Malachite green(MG), a dye, is an antifungal agent that has been used to treat and prevent fish diseases. It is metabolized into reduced leucomalachite green forms(LMG) that may reside in fish muscles for a long perio...Malachite green(MG), a dye, is an antifungal agent that has been used to treat and prevent fish diseases. It is metabolized into reduced leucomalachite green forms(LMG) that may reside in fish muscles for a long period, thus being harmful to human health. The aim of this study was to develop a competitive and direct enzyme-linked immunosorbent assay(ELISA) to detect MG and LMG specifically. The monoclonal antibody(m Ab) to LMG was generated using a hybridoma technique. The obtained m Ab showed good cross-reactivity(CR) to malachite green(MG), but not to crystal violet(CV) and Brilliant Green(BG). The m Ab was used to develop a fast detecting ELISA of MG and LMG in fish. By introducing the conjugation LMG-HRP, the detection capability was 0.37 ng m L-1 for MG and LMG. The mean recovery from spiked grass carp tissues ranged from 76.2% to 82.9% and the coefficients of variation varied between 1.8% and 7.5%. The stable and efficient monoclonal cell line obtained is a sustainable source of sensitive and specific antibody to MG and LMG.展开更多
AIM:To prospectively assess the sensitivity,specificity and time to positivity of theUltra-rapid urease test (URUT) for Helicobacter pylori(H pylori),and compare the results of one with those of two biopsies. METHODS:...AIM:To prospectively assess the sensitivity,specificity and time to positivity of theUltra-rapid urease test (URUT) for Helicobacter pylori(H pylori),and compare the results of one with those of two biopsies. METHODS:Five antral biopsies were taken in consecutive patients undergoing upper endoscopy:one and two biopsies for URUT,and one each for H pylori culture and histology. URUT was read at 1,5,10,20 and 30 min,1,2,3 and 24 h after biopsy insertion into the reagent.A positive histology and/or culture was used as positive reference“gold standards”. RESULTS:URUT was more sensitive for detecting Hpylori with two biopsies rather than one,at all time points up to 120 min.The sensitivity improved from 3.6% to 82.1% for one biopsy and 10.7% to 85.7% for two biopsies from 1 to 120 min.The sensitivity reached 96.4% at 24 h for both, but the specificity reduced from 100% to 96% and 92% for one and two biopsies,respectively. CONCLUSION:Development of a positive URUT result is hastened by doubling the number of gastric biopsies.We recommend taking two instead of one biopsy to achieve an earlier positive URUT result so that H pylori eradication therapy can be initiated before patient is discharged from the endoscopy suite.展开更多
Viral nervous necrosis （VNN） has been reported to infect larvae and juvenile of humpback grouper, and until now, this virus is one of main problem for humpback grouper. Co-agglutination test proved to be a simple, r...Viral nervous necrosis （VNN） has been reported to infect larvae and juvenile of humpback grouper, and until now, this virus is one of main problem for humpback grouper. Co-agglutination test proved to be a simple, rapid and reliable diagnostic test suitable for use in the field or laboratory without any special apparatus. The study aimed to study application of a co-agglutination test using Staphylococci sensitized with specific antibody for the diagnostic of VNN in grouper. First, brain and eye organ samples from diseased fish are homogenized with phosphate buffer saline （PBS） of pH 7.2. Then, the supernatant is collected after centrifugation at 8,000 rpm for 20 min, and finally one drop of the supematant and one drop of anti VNN antibody sensitized Staphylococci suspension are mixed on a glass slid and observation of the agglutination is performed after 5-10 min. The result shows that co-agglutination technique detects positive VNN in the brain and eye samples. The co-agglutination technique may provide valid result in a very short time as compared with complex method, such as enzyme-linked immunosorbant essay （ELISA） and fluorescient antibody technique （FAT） that require a high cost. Thus, this test can detect VNN faster, simple and more economic.展开更多
Objective：The aim of this study was to prepare monoclonal antibody against P53, a kind of tumor suppressor protein,and use the antibody initial y in clinical immunoassay. Methods：Monoclonal antibody was prepared and...Objective：The aim of this study was to prepare monoclonal antibody against P53, a kind of tumor suppressor protein,and use the antibody initial y in clinical immunoassay. Methods：Monoclonal antibody was prepared and identified via the classic protocol of monoclonal antibody preparation. Identified monoclonal antibodies were purified by af inity chro-matography. Antibody titer was determined by enzyme linked immunosorbent assay （ELISA）. The specific binding activity of antibody was detected by Western blotting and immunohistochemistry. Results：Three strains of monoclonal antibodies named 1P15, 2P37 and 3P40 were obtained and purified by af inity chromatography. The purity of antibodies was higher than 90%. The titers of antibodies were more than 1：6000. Western blot and immunohistochemistry assay showed that the specific antibody can combine with endogenous P53 protein in the tumor celllines and determine the expression of P53 in tumor tis-sue. Conclusion：Three strains of monoclonal antibodies with high af inity to P53 were successful y established, which can be used for detecting the expression of P53 in tumor cells or tissue.展开更多
基金supported by the National High Technology Research and Development Program of China (Granted no. 2011AA10A216)Special Fund for Agroscientific Research in the Public Interest (Granted no. 201203085)
文摘Malachite green(MG), a dye, is an antifungal agent that has been used to treat and prevent fish diseases. It is metabolized into reduced leucomalachite green forms(LMG) that may reside in fish muscles for a long period, thus being harmful to human health. The aim of this study was to develop a competitive and direct enzyme-linked immunosorbent assay(ELISA) to detect MG and LMG specifically. The monoclonal antibody(m Ab) to LMG was generated using a hybridoma technique. The obtained m Ab showed good cross-reactivity(CR) to malachite green(MG), but not to crystal violet(CV) and Brilliant Green(BG). The m Ab was used to develop a fast detecting ELISA of MG and LMG in fish. By introducing the conjugation LMG-HRP, the detection capability was 0.37 ng m L-1 for MG and LMG. The mean recovery from spiked grass carp tissues ranged from 76.2% to 82.9% and the coefficients of variation varied between 1.8% and 7.5%. The stable and efficient monoclonal cell line obtained is a sustainable source of sensitive and specific antibody to MG and LMG.
文摘AIM:To prospectively assess the sensitivity,specificity and time to positivity of theUltra-rapid urease test (URUT) for Helicobacter pylori(H pylori),and compare the results of one with those of two biopsies. METHODS:Five antral biopsies were taken in consecutive patients undergoing upper endoscopy:one and two biopsies for URUT,and one each for H pylori culture and histology. URUT was read at 1,5,10,20 and 30 min,1,2,3 and 24 h after biopsy insertion into the reagent.A positive histology and/or culture was used as positive reference“gold standards”. RESULTS:URUT was more sensitive for detecting Hpylori with two biopsies rather than one,at all time points up to 120 min.The sensitivity improved from 3.6% to 82.1% for one biopsy and 10.7% to 85.7% for two biopsies from 1 to 120 min.The sensitivity reached 96.4% at 24 h for both, but the specificity reduced from 100% to 96% and 92% for one and two biopsies,respectively. CONCLUSION:Development of a positive URUT result is hastened by doubling the number of gastric biopsies.We recommend taking two instead of one biopsy to achieve an earlier positive URUT result so that H pylori eradication therapy can be initiated before patient is discharged from the endoscopy suite.
文摘Viral nervous necrosis （VNN） has been reported to infect larvae and juvenile of humpback grouper, and until now, this virus is one of main problem for humpback grouper. Co-agglutination test proved to be a simple, rapid and reliable diagnostic test suitable for use in the field or laboratory without any special apparatus. The study aimed to study application of a co-agglutination test using Staphylococci sensitized with specific antibody for the diagnostic of VNN in grouper. First, brain and eye organ samples from diseased fish are homogenized with phosphate buffer saline （PBS） of pH 7.2. Then, the supernatant is collected after centrifugation at 8,000 rpm for 20 min, and finally one drop of the supematant and one drop of anti VNN antibody sensitized Staphylococci suspension are mixed on a glass slid and observation of the agglutination is performed after 5-10 min. The result shows that co-agglutination technique detects positive VNN in the brain and eye samples. The co-agglutination technique may provide valid result in a very short time as compared with complex method, such as enzyme-linked immunosorbant essay （ELISA） and fluorescient antibody technique （FAT） that require a high cost. Thus, this test can detect VNN faster, simple and more economic.
基金Supported by grants from the National Natural Science Foundation of China(No.30973562)National Basic Research Program of China(No.2010CB933904)
文摘Objective：The aim of this study was to prepare monoclonal antibody against P53, a kind of tumor suppressor protein,and use the antibody initial y in clinical immunoassay. Methods：Monoclonal antibody was prepared and identified via the classic protocol of monoclonal antibody preparation. Identified monoclonal antibodies were purified by af inity chro-matography. Antibody titer was determined by enzyme linked immunosorbent assay （ELISA）. The specific binding activity of antibody was detected by Western blotting and immunohistochemistry. Results：Three strains of monoclonal antibodies named 1P15, 2P37 and 3P40 were obtained and purified by af inity chromatography. The purity of antibodies was higher than 90%. The titers of antibodies were more than 1：6000. Western blot and immunohistochemistry assay showed that the specific antibody can combine with endogenous P53 protein in the tumor celllines and determine the expression of P53 in tumor tis-sue. Conclusion：Three strains of monoclonal antibodies with high af inity to P53 were successful y established, which can be used for detecting the expression of P53 in tumor cells or tissue.
