Early diagnosis of diseases is critical in its effective management. Traditional disease detection methods require specialized equipment and trained personnel. With the introduction of rapid diagnostic test kits (RDTs...Early diagnosis of diseases is critical in its effective management. Traditional disease detection methods require specialized equipment and trained personnel. With the introduction of rapid diagnostic test kits (RDTs), disease detection has become easier and faster. However, these RDTs have failed to compete with the specialized laboratory equipment due to their high detection limits and false alarm rates. This paper presents a novel method of using carbon nanofibers (CNFs) grown on glass microballoons (NMBs) to achieve ultra-low detection limits in RDTs. The NMBs have millions of nanosized CNFs grown on each microballoon, with each CNF having a strong bonding affinity for antibodies. The NMBs conjugated with secondary antibodies have therefore a significantly higher probability of capturing minute antigen concentrations in solution. Furthermore, the dark color formation at the capture zone makes visual disease detection possible. Human Immunoglobulin G (IgG) was selected as the model analyte to study the performance of NMBs using a sandwich immunoassay protocol. Ultra-low electrical detection limit of (4 pg/ml) and rapid re- sponse (~1 minute) was achieved using this method.展开更多
Mixed anhydride(MA)was used to conjugate ractopamine(RAC)to BSA and obtained artificial antigen BSA-RAC identified by UV and SDS-PAGE.Balb/c mice were immunized with BSA-RAC and hybridoma lines that secrete RAC monocl...Mixed anhydride(MA)was used to conjugate ractopamine(RAC)to BSA and obtained artificial antigen BSA-RAC identified by UV and SDS-PAGE.Balb/c mice were immunized with BSA-RAC and hybridoma lines that secrete RAC monoclonal antibody(mAb)were generated with cell fusion.A ciELISA kit for detection of RAC(RAC-Kit)was developed with RAC mAb and its performance were tested.The results indicated that BSA-RAC was successfully synthesized and its conjugation ratio of RAC to BSA was about 24.5∶1.Three hybridoma lines were filtered and the best one was 4D8-3E11,its affinity constant(Ka)was 1.65×1010 L/mol.The limit of detection of RAC-Kit was 0.5 ng/ml and its detection range was 0.5-184 ng/ml.The mean recoveries of RAC spiked in feed were 85.6% and in swine urine were 88.6%.The precision and accuracy of the assay as determined by inter-assay and intra-assay coefficient variation were below 15%.It had 9.4% cross-reactivity(CR%)to dobutamine and little or no CR to other compounds.The validity of RAC-Kit in 4 ℃ was in 180 d.展开更多
文摘Early diagnosis of diseases is critical in its effective management. Traditional disease detection methods require specialized equipment and trained personnel. With the introduction of rapid diagnostic test kits (RDTs), disease detection has become easier and faster. However, these RDTs have failed to compete with the specialized laboratory equipment due to their high detection limits and false alarm rates. This paper presents a novel method of using carbon nanofibers (CNFs) grown on glass microballoons (NMBs) to achieve ultra-low detection limits in RDTs. The NMBs have millions of nanosized CNFs grown on each microballoon, with each CNF having a strong bonding affinity for antibodies. The NMBs conjugated with secondary antibodies have therefore a significantly higher probability of capturing minute antigen concentrations in solution. Furthermore, the dark color formation at the capture zone makes visual disease detection possible. Human Immunoglobulin G (IgG) was selected as the model analyte to study the performance of NMBs using a sandwich immunoassay protocol. Ultra-low electrical detection limit of (4 pg/ml) and rapid re- sponse (~1 minute) was achieved using this method.
基金Supported by the Key Project of National Science and Technology Surporting Plan during 11th-Five-Year of China(2006BAK02A21/1)~~
文摘Mixed anhydride(MA)was used to conjugate ractopamine(RAC)to BSA and obtained artificial antigen BSA-RAC identified by UV and SDS-PAGE.Balb/c mice were immunized with BSA-RAC and hybridoma lines that secrete RAC monoclonal antibody(mAb)were generated with cell fusion.A ciELISA kit for detection of RAC(RAC-Kit)was developed with RAC mAb and its performance were tested.The results indicated that BSA-RAC was successfully synthesized and its conjugation ratio of RAC to BSA was about 24.5∶1.Three hybridoma lines were filtered and the best one was 4D8-3E11,its affinity constant(Ka)was 1.65×1010 L/mol.The limit of detection of RAC-Kit was 0.5 ng/ml and its detection range was 0.5-184 ng/ml.The mean recoveries of RAC spiked in feed were 85.6% and in swine urine were 88.6%.The precision and accuracy of the assay as determined by inter-assay and intra-assay coefficient variation were below 15%.It had 9.4% cross-reactivity(CR%)to dobutamine and little or no CR to other compounds.The validity of RAC-Kit in 4 ℃ was in 180 d.
