1.Detection of Treponema pallidum in tissue: a comparative study of the avidin-biotin-peroxidase complex, indirect immunoperoxidase, FTA-ABS complement techniques and the darkfield method.
Won Soo LEE ; Min Geol LEE ; Kee Yang CHUNG ; Jung Bock LEE
Yonsei Medical Journal 1991;32(4):335-341
With 37 formalin-fixed, paraffin embedded specimens from the lesions of 30 patients with primary, secondary or gastric syphilis, we performed avidin-biotin-peroxidase complex (ABC), indirect immunoperoxidase (IIP) and FTA-ABS complement techniques. Darkfield examination was done in 17 skin lesions. The immunoperoxidase technique, especially the ABC technique, revealed higher reactivity than the FTA-ABS complement technique and darkfield examination in detecting Treponema pallidum in tissues. Furthermore, the ABC technique produced less intense nonspecific background staining than the IIP technique. Histologically, most of the treponemes were located in the upper dermis, epidermis and vessel walls in the order named, and rarely in the lower dermis of the syphilitic skin lesions.
Avidin/diagnostic use
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Biotin/diagnostic use
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Comparative Study
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Fluorescent Antibody Technique
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Fluorescent Treponemal Antibody-Absorption Test
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Human
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Immunoenzyme Techniques
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Treponema pallidum/*isolation & purification
2.Optimization of in situ hybridization assay using non-radioactive DNA probes for the detection of canine herpesvirus (CHV) in paraffin-embedded sections.
Journal of Veterinary Science 2004;5(1):71-73
Two non-radioactive probes using digoxigenin or biotin were developed for detecting canine herpesvirus (CHV) and compared for their sensitivities by in situ hybridization (ISH) in formalin fixed, paraffin embedded sections, which has been used routinely in veterinary fields. Sections of the CHV-infected cell preparation were subjected to several different ISH protocols using digoxigenin- or biotin-labeled probe respectively. Results were compared for the hybridization and background signal intensities. The best result was obtained by the optimized ISH protocol using digoxigenin-labeled probe for detection of CHV DNA. The optimized ISH assay, which developed in this study, may be a valid tool for the study of pathogenesis and diagnosis of CHV infection.
Animals
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Biotin/diagnostic use
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Cell Line
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DNA Probes/chemistry/genetics
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DNA, Viral/chemistry/genetics
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Digoxigenin/diagnostic use
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Dog Diseases/diagnosis/*virology
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Dogs
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Herpesviridae Infections/diagnosis/*veterinary/virology
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Herpesvirus 1, Canid/genetics/*isolation&purification
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In Situ Hybridization/methods/*veterinary
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Polymerase Chain Reaction/veterinary
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Sensitivity and Specificity