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Antibody Production for a Rapid Fluorescence Polarization Immunoassay of Estrone
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作者 ZHANG Xuan WANG Qiang +5 位作者 YU Zhong Sergei A. Eremin YU Chun Fai LIU Jin SUN Yuan Ming LEI Hong Tao 《Biomedical and Environmental Sciences》 SCIE CAS CSCD 2014年第1期52-55,共4页
Estrone has been identified as a potential endocrine-disrupting chemical (EDC)[1]. Estrone is usually quantified by gas chromatography-mass spectrometry (GC-MS), GC-MS/MS, high performance liquid chromatography (... Estrone has been identified as a potential endocrine-disrupting chemical (EDC)[1]. Estrone is usually quantified by gas chromatography-mass spectrometry (GC-MS), GC-MS/MS, high performance liquid chromatography (HPLC), HPLC- MS, and HPLC-MS/MS, etc.[2-3]. Meanwhile, several immunoassays based on radioimmunoassay, enzyme linked immunosorbent assay (ELISA) or chemiluminescence immunoassay (CLIA) for determination of estrone in real samples have been developed[2'4]. Although these methods are sensitive, they need multistage separation and are thus time-consuming and laborious. A very promising way for the simplification of immunoassays for routine applications is a shift from heterogeneous methods (with separation) to homogeneous assays (without separation)[5]. Fluorescence polarization immunoassay (FPIA) is one of the homogeneous techniques that meets the requirements of a simple, reliable, fast, and cost-effective analysis[6]. Therefore, the present study is focused on the development of FPIA in order to analyze estrone based on antibody production. 展开更多
关键词 FPIA antibody Production for a Rapid fluorescence Polarization Immunoassay of Estrone
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Cholesterol crystal binding of biliary immunoglobulin A: visualization by fluorescence light microscopy 被引量:6
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作者 Frank Lammert Stefan Südfeld +1 位作者 Norbert Busch Siegfried Matern 《World Journal of Gastroenterology》 SCIE CAS CSCD 2001年第2期198-202,共5页
AIM: To assess potential contributions of biliary IgA for crystal agglomeration into gallstones, we visualized cholesterol crystal binding of biliary IgA. METHODS: Crystal binding biliary proteins were extracted from ... AIM: To assess potential contributions of biliary IgA for crystal agglomeration into gallstones, we visualized cholesterol crystal binding of biliary IgA. METHODS: Crystal binding biliary proteins were extracted from human gallbladder bile using lectin affinity chromatography.Biliary IgA was isolated from the bound protein fraction by immunoaffinity chromatography. Pure cholesterol monohydrate crystals were incubated with biliary IgA and fluoresceine isothiocyanate (FITC)conjugated anti IgA at 37 degree. Samples were examined under polarizing and fluorescence light microscopy with digital image processing. RESULTS: Binding of biliary IgA to cholesterol monohydrate crystals could be visualized with FITC conjugated anti IgA antibodies.Peak fluorescence occurred at crystal edges and dislocations. Controls without biliary IgA or with biliary IgG showed no significant fluorescence. CONCLUSION: Fluorescence light microscopy provided evidence for cholesterol crystal binding of biliary IgA. Cholesterol crystal binding proteins like IgA might be important mediators of crystal agglomeration and growth of cholesterol gallstones by modifying the evolving crystal structures in vivo. 展开更多
关键词 Crystallization Bile CHOLELITHIASIS CHOLESTEROL Chromatography Affinity Fluorescent antibody Technique Humans Image Processing Computer-Assisted Immunoglobulin A Secretory Microscopy fluorescence Research Support Non-U.S. Gov't
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A Fast and Indirect Fluorescent Antibody Assay for the Vibrio in Large Yellow Croaker Pseudosciaena crocea (Richardson) 被引量:2
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作者 王军 苏永全 鄢庆枇 《Chinese Journal of Oceanology and Limnology》 SCIE CAS CSCD 2003年第1期91-96,共6页
A fast and indirect fluorescent antibody assay for the Vibrio alginolyticus and V. parahaemolyticus infecting the large yellow croaker has been developed. The specific antisera for the two strains of vibrio were prepa... A fast and indirect fluorescent antibody assay for the Vibrio alginolyticus and V. parahaemolyticus infecting the large yellow croaker has been developed. The specific antisera for the two strains of vibrio were prepared with New Zealand rabbit and the antiserum and cross-reactive efficacy was tested by coagulation in tube. It showed that the goat anti-rabbit IgG had been labeled by fluorescence isothiocyanate (FITC). The results showed that positive reactions were 100% for the large yellow croaker Pseudosciaena crocea with typical symptom of vibrio infection, while the positive reaction to the pathogen in healthy yellow croakers reached 40%, but seemed negative for aquaculture water. The results demonstrated that this fast and indirect fluorescent antibody assay can be used not only to test the vibrio pathogen in diseased yellow croaker but also in infected animals with no symptom. 展开更多
关键词 Pseudosciaena crocea Vibrio alginolyticus V. Parahaemolyticus indirect fluorescent antibody
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Evaluation of patients with dry eye disease for conjunctival Chlamydia trachomatis and Ureaplasma urealyticum 被引量:6
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作者 Maha Mohssen Abdelfattah Rania Abdelmonem Khattab +1 位作者 Magda H.Mahran Ebrahim S.Elborgy 《International Journal of Ophthalmology(English edition)》 SCIE CAS 2016年第10期1457-1465,共9页
AIM:To determine the possibility of the development of dry eye disease(DED) as a result of persistent infection with Chlamydia trachomatis and Ureaplasma urealyticum in the conjunctiva of patients.METHODS: This st... AIM:To determine the possibility of the development of dry eye disease(DED) as a result of persistent infection with Chlamydia trachomatis and Ureaplasma urealyticum in the conjunctiva of patients.METHODS: This study was conducted on 58 patients of age range 20-50 y,diagnosed with DED confirmed by Schirmer I test and tear breakup time.The non-dry eye control group included 27 subjects of the same age.Ocular specimens were collected as conjunctival scrapings and swabs divided into three groups: the first used for bacterial culture,the second and third taken to detect Chlamydia trachomatis and Ureaplasma urealyticum by direct fluorescent antibody(DFA) assay and polymerase chain reaction(PCR) method. RESULTS: Chlamydia trachomatis was detected in 65.5% and 76% of DED patients by DFA and PCR methods respectively.Ureaplasma urealyticum was found in 44.8% of DED infected patients using the PCR method.Both organisms were identified in only 37.9% of DED patients found to be infected.Control subjects had a 22%detection rate of Chlamydia trachomatis by DFA assay versus a 7% detection rate by PCR; while Ureaplasma urealyticum was detected in 3.7% of the controls by PCR method.The conjunctival culture revealed that gram positive microorganisms represented 75% of isolates with coagulase negative Staphylococci the most common(50%) followed by Staphylococcus aureus(20%),whereas gram negative microorganisms occurred in 25% of cases,isolating Moraxella spp.as the most frequent organism. CONCLUSION: Our results tend to point out that Chlamydia trachomatis and Ureaplasma urealyticum were detected in a moderate percentage of patients with DED,and could be a fair possibility for its development.PCR is more reliable in detecting Chlamydia trachomatis than DFA technique.The presence of isolated conjunctival bacterial microflora can be of some potential value. 展开更多
关键词 dry eye disease CONJUNCTIVA Chlamydia trachomatis Ureaplasma urealyticum direct fluorescent antibody polymerase chain reaction
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Evaluation of a Direct Rapid Immunohistochemical Test (dRIT) for Rapid Diagnosis of Rabies in Animals and Humans 被引量:4
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作者 Shampur Narayan Madhusudana Sundaramurthy Subha +1 位作者 Ullas Thankappan Yajaman Belludi Ashwin 《Virologica Sinica》 SCIE CAS CSCD 2012年第5期299-302,共4页
Presently the gold standard diagnostic technique for rabies is the direct immunofluorescence assay (dFA) which is very expensive and requires a high level of expertise. There is a need for more economical and user f... Presently the gold standard diagnostic technique for rabies is the direct immunofluorescence assay (dFA) which is very expensive and requires a high level of expertise. There is a need for more economical and user friendly tests, particularly for use in developing countries. We have established one such test called the direct rapid immunohistochemical test (dRIT) for diagnosis of rabies using brain tissue. The test is based on capture of rabies nucleoprotein (N) antigen in brain smears using a cocktail of biotinylated monoclonal antibodies specific for the N protein and color development by streptavidin peroxidase-amino ethyl carbazole and counter staining with haematoxollin. The test was done in parallel with standard FAT dFA using 400 brain samples from different animals and humans. The rabies virus N protein appears under fight microscope as reddish brown particles against a light blue background. There was 100 % correlation between the results obtained by the two tests. Also, interpretation of results by dRIT was easier and only required a light microscope. To conclude, this newly developed dRIT technique promises to be a simple, cost effective diagnostic tool for rabies and will have applicability in field conditions prevalent in developing countries. 展开更多
关键词 Rabies Post-mortem diagnosis Fluorescent antibody technique Immunohistochemistry Direct rapid immunohistochemical test(dRIT)
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Preconditioning crush increases the survival rate of motor neurons after spinal root avulsion
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作者 Lin Li Yizhi Zuo Jianwen He 《Neural Regeneration Research》 SCIE CAS CSCD 2014年第5期540-548,共9页
In a previous study, heat shock protein 27 was persistently upregulated in ventral motor neurons following nerve root avulsion or crush. Here, we examined whether the upregulation of heat shock protein 27 would increa... In a previous study, heat shock protein 27 was persistently upregulated in ventral motor neurons following nerve root avulsion or crush. Here, we examined whether the upregulation of heat shock protein 27 would increase the survival rate of motor neurons. Rats were divided into two groups: an avulsion-only group (avtflsion of the L4 lumbar nerve root only) and a crush-avulsion group (the L4 lumbar nerve root was crushed 1 week prior to the avulsion). Immunofluores- cent staining revealed that the survival rate of motor neurons was significantly greater in the crush-avulsion group than in the avulsion-only group, and this difference remained for at least 5 weeks after avulsion. The higher neuronal survival rate may be explained by the upregulation of heat shock protein 27 expression in motor neurons in the crush-avulsion group. Further- more, preconditioning crush greatly attenuated the expression of nitric oxide synthase in the motor neurons. Our findings indicate that the neuroprotective action of preconditioning crush is mediated through the upregulation of heat shock protein 27 expression and the attenuation of neuronal nitric oxide synthase upregulation following avulsion. 展开更多
关键词 nerve regeneration nerve root avulsion spinal nerve root heat shock protein 27 nitric oxide synthase motor neurons fluorescent antibody technique choline acetyltransferase a grant from Education Ministry of Jiangsu Province Excellent Discipline of Jiangsu Province neural regeneration
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Diagnosis,fetal risk and treatment of pemphigoid gestationis in pregnancy:A case report
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作者 Hai-Ning Jiao Ye-Ping Ruan +2 位作者 Yan Liu Meng Pan Hui-Ping Zhong 《World Journal of Clinical Cases》 SCIE 2021年第34期10645-10651,共7页
BACKGROUND Pemphigoid gestationis(PG)is a rare autoimmune blistering disease that usually presents in the second or third trimester,with an incidence of 1 per 50000 pregnancies.PG tends to recur with an earlier onset ... BACKGROUND Pemphigoid gestationis(PG)is a rare autoimmune blistering disease that usually presents in the second or third trimester,with an incidence of 1 per 50000 pregnancies.PG tends to recur with an earlier onset and a more severe course in subsequent pregnancies.Skin biopsy markers can be confirmed by direct immunofluorescence staining.CASE SUMMARY Our patient was diagnosed with PG at 8 mo of gestation with fresh bullous lesion marks on the abdomen and limbs.Termination of the pregnancy was performed by cesarean section at 37+4 wk of gestation.The patient delivered an infant weighing 3620 gm.The infant had urticaria-like and vesicular skin lesions and was diagnosed with PG.The patient was discharged on prednisolone and in a satisfactory condition.The infant was discharged after anti-inflammatory therapy for one week.CONCLUSION PG is a rarely reported disease,and 10%of newborns develop mild clinical symptoms consisting of urticaria-like or vesicular skin lesions.We intend to remind clinicians to consider this condition when a patient presents with such lesions so that treatment can be started early and neonatal morbidity can be taken into account. 展开更多
关键词 Pemphigoid gestationis PREGNANCY NEWBORN PEMPHIGUS Fluorescent antibody techniqu Case report
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Detection of Epstein-Barr virus infection subtype in patients with multiple sclerosis by indirect immunofluorescence assay
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作者 Shan-Chao Zhang Lei Liu +5 位作者 Rui-Jin Wang Hou-Zhen Tuo Yan-Jun Guo Li Yi De-Xin Wang Jia-Wei Wang 《Neuroimmunology and Neuroinflammation》 2014年第1期24-28,共5页
Aim:The aim was to investigate the infectious conditions of Epstein-Barr virus(EBV)in patients with multiple sclerosis(MS).Methods:Cerebrospinal fluid(CSF)of 20 patients with MS and 20 with other neurological diseases... Aim:The aim was to investigate the infectious conditions of Epstein-Barr virus(EBV)in patients with multiple sclerosis(MS).Methods:Cerebrospinal fluid(CSF)of 20 patients with MS and 20 with other neurological diseases(OND)were tested with indirect immunofluorescence for anti-EBV capsid antigen(EBV-CA)immunoglobulin G(IgG),IgG affinity for anti-EBV-CA,anti-EBV-CA immunoglobulin M(IgM),anti-EBV early antigen(EBV-EA)IgG and anti-EBV nuclear antigen(EBNA)IgG.According to the pattern of antibodies in CSF,infection rates of acute,chronic,primary,recurrent,and past infections were analyzed in the two groups of patients.Results:There were no significant differences in anti-EBV-CA,anti-EBC-EA,and anti-EBNA antigen IgG in CSF between MS and OND patients(P>0.05).The positive rate of low affinity for anti-EBV-CA IgG in MS patients was significantly higher than that for OND patients(75%vs.40%,P<0.05).Furthermore,significant differences in the positive rate of anti-EBV-CA IgM were found between MS and OND patients(70%vs.25%,P<0.05).Of the MS patients,75%were in an EBV acute infection state compared with 40%of OND patients(P<0.05).Conclusion:Acute infection of EBV closely correlates with the occurrence of MS. 展开更多
关键词 Epstein-Barr virus fluorescent antibody technique herpes virus 4 multiple sclerosis
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Changes in gap junctional intercellular communication in rabbits lens epithelial cells induced by low power density microwave radiation
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作者 叶娟 姚克 +1 位作者 曾群力 鲁德强 《Chinese Medical Journal》 SCIE CAS CSCD 2002年第12期1873-1876,154,共4页
OBJECTIVE: To demonstrate the changes in gap junctional intercellular communication (GJIC) mediated by low power density microwave radiation in rabbits lens epithelial cells (LECs) and its mechanisms. METHODS: Rabbits... OBJECTIVE: To demonstrate the changes in gap junctional intercellular communication (GJIC) mediated by low power density microwave radiation in rabbits lens epithelial cells (LECs) and its mechanisms. METHODS: Rabbits' eyes were exposed to 5 mW/cm(2) and 10 mW/cm(2) power densities of microwave radiation for 3 hours. The fluorescence-recovery-after-photobleaching (FRAP) method was used to determine the GJIC. The localization and function of connexin 43 in LECs was detected by laser scanning confocal microscopy. RESULTS: The GJIC of rabbits LECs was inhibited by microwave radiation especially in the 10 mW/cm(2) irradiated samples. A decrease in connexin 43-positive staining was seen in 5 mW/cm(2) x 3 h treated LECs. Intracellular space accumulation and cytoplasmic internalization were clearly demonstrated in 10 mW/cm(2) group. CONCLUSIONS: Low power densities microwave radiation (5 mW/cm(2) and 10 mW/cm(2)) induces damage to connexin 43 and inhibits the GJIC of rabbits LECs. These changes result in an osmotic imbalance within the lens and induce early cataract. 5 mW/cm(2) or 10 mW/cm(2) microwave radiation is cataractogenic. 展开更多
关键词 ANIMALS CATARACT Cell Communication Connexin 43 Epithelial Cells Fluorescent antibody Technique Indirect Gap Junctions Lens Crystalline Microwaves RABBITS
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