Introduces a new monitoring method in FMS explicated in some detail by means of the MSF(Monitoring System of FMS)under development by the au- thors.In order to push FMS technology forword,enhance machining flexibility...Introduces a new monitoring method in FMS explicated in some detail by means of the MSF(Monitoring System of FMS)under development by the au- thors.In order to push FMS technology forword,enhance machining flexibility and the flexibility of human operaters and equipment in a FMS,the authors have made some breakthroughs in traditional ways of single item,unit monitoring and self-han- dling,and suggested the idea of integrated inspection and put the MSF into more practicability.The working status of FMS can be monitored on the CRT of a micro- computer of the MSF,system troubles will be shown with icons,by the flash of the system characteristic symbol or by alarming,and so on.This explores a new way for FMS inspection in a wholly integrated manner.展开更多
Objective： To optimize the ELISA for the determination of tetrodotoxin. Methods： A competitive enzyme-linked immunosorbent assay （ELISA） was used. In the ELISA, 100 μl antigen （1. 0 μg/ml） was coated on the mi...Objective： To optimize the ELISA for the determination of tetrodotoxin. Methods： A competitive enzyme-linked immunosorbent assay （ELISA） was used. In the ELISA, 100 μl antigen （1. 0 μg/ml） was coated on the microtiter plate for 60 min at 37 C or over night at 4 C. The plate was then washed 3 times with PBS-T for 3-5 s each time. The optimal incubation time for monoclonal antibody （mAb）, goat anti-mice IgG peroxidase conjugate and OPD were 30 min, 20 min and 10 min at 37 C, re- spectively. Results.. The detection limit is 0. 05 ng in each well. The curve was linear for TTX doses be- tween 5-5 000 ng/ml （0. 25-250 ng for every assay）. The linear regress equation was Y = 0. 30 88X-0.17 41 （R=0.99 01）. The average callback for TTX of muscles and gonads were 99.74% and 100.30%, respectively. The sensitivity of optimization ELISA was 5 times than traditional method and the time of 1.8 h were saved. Conclusion： The optimized ELISA is an ideal method for the determination of tetrodotoxin.展开更多
Detecting white spot syndrome virus （WSSV） in shrimp in high efficiency and veracity is important for disease prevention in aquaculture. Antibody-based mieroarray is a novel proteomic technology that can meet the re...Detecting white spot syndrome virus （WSSV） in shrimp in high efficiency and veracity is important for disease prevention in aquaculture. Antibody-based mieroarray is a novel proteomic technology that can meet the requirements. In this study, we developed an antibody microarray for WSSV-detection in a specific and parallel way at multiple samples. First, seven slides each with different modifications were characterized by atomic force microscope, and were compared in the efficiency of immobilizing proteins. Of the seven, 3-dimensional structured agarose gel-modified slides were chosen appropriate for the microarray for having higher signal value and superior spot size. A purified rabbit anti-WSSV antibody was arrayed as the capture antibody of the microarray on the agarose gel-modified slides, and then the mieroarray slides were incubated in the tissue homogenate of sampled shrimp and the antibody-antigen complex was detected by Cy3-conjugated anti-WSSV monoclonal antibody. The results were measured by a laser chipscanner and analyzed with software. To obtain satisfied fluorescence signal intensity, optimal conditions were searched. The detection limit of the antibody microarray for WSSV is 0.62μg/mL, with a woven long shelf life for 6 months at 4℃ or 8 months at -20℃. Furthermore, concordance between antibody microarray and traditional indirect ELISA reached 100% for WSSV detection. These results suggest that the antibody microarray could be served as an effective tool for diagnostic and epidemiological studies of WSSV.展开更多
Objective:The aim of this study was to explore the relationship between follicular carcinoma of thyroid in different risk groups and lymph node micrometastases.Methods:The keratin-19 of negative neck lymph nodes in 83...Objective:The aim of this study was to explore the relationship between follicular carcinoma of thyroid in different risk groups and lymph node micrometastases.Methods:The keratin-19 of negative neck lymph nodes in 83 cases in routine pathological examination,was detected by SP immunohistochemistry using keratin-19 monoclonal antibody to confirm lymph node micrometastases.All of cases are divided into high risk group,middle risk group and low risk group according to factors related prognosis,the relationship between lymph node micrometastases and different risk groups and follow-up visit documents were analyzed.Results:Fifty-eight neck lymph nodes in 16 cases of 83 cases(19.3%) showed positive lymph node micrometastases,and incidence of lymph node micrometastases was 4/39 in low risk group,5/32 in middle risk group and 7/12 in high risk group,respectively.it showed remarkable difference during 3 groups(P < 0.001).Nine patients in 16 cases with positive lymph node micrometastases showed local recurrence and distant metastases,6 patients in 67 cases with negative lymph node micrometastases showed same result(P < 0.001).Conclusion:Lymph node micrometastases in follicular thyroid carcinoma closely correlated to factors related to prognosis.The detection of lymph node micrometastases can directly assistant postoperative treatment and prognosis evaluation to some extent for follicular thyroid carcinoma.展开更多
A monoclonal antibody (MAb) specific for the bluetongue virus (BTV) group specific antigen (VP7) was characterized for its reactivity with purified virus and recombinant BTV VP7 (rVP7) protein and its suitability for ...A monoclonal antibody (MAb) specific for the bluetongue virus (BTV) group specific antigen (VP7) was characterized for its reactivity with purified virus and recombinant BTV VP7 (rVP7) protein and its suitability for use in the sandwich ELISA.The MAb,designated as 5B5 was specific to VP7 and belongs to IgG2a subclass and was selected for the development of the sELISA in this study.The MAb had a titer of 1:25 with BTV and 1:2 with the rVP7 protein.The sELISA is based on capturing of BTV antigen with VP7 specific MAb followed by detection using BTV polyclonal antiserum raised in rabbits.The assay was evaluated with six cell culture adapted serotypes of BTV that have been isolated from India,1,2,15,17,18 and 23.The assay could detect BTV antigen as early as day 8 in blood.It was also successfully applied for the detection of BTV group specific antigen in clinical samples of blood,washed RBCs,buffy coat and plasma.A total of 102 field samples from animals,suspected of being infected with BTV,were tested and 29.42% were positive.The blood samples were also amplified in cell culture which improved the sensitivity of the assay.Results confirmed that the sELISA is rapid and specific.展开更多
Pepino mosaic virus （PepMV）, monopartite RNA virus, 6,500 pb, belonging to Flexiviridae and Potexvirus group, is highly infectious and easily transmissible. Its economic impact is major for the tomato producer＇s co...Pepino mosaic virus （PepMV）, monopartite RNA virus, 6,500 pb, belonging to Flexiviridae and Potexvirus group, is highly infectious and easily transmissible. Its economic impact is major for the tomato producer＇s countries. Prevention, based on early virus detection is the only effective control measure. Monoclonal antibodies appeared to be very useful tool. The authors used for the production of monoclonal antibodies hybridomas technique, by fusing spleen cells of immunized BALB/c mice to PepMV and SP2/O cancerous cells. The aim of this work is to produce hybridomas producers of Mab that could be used for ELISA in Morocco. At the same time, these efforts will serve to decrease expenses of producers concerning phytosanitory control. We obtained 16 hybridomas lines producers of Mab specific for PepMV. They were tested for efficiencies in ELISA and two lines were retained for production of Mab on large scale （1B 11-G 10 and 5A l-G5）. Isotyping of these two lines showed that they are belonging to IgG 1 class and easily purified by affinity chromatography in agarose column by protein A. The conjugation of these two antibodies to alkaline phosphatase has been verified by DAS-ELISA. These antibodies will enable to diagnose the disease from infected tomato plants, integrating several serological tests to control it and target the actions of struggles.展开更多
Organic phototransistors based on high-quality 2,8-dichloro-5,11-dihexyl-indolo[3,2-b]carbazo(CHICZ)single crystals show the highest photoresponsivity of 3×10^3 A W^-1, photosensitivity of 2×10^4 and the det...Organic phototransistors based on high-quality 2,8-dichloro-5,11-dihexyl-indolo[3,2-b]carbazo(CHICZ)single crystals show the highest photoresponsivity of 3×10^3 A W^-1, photosensitivity of 2×10^4 and the detectivity can achieve 8.4×10^14 Jones. We also discovered good linear dependence of log(photosensitivity) versus the wavelength when the devices were illuminated with a series of sameintensity but different-wavelength lights. The organic phototransistors based on CHICZ single crystal have potential applications in wavelength-detection.展开更多
Glypican-3(GPC3) is a promising tumor marker for hepatocellular carcinoma(HCC) diagnosis with high sensitivity and specificity.The aim of this study was to establish an immunohistochemical detection method for GPC3 us...Glypican-3(GPC3) is a promising tumor marker for hepatocellular carcinoma(HCC) diagnosis with high sensitivity and specificity.The aim of this study was to establish an immunohistochemical detection method for GPC3 using the 7D11 monoclonal antibody(7D11 mAb) and evaluate its application for HCC diagnosis.The feasibility of the 7D11 mAb was evaluated by immunohistochemistry performed on adjacent normal liver and intrahepatic cholangiocarcinoma(ICC) samples,Furthermore,the serum GPC3 levels were evaluated in 40 HCC patients,7 ICC patients and 50 healthy donors.The results showed that GPC3 was expressed in 85% of HCC tissues(34/40),but was undetectable in ICC tissues and adjacent normal tissues.GPC3 was significantly increased in the serum of HCC patients(17/40,42.5%) but was undetectable in the serum of ICC patients(0/7,0%) and healthy donors(0/50,0%).This prospective study evaluated the clinical usefulness of 7D11 mAb for GPC3 detection in HCC patients.In conclusion,the use of 7D11 mAb might be good for GPC3 large-scale applications for clinical diagnosis of HCC.展开更多
A protein conjugate of streptomycin （streptomycin-bovine serum albumin （BSA） conjugate） was prepared and used as immunogen to produce monoclonal antibodies （MAb）. One hybridoma secreting anti-streptomycin MAb wa...A protein conjugate of streptomycin （streptomycin-bovine serum albumin （BSA） conjugate） was prepared and used as immunogen to produce monoclonal antibodies （MAb）. One hybridoma secreting anti-streptomycin MAb was obtained and then used to produce MAb. The MAb named 13H5 showed the 50% maximal inhibitory concentra- tion （IC50） value of 4.65 ng/ml and the IC20value of 0.21 ng/ml in phosphate buffered saline （PBS）. At optimum con- ditions, an indirect competitive enzyme-linked immunosorbent assay （ELISA） and a colloidal gold-based immuno- chromatographic assay （CGIA） were developed and applied to detect streptomycin residues in milk and swine urine samples. The developed ELISA showed that the minimum detection limit was 2.0 and 1.9 ng/ml for milk and swine urine samples, respectively, without obvious cross-reactivity to other tested antibiotics except dihydrostreptomycin which gave a 118.32% cross reaction value. Milk and swine urine samples spiked with streptomycin at 10, 50, 100 and 200 ng/rnl were analyzed by the established ELISA. The mean recovery of streptomycin was from 81.9% to 105.5% and from 84.3% to 92.2% for milk and swine urine, respectively. The optimized CGIA showed that the minimum de- tection limit was 20.0 ng/ml for milk and swine urine samples. The results of spiked analysis and specific analysis demonstrate that the CGIA could be applicable for screening milk and swine urine samples for the presence of streptomycin residues on-site. The established ELISA and CGIA allow the rapid, low-cost, and sensitive determination of streptomycin residues in food samples.展开更多
文摘Introduces a new monitoring method in FMS explicated in some detail by means of the MSF(Monitoring System of FMS)under development by the au- thors.In order to push FMS technology forword,enhance machining flexibility and the flexibility of human operaters and equipment in a FMS,the authors have made some breakthroughs in traditional ways of single item,unit monitoring and self-han- dling,and suggested the idea of integrated inspection and put the MSF into more practicability.The working status of FMS can be monitored on the CRT of a micro- computer of the MSF,system troubles will be shown with icons,by the flash of the system characteristic symbol or by alarming,and so on.This explores a new way for FMS inspection in a wholly integrated manner.
基金the grants from PhD Priming Foundation of Jilin University(430505010276)
文摘Objective： To optimize the ELISA for the determination of tetrodotoxin. Methods： A competitive enzyme-linked immunosorbent assay （ELISA） was used. In the ELISA, 100 μl antigen （1. 0 μg/ml） was coated on the microtiter plate for 60 min at 37 C or over night at 4 C. The plate was then washed 3 times with PBS-T for 3-5 s each time. The optimal incubation time for monoclonal antibody （mAb）, goat anti-mice IgG peroxidase conjugate and OPD were 30 min, 20 min and 10 min at 37 C, re- spectively. Results.. The detection limit is 0. 05 ng in each well. The curve was linear for TTX doses be- tween 5-5 000 ng/ml （0. 25-250 ng for every assay）. The linear regress equation was Y = 0. 30 88X-0.17 41 （R=0.99 01）. The average callback for TTX of muscles and gonads were 99.74% and 100.30%, respectively. The sensitivity of optimization ELISA was 5 times than traditional method and the time of 1.8 h were saved. Conclusion： The optimized ELISA is an ideal method for the determination of tetrodotoxin.
基金Supported by the National High Technology Research and Development Program of China (863 Program) (No. 2006AA100306)Special Fund for Agro-Scientific Research in the Public Interest (No. 201103034)
文摘Detecting white spot syndrome virus （WSSV） in shrimp in high efficiency and veracity is important for disease prevention in aquaculture. Antibody-based mieroarray is a novel proteomic technology that can meet the requirements. In this study, we developed an antibody microarray for WSSV-detection in a specific and parallel way at multiple samples. First, seven slides each with different modifications were characterized by atomic force microscope, and were compared in the efficiency of immobilizing proteins. Of the seven, 3-dimensional structured agarose gel-modified slides were chosen appropriate for the microarray for having higher signal value and superior spot size. A purified rabbit anti-WSSV antibody was arrayed as the capture antibody of the microarray on the agarose gel-modified slides, and then the mieroarray slides were incubated in the tissue homogenate of sampled shrimp and the antibody-antigen complex was detected by Cy3-conjugated anti-WSSV monoclonal antibody. The results were measured by a laser chipscanner and analyzed with software. To obtain satisfied fluorescence signal intensity, optimal conditions were searched. The detection limit of the antibody microarray for WSSV is 0.62μg/mL, with a woven long shelf life for 6 months at 4℃ or 8 months at -20℃. Furthermore, concordance between antibody microarray and traditional indirect ELISA reached 100% for WSSV detection. These results suggest that the antibody microarray could be served as an effective tool for diagnostic and epidemiological studies of WSSV.
文摘Objective:The aim of this study was to explore the relationship between follicular carcinoma of thyroid in different risk groups and lymph node micrometastases.Methods:The keratin-19 of negative neck lymph nodes in 83 cases in routine pathological examination,was detected by SP immunohistochemistry using keratin-19 monoclonal antibody to confirm lymph node micrometastases.All of cases are divided into high risk group,middle risk group and low risk group according to factors related prognosis,the relationship between lymph node micrometastases and different risk groups and follow-up visit documents were analyzed.Results:Fifty-eight neck lymph nodes in 16 cases of 83 cases(19.3%) showed positive lymph node micrometastases,and incidence of lymph node micrometastases was 4/39 in low risk group,5/32 in middle risk group and 7/12 in high risk group,respectively.it showed remarkable difference during 3 groups(P < 0.001).Nine patients in 16 cases with positive lymph node micrometastases showed local recurrence and distant metastases,6 patients in 67 cases with negative lymph node micrometastases showed same result(P < 0.001).Conclusion:Lymph node micrometastases in follicular thyroid carcinoma closely correlated to factors related to prognosis.The detection of lymph node micrometastases can directly assistant postoperative treatment and prognosis evaluation to some extent for follicular thyroid carcinoma.
文摘A monoclonal antibody (MAb) specific for the bluetongue virus (BTV) group specific antigen (VP7) was characterized for its reactivity with purified virus and recombinant BTV VP7 (rVP7) protein and its suitability for use in the sandwich ELISA.The MAb,designated as 5B5 was specific to VP7 and belongs to IgG2a subclass and was selected for the development of the sELISA in this study.The MAb had a titer of 1:25 with BTV and 1:2 with the rVP7 protein.The sELISA is based on capturing of BTV antigen with VP7 specific MAb followed by detection using BTV polyclonal antiserum raised in rabbits.The assay was evaluated with six cell culture adapted serotypes of BTV that have been isolated from India,1,2,15,17,18 and 23.The assay could detect BTV antigen as early as day 8 in blood.It was also successfully applied for the detection of BTV group specific antigen in clinical samples of blood,washed RBCs,buffy coat and plasma.A total of 102 field samples from animals,suspected of being infected with BTV,were tested and 29.42% were positive.The blood samples were also amplified in cell culture which improved the sensitivity of the assay.Results confirmed that the sELISA is rapid and specific.
文摘Pepino mosaic virus （PepMV）, monopartite RNA virus, 6,500 pb, belonging to Flexiviridae and Potexvirus group, is highly infectious and easily transmissible. Its economic impact is major for the tomato producer＇s countries. Prevention, based on early virus detection is the only effective control measure. Monoclonal antibodies appeared to be very useful tool. The authors used for the production of monoclonal antibodies hybridomas technique, by fusing spleen cells of immunized BALB/c mice to PepMV and SP2/O cancerous cells. The aim of this work is to produce hybridomas producers of Mab that could be used for ELISA in Morocco. At the same time, these efforts will serve to decrease expenses of producers concerning phytosanitory control. We obtained 16 hybridomas lines producers of Mab specific for PepMV. They were tested for efficiencies in ELISA and two lines were retained for production of Mab on large scale （1B 11-G 10 and 5A l-G5）. Isotyping of these two lines showed that they are belonging to IgG 1 class and easily purified by affinity chromatography in agarose column by protein A. The conjugation of these two antibodies to alkaline phosphatase has been verified by DAS-ELISA. These antibodies will enable to diagnose the disease from infected tomato plants, integrating several serological tests to control it and target the actions of struggles.
基金financial support from the Ministry of Science and Technology of China (2017YFA0204503 and 2016YFB0401100)the National Natural Science Foundation of China (51725304, 51633006, 51703159 and 51733004)the Strategic Priority Research Program (XDB12030300) of the Chinese Academy of Sciences
文摘Organic phototransistors based on high-quality 2,8-dichloro-5,11-dihexyl-indolo[3,2-b]carbazo(CHICZ)single crystals show the highest photoresponsivity of 3×10^3 A W^-1, photosensitivity of 2×10^4 and the detectivity can achieve 8.4×10^14 Jones. We also discovered good linear dependence of log(photosensitivity) versus the wavelength when the devices were illuminated with a series of sameintensity but different-wavelength lights. The organic phototransistors based on CHICZ single crystal have potential applications in wavelength-detection.
基金supported by the National High Technology Research and Development Program of China (2012AA020205)the Union Program of the Education Ministry,Guangdong Province (2011A090200028)the Combination Project of the Education Ministry,Guangdong Province (2010B090400422)
文摘Glypican-3(GPC3) is a promising tumor marker for hepatocellular carcinoma(HCC) diagnosis with high sensitivity and specificity.The aim of this study was to establish an immunohistochemical detection method for GPC3 using the 7D11 monoclonal antibody(7D11 mAb) and evaluate its application for HCC diagnosis.The feasibility of the 7D11 mAb was evaluated by immunohistochemistry performed on adjacent normal liver and intrahepatic cholangiocarcinoma(ICC) samples,Furthermore,the serum GPC3 levels were evaluated in 40 HCC patients,7 ICC patients and 50 healthy donors.The results showed that GPC3 was expressed in 85% of HCC tissues(34/40),but was undetectable in ICC tissues and adjacent normal tissues.GPC3 was significantly increased in the serum of HCC patients(17/40,42.5%) but was undetectable in the serum of ICC patients(0/7,0%) and healthy donors(0/50,0%).This prospective study evaluated the clinical usefulness of 7D11 mAb for GPC3 detection in HCC patients.In conclusion,the use of 7D11 mAb might be good for GPC3 large-scale applications for clinical diagnosis of HCC.
基金Project (No.2007C22047) supported by the Program of Science and Technology of Zhejiang Province,China
文摘A protein conjugate of streptomycin （streptomycin-bovine serum albumin （BSA） conjugate） was prepared and used as immunogen to produce monoclonal antibodies （MAb）. One hybridoma secreting anti-streptomycin MAb was obtained and then used to produce MAb. The MAb named 13H5 showed the 50% maximal inhibitory concentra- tion （IC50） value of 4.65 ng/ml and the IC20value of 0.21 ng/ml in phosphate buffered saline （PBS）. At optimum con- ditions, an indirect competitive enzyme-linked immunosorbent assay （ELISA） and a colloidal gold-based immuno- chromatographic assay （CGIA） were developed and applied to detect streptomycin residues in milk and swine urine samples. The developed ELISA showed that the minimum detection limit was 2.0 and 1.9 ng/ml for milk and swine urine samples, respectively, without obvious cross-reactivity to other tested antibiotics except dihydrostreptomycin which gave a 118.32% cross reaction value. Milk and swine urine samples spiked with streptomycin at 10, 50, 100 and 200 ng/rnl were analyzed by the established ELISA. The mean recovery of streptomycin was from 81.9% to 105.5% and from 84.3% to 92.2% for milk and swine urine, respectively. The optimized CGIA showed that the minimum de- tection limit was 20.0 ng/ml for milk and swine urine samples. The results of spiked analysis and specific analysis demonstrate that the CGIA could be applicable for screening milk and swine urine samples for the presence of streptomycin residues on-site. The established ELISA and CGIA allow the rapid, low-cost, and sensitive determination of streptomycin residues in food samples.
