AIM: To prepare a kind of magnetic iron-dextran nanopartides that was coated with anti-E.coli O157:H7 IgG, analyze its application conditions, and try to use it to isolate E.coli O157:H7 from foods. METHODS: Magnetic ...AIM: To prepare a kind of magnetic iron-dextran nanopartides that was coated with anti-E.coli O157:H7 IgG, analyze its application conditions, and try to use it to isolate E.coli O157:H7 from foods. METHODS: Magnetic iron-dextran nanopartides were prepared by the reaction of a mixture of ferric and ferrous ions with dextran polymers under alkaline conditions. The particles were coated with antiserum against E.coli O157: H7 by the periodate oxidation-borohydride reduction procedure. The oxidation time, amount of antibody coating the particles, amount of nanoparticles, incubation time and isolation time were varied to determine their effects on recovery of the organisms. Finally, the optimum conditions for isolating E.coli O157:H7 from food samples were established. RESULTS: E.coli O157:H7 can be isolated from samples within 15 min with the sensitivity of 101 CFU/mL or even less. In the presence of 108 CFU/mL of other organisms, the sensitivity is 101-102 CFU/mL. Nonspecific binding of other bacteria to the particles was not observed. Two and a half hours of enrichment is enough for the particles to detect the target from the food samples inoculated with 1 CFU/g. CONCLUSION: Isolation of target bacteria by immuno magnetic nanoparticles is an efficient method with high sensitivity and specificity. The technique is so simple that it can be operated in lab and field even by untrained personnel.展开更多
Cancer metastasis is the leading cause of death in cancer patients worldwide and one of the major challenges in treating cancer.Circulating tumor cells(CTCs)play a pivotal role in cancer metastasis.However,the content...Cancer metastasis is the leading cause of death in cancer patients worldwide and one of the major challenges in treating cancer.Circulating tumor cells(CTCs)play a pivotal role in cancer metastasis.However,the content of CTCs in peripheral blood is minimal,so the detection of CTCs in real samples is extremely challenging.Therefore,efficient enrichment and early detection of CTCs are essential to achieve timely diagnosis of diseases.In this work,we constructed an innovative and sensitive single-nanoparticle collision electrochemistry(SNCE)biosensor for the detection of MCF-7 cells(human breast cancer cells)by immunomagnetic separation technique and liposome signal amplification strategy.Liposomes embedded with platinum nanoparticles(Pt NPs)were used as signal probes,and homemade gold ultramicroelectrodes(Au UME)were used as the working electrodes.The effective collision between Pt NPs and UME would produce distinguishable step-type current.MCF-7 cells were accurately quantified according to the relationship between cell concentration and collision frequency(the number of step-type currents generated per unit time),realizing highly sensitive and specific detection of MCF-7 cells.The SNCE biosensor has a linear range of 10 cells·mL^(-1)to 10^(5) cells·mL^(-1)with a detection limit as low as 5 cells·mL^(-1).In addition,the successful detection of MCF-7 cells in complex samples showed that the SNCE biosensors have great potential for patient sample detection.展开更多
A potential confounding factor in the development and evaluation of biosensors is the diverse nature of the disciplines involved. Biosensor technology involves electrochemistry, microbiology, chemical synthesis, and e...A potential confounding factor in the development and evaluation of biosensors is the diverse nature of the disciplines involved. Biosensor technology involves electrochemistry, microbiology, chemical synthesis, and engineering, among many other disciplines. Biological systems, due to non-homogeneous distribution, are already imprecise compared with other systems, especially food based systems. Inadequate knowledge of the techniques to moderate this leads to ineffective evaluation strategies and potentially halting the pursuit of excellent technology that was merely poorly evaluated. This research was undertaken to evaluate the effect culture age had on the capture efficiency of the electrically active magnetic nanoparticles (EAMNP) using culture as the evaluation tool. The age of culture used for immunomagnetic separation (IMS) over all the experiments was 6 to 18 hours. Ideal culture age range for evaluating biosensors is 4 to 10 hours according to the growth curve for E. coli O157: H7 in trypticase soy broth. This is supported by the statistically significant difference among organisms in groups from 3 to 10 hours old compared with those grouped from 11 to 18 and >19 hours old (α = 0.05, p = 0.001 and p = 0.014 respectively). The two older categories were not different from each other. The capture efficiency in all biosensor analysis will vary less than when culture of only viable cells is the diagnostic tool. This allows a true evaluation of the consistency and accuracy of the method, less hindered by the variation in the ability to culture the organism.展开更多
基金Supported by the National High-technology Research and Development Program of China (863 Program), No. 2003AA302260
文摘AIM: To prepare a kind of magnetic iron-dextran nanopartides that was coated with anti-E.coli O157:H7 IgG, analyze its application conditions, and try to use it to isolate E.coli O157:H7 from foods. METHODS: Magnetic iron-dextran nanopartides were prepared by the reaction of a mixture of ferric and ferrous ions with dextran polymers under alkaline conditions. The particles were coated with antiserum against E.coli O157: H7 by the periodate oxidation-borohydride reduction procedure. The oxidation time, amount of antibody coating the particles, amount of nanoparticles, incubation time and isolation time were varied to determine their effects on recovery of the organisms. Finally, the optimum conditions for isolating E.coli O157:H7 from food samples were established. RESULTS: E.coli O157:H7 can be isolated from samples within 15 min with the sensitivity of 101 CFU/mL or even less. In the presence of 108 CFU/mL of other organisms, the sensitivity is 101-102 CFU/mL. Nonspecific binding of other bacteria to the particles was not observed. Two and a half hours of enrichment is enough for the particles to detect the target from the food samples inoculated with 1 CFU/g. CONCLUSION: Isolation of target bacteria by immuno magnetic nanoparticles is an efficient method with high sensitivity and specificity. The technique is so simple that it can be operated in lab and field even by untrained personnel.
基金supported by the National Natural Science Foundation of China(Nos.22274037,22376055 and 21904032)the Natural Science Foundation of Hubei Province(2022CFB383)。
文摘Cancer metastasis is the leading cause of death in cancer patients worldwide and one of the major challenges in treating cancer.Circulating tumor cells(CTCs)play a pivotal role in cancer metastasis.However,the content of CTCs in peripheral blood is minimal,so the detection of CTCs in real samples is extremely challenging.Therefore,efficient enrichment and early detection of CTCs are essential to achieve timely diagnosis of diseases.In this work,we constructed an innovative and sensitive single-nanoparticle collision electrochemistry(SNCE)biosensor for the detection of MCF-7 cells(human breast cancer cells)by immunomagnetic separation technique and liposome signal amplification strategy.Liposomes embedded with platinum nanoparticles(Pt NPs)were used as signal probes,and homemade gold ultramicroelectrodes(Au UME)were used as the working electrodes.The effective collision between Pt NPs and UME would produce distinguishable step-type current.MCF-7 cells were accurately quantified according to the relationship between cell concentration and collision frequency(the number of step-type currents generated per unit time),realizing highly sensitive and specific detection of MCF-7 cells.The SNCE biosensor has a linear range of 10 cells·mL^(-1)to 10^(5) cells·mL^(-1)with a detection limit as low as 5 cells·mL^(-1).In addition,the successful detection of MCF-7 cells in complex samples showed that the SNCE biosensors have great potential for patient sample detection.
文摘A potential confounding factor in the development and evaluation of biosensors is the diverse nature of the disciplines involved. Biosensor technology involves electrochemistry, microbiology, chemical synthesis, and engineering, among many other disciplines. Biological systems, due to non-homogeneous distribution, are already imprecise compared with other systems, especially food based systems. Inadequate knowledge of the techniques to moderate this leads to ineffective evaluation strategies and potentially halting the pursuit of excellent technology that was merely poorly evaluated. This research was undertaken to evaluate the effect culture age had on the capture efficiency of the electrically active magnetic nanoparticles (EAMNP) using culture as the evaluation tool. The age of culture used for immunomagnetic separation (IMS) over all the experiments was 6 to 18 hours. Ideal culture age range for evaluating biosensors is 4 to 10 hours according to the growth curve for E. coli O157: H7 in trypticase soy broth. This is supported by the statistically significant difference among organisms in groups from 3 to 10 hours old compared with those grouped from 11 to 18 and >19 hours old (α = 0.05, p = 0.001 and p = 0.014 respectively). The two older categories were not different from each other. The capture efficiency in all biosensor analysis will vary less than when culture of only viable cells is the diagnostic tool. This allows a true evaluation of the consistency and accuracy of the method, less hindered by the variation in the ability to culture the organism.
