[Objective] The research aimed to establish a rapid detection method for Shigella. [Method] Combining immunomagnetic separation technology with ATP bioluminescence technology, a new kind of fast and accurate ATP biolu...[Objective] The research aimed to establish a rapid detection method for Shigella. [Method] Combining immunomagnetic separation technology with ATP bioluminescence technology, a new kind of fast and accurate ATP bioluminescence magnetic enzyme immunoassay technique for Shigella was established. [Result] Using ATP bioluminescence magnetic enzyme immunoassay technique to detect standard solution for Shigella （ATCC 25931 ）, result showed that correlation coefficient between relative light intensity detected by instrument and bacteria concentration detec- ted by culture counting method was 0.981 1. Moreover, relation curve between relative light intensity and Shigella concentration was drawn. [ Conclusion] The method had a high detection speed and accuracy, and could be used for the rapid detection of pathogen in food and environment.展开更多
Enzyme-linked immunosorbent assay(ELISA)provides a convenient way for the detection of viral pathogens.However,conventional ELISA performed on mirowell plates suffers from poor sensitivity,laborious coating and compli...Enzyme-linked immunosorbent assay(ELISA)provides a convenient way for the detection of viral pathogens.However,conventional ELISA performed on mirowell plates suffers from poor sensitivity,laborious coating and complicated blocking procedures.Herein,we designed a sensitive colorimetric immunoassay by taking advantages of the enrichment and isolation ability of magnetic beads(MBs)and the high loading capacity of gold nanoparticles(AuNPs)for detecting respiratory syncytial virus(RSV)as a pathogen model.RSV was selectively captured and preconcentrated from samples with antibodies functionalized MBs,followed by binding with antibodies labeled AuNPs,which carrying a large amount of alkaline phosphatase(ALP)molecules for colorimetric signal amplification by catalyzing the dephosphorylation of non-colored pNPP to generate colored product pNP.After optimizing the experimental conditions based on the principle of low nonspecific signal,low cost,and high sensitivity,the analytical sensitivity of the developed immunoassay can be improved to 0.27 pg/mL,which is over sevenfold higher than that of commercially available RSV ELISA kits(2 pg/mL).In addition,the total assay time was less than 2.5 h without any pretreatment,which is much more rapid than other reported assays.Therefore,the proposed immunoassay holds great promise for the fabrication of rapid,sensitive,and economic method for the viral pathogen detection.展开更多
文摘[Objective] The research aimed to establish a rapid detection method for Shigella. [Method] Combining immunomagnetic separation technology with ATP bioluminescence technology, a new kind of fast and accurate ATP bioluminescence magnetic enzyme immunoassay technique for Shigella was established. [Result] Using ATP bioluminescence magnetic enzyme immunoassay technique to detect standard solution for Shigella （ATCC 25931 ）, result showed that correlation coefficient between relative light intensity detected by instrument and bacteria concentration detec- ted by culture counting method was 0.981 1. Moreover, relation curve between relative light intensity and Shigella concentration was drawn. [ Conclusion] The method had a high detection speed and accuracy, and could be used for the rapid detection of pathogen in food and environment.
基金the National Natural Science Foundation of China(NSFC,No.21535006)the National Basic Research Program of China(973 Program,No.2011CB933600)the Doctoral Scientific Research Foundation(SWU116058).
文摘Enzyme-linked immunosorbent assay(ELISA)provides a convenient way for the detection of viral pathogens.However,conventional ELISA performed on mirowell plates suffers from poor sensitivity,laborious coating and complicated blocking procedures.Herein,we designed a sensitive colorimetric immunoassay by taking advantages of the enrichment and isolation ability of magnetic beads(MBs)and the high loading capacity of gold nanoparticles(AuNPs)for detecting respiratory syncytial virus(RSV)as a pathogen model.RSV was selectively captured and preconcentrated from samples with antibodies functionalized MBs,followed by binding with antibodies labeled AuNPs,which carrying a large amount of alkaline phosphatase(ALP)molecules for colorimetric signal amplification by catalyzing the dephosphorylation of non-colored pNPP to generate colored product pNP.After optimizing the experimental conditions based on the principle of low nonspecific signal,low cost,and high sensitivity,the analytical sensitivity of the developed immunoassay can be improved to 0.27 pg/mL,which is over sevenfold higher than that of commercially available RSV ELISA kits(2 pg/mL).In addition,the total assay time was less than 2.5 h without any pretreatment,which is much more rapid than other reported assays.Therefore,the proposed immunoassay holds great promise for the fabrication of rapid,sensitive,and economic method for the viral pathogen detection.
