Zhongguo xue xi chong bing fang zhi za zhi = Chinese journal of schistosomiasis control · 2011
[Application of F-ELISA for immunodiagnosis of schistosomiasis japonica].
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Hua Wan-Quan, Yu Chuan-Xin, Yin Xu-Ren, Qian Chun-Yan
Аннотация
OBJECTIVE: To optimize the experimental conditions of fast enzyme-linked immunosorbent assay (F-ELISA) and evaluate its performance for immunodiagnosis of schistosomiasis japonica.
METHODS: HRP-SPA was used at different concentrations in the assay to determine the best HRP-SPA concentration by comparing the detection accuracy. The working conditions of F-ELISA were optimized by examining the ratio of absorbance values of positive and negative controls with different incubation time of serum samples and HRP-SPA. To evaluate the performance of F-ELISA in diagnosis of schistosomiasis, the detection accuracy of F-ELISA was compared with routine ELISA and dipstick dye immunoassay (DDIA) by testing serum samples from both healthy subjects and those clinically diagnosed with schistosomiasis and clonorchiasis in parallel.
RESULTS: In F-ELISA, the best working concentration of HRP-SPA was 1:2 000 while the optimized incubation condition for serum samples and HRP-SPA was 60 min at 37 degrees C. Using F-ELISA, we identified 93.8% to be schistosomiasis positive among chronic schistosomiasis subjects, 4% within the clonorchiasis group, and a false positive rate of 1.5% in healthy individuals. This diagnostic accuracy for F-ELISA was similar to routine ELISA and DDIA.
CONCLUSIONS: F-ELISA combines the 2-steps of routine ELISA into a single step making the assay process faster and simpler without sacrificing the diagnostic accuracy. With further investigations, F-ELISA as an easy and practical method is promising to be applied for schistosomiasis diagnosis in the field.
METHODS: HRP-SPA was used at different concentrations in the assay to determine the best HRP-SPA concentration by comparing the detection accuracy. The working conditions of F-ELISA were optimized by examining the ratio of absorbance values of positive and negative controls with different incubation time of serum samples and HRP-SPA. To evaluate the performance of F-ELISA in diagnosis of schistosomiasis, the detection accuracy of F-ELISA was compared with routine ELISA and dipstick dye immunoassay (DDIA) by testing serum samples from both healthy subjects and those clinically diagnosed with schistosomiasis and clonorchiasis in parallel.
RESULTS: In F-ELISA, the best working concentration of HRP-SPA was 1:2 000 while the optimized incubation condition for serum samples and HRP-SPA was 60 min at 37 degrees C. Using F-ELISA, we identified 93.8% to be schistosomiasis positive among chronic schistosomiasis subjects, 4% within the clonorchiasis group, and a false positive rate of 1.5% in healthy individuals. This diagnostic accuracy for F-ELISA was similar to routine ELISA and DDIA.
CONCLUSIONS: F-ELISA combines the 2-steps of routine ELISA into a single step making the assay process faster and simpler without sacrificing the diagnostic accuracy. With further investigations, F-ELISA as an easy and practical method is promising to be applied for schistosomiasis diagnosis in the field.