fluorescence of the zero calibrator plus two SD, which was repeated 10 times in one experiment.

Results

Conclusion