[Objective] This study aimed to increase the sensitivity and specificity of enzyme-linked immunosorbent assay （ELISA） through analyzing the effects of different blocking buffers and sample diluents and their differe...[Objective] This study aimed to increase the sensitivity and specificity of enzyme-linked immunosorbent assay （ELISA） through analyzing the effects of different blocking buffers and sample diluents and their different concentrations on the result of ELISA. [Method] Different types of blocking buffer [casein, gelatin, BSA, goat serum （GS）, horse serum （HS） and rabbit serum （RS）]and sample diluent （PBST, casein, gelatin, BSA, GS, HS and RS） as well as their different concentrations were tested in ELISA to screen the optimal combination of blocking buffer and sample diluent. [Result] The results showed that 2% BSA had better effect on blocking than 1% and 3% BSA, and both 2% and 1% casein had better blocking effect than 3% casein; 8% and 10% RS showed better blocking effects than 6%RS and 7%RS; compared to BSA and casein, RS had the best effect on blocking, and 8% RS performed best as the blocking buffer and sample diluent. [Conclusion] A good combination of blocking buffer and diluent can effectively reduce the non-specific reaction and improve the sensitivity and specificity of ELISA. This study provides an important reference for the development of a perfect ELISA method.展开更多
基金Supported by the Jiangsu Agriculture Sience and Technology Innovation Fund(JASTIF)[CX(13)3076]Jiangsu Provincial Postdoctoral Sustentation Fund(6511201,1202055C)~~
文摘[Objective] This study aimed to increase the sensitivity and specificity of enzyme-linked immunosorbent assay （ELISA） through analyzing the effects of different blocking buffers and sample diluents and their different concentrations on the result of ELISA. [Method] Different types of blocking buffer [casein, gelatin, BSA, goat serum （GS）, horse serum （HS） and rabbit serum （RS）]and sample diluent （PBST, casein, gelatin, BSA, GS, HS and RS） as well as their different concentrations were tested in ELISA to screen the optimal combination of blocking buffer and sample diluent. [Result] The results showed that 2% BSA had better effect on blocking than 1% and 3% BSA, and both 2% and 1% casein had better blocking effect than 3% casein; 8% and 10% RS showed better blocking effects than 6%RS and 7%RS; compared to BSA and casein, RS had the best effect on blocking, and 8% RS performed best as the blocking buffer and sample diluent. [Conclusion] A good combination of blocking buffer and diluent can effectively reduce the non-specific reaction and improve the sensitivity and specificity of ELISA. This study provides an important reference for the development of a perfect ELISA method.
