Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound   Target Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound

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Query: EC:3.1.3.1 (alkaline phosphatase)
47,916 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

A non-radioactive in situ hybridization (ISH) method was elaborated to detect cytomegalovirus (CMV)-infected cells in tissue specimens processed for diagnostic routine histopathology. A biotinylated CMV-DNA probe was hybridized following a) four different enzymatic predigestions, b) progressively increasing denaturation periods, and then detected by c) streptavidin-biotin, d) a monoclonal antibody against biotin using a three-stage alkaline phosphatase anti-alkaline phosphatase (APAAP)-technique, and e) combining c + d. Autopsy specimens obtained from an infant with acquired CMV-infection, six patients with AIDS, five patients clinically and serologically without CMV-infection, and preoperative needle core biopsies from six renal allografts served as material. ISH was specific and more sensitive when compared to immunohistochemical (IMH) detection of CMV-antigens by a monoclonal antibody. ISH was concluded to be a rapid, practical, and sensitive tool in daily diagnostic histopathology.
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PMID:Detection of cytomegalovirus-infected cells in autopsy material by in situ hybridization. 216 85

In situ hybridisation was performed with a biotinylated DNA probe for herpes simplex virus (HSV) using high temperature denaturation on formalin fixed, paraffin wax sections of lung, brain, ganglion and keratinising and non-keratinising squamous epithelia. Eosinophilic viral nuclear inclusions or characteristically moulded multiple nuclei with altered chromatin, which were present in two cases of HSV encephalitis and one case of viral pneumonitis, all showed complete hybridisation visualised by an alkaline phosphatase/nitroblue tetrazolium detector system. HSV encephalitis and trigeminal ganglionitis, which were confirmed serologically or clinicopathologically but lacked nuclear changes, also gave positive dense nuclear signal in neurons, glias and satellite cells. No staining was present in the ganglion cells in trigeminal zoster, the glia in progressive multifocal leucoencephalopathy, or in a variety of cells in a lung coinfected with cytomegalovirus. In 10 herpetic blisters of squamous epithelia, infected cells hybridised strongly, while morphologically similar herpes zoster lesions remained negative. In neural tissues non-hybridisation staining was most obtrusive in corpora amylacea and seemed to reflect nonspecific probe adherence. In squamous epithelium, major non-hybridisation staining was caused by probe and antibody possibly adhering to intracellular keratin. The HSV probe permits specific detection of virus in the absence of characteristic nuclear changes and allows varicella zoster virus to be differentiated from HSV, provided that the aforementioned problems with non-hybridisation staining are borne in mind.
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PMID:In situ hybridisation in herpetic lesions using a biotinylated DNA probe. 216 33

A dot blot hybridization immunoenzymatic assay for the rapid detection of cytomegalovirus DNA in urine samples was developed by using a digoxigenin-labeled probe which was immunoenzymatically visualized by antidigoxigenin Fab fragments labeled with alkaline phosphatase. A total of 516 urine samples from different groups of subjects were analyzed, and the hybridization assay was able to yield results within 24 h. The results obtained were compared with results for detection of cytomegalovirus antigens in infected cell cultures.
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PMID:Rapid detection of cytomegalovirus DNA in urine samples with a dot blot hybridization immunoenzymatic assay. 217 99

Biliary complications have recently been reported in patients with AIDS. This may take the form of acalculous cholecystitis or more commonly cholangitis, which may or may not be associated with stenosis of the papilla. These conditions must be sought in patients presenting with right hypochondrial pain and elevated alkaline phosphatase. Infection with Cryptosporidium or cytomegalovirus is often associated.
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PMID:[Biliary diseases in acquired immunodeficiency syndrome. Apropos of 2 cases]. 219 80

A 71-yr-old male presented with a 2-month history of fever, malaise, and weight loss. Physical exam revealed chorioretinitis. Laboratory studies were notable for elevated levels of alkaline phosphatase, gamma-glutamyl transpeptidase, aspartate transaminase, and alanine transaminase. Immunoglobulin G antibody to Toxoplasma gondii was positive to a dilution of 1:4096, whereas serologic studies for hepatitis A virus, hepatitis B virus, cytomegalovirus, Epstein-Barr virus, human immunodeficiency virus, Brucella, and Tularemia were negative. A percutaneous biopsy of the liver revealed hepatic granulomas. Culture of the biopsy specimen was negative for growth of mycobacteria or fungi. Spontaneous improvement in clinical and laboratory parameters occurred over a 4-month period.
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PMID:Toxoplasmic chorioretinitis and hepatic granulomas. 222 Jul 41

A method is described for in situ hybridization detection and typing of herpes simplex virus (HSV) using alkaline phosphatase-labeled synthetic oligonucleotide probes in paraffin tissue sections. Sections mounted on slides are prehybridized and denatured before the probe mixture is added. Hybridization proceeds for 1 h at 60 degrees C. Detection of the alkaline phosphatase label is performed using a nitro blue tetrazolium and 5-bromo-4-chloro-3-indolyl phosphate substrate. Specific hybridization with HSV type 1 DNA was found in sections of herpetic esophagitis and encephalitis. There was no discernible background staining. Hybridization with an oligonucleotide probe specific for HSV type 2 was negative. No hybridization occurred to sections of cytomegalovirus- or adenovirus-infected tissue. The development of this technique expands the utility of synthetic oligonucleotide probes to include hybridization reactions in routinely processed and paraffin-embedded tissue. The use of directly labeled oligonucleotide probes for tissue in situ hybridization overcomes problems of probe contamination with vector plasmid DNA, nonspecific avidin binding to tissue, and the danger and inconvenience of working with radioactive materials.
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PMID:Oligonucleotide probe for herpes virus: use in paraffin sections. 223 90

An indirect alkaline phosphatase immunoenzymatic staining method was developed to localise antigens in human embryo fibroblasts that have been induced by cytomegalovirus. The enzyme label was developed with a naphthol salt and fast blue to obtain a bright blue staining of the antigens that could be clearly visualised under an ordinary light microscope. The procedure is rapid, sensitive, and specific and can be used in diagnostic laboratories to detect active infection caused by cytomegalovirus.
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PMID:Alkaline phosphatase immunoenzymatic staining for detection of antigens induced by cytomegalovirus. 241 27

Rapid methods of specific viral diagnosis in formalin fixed, paraffin embedded tissues include identification of viral incusions in routinely stained histologic sections, immunologic staining of viral antigens, and in situ nucleic acid hybridization. To correlate in situ hybridization with immunologic detection methods, sequential two-color staining was used on tissues from 12 patients, each containing characteristic cytomegalovirus (CMV) inclusions, using a biotinylated CMV DNA probe in an avidin-alkaline phosphatase-linked reaction followed by avidin-biotin complex immunoperoxidase staining of CMV antigen. CMV genetic material was seen in all 17 tissues. CMV antigen was detected in 11 of 17 tissues (65%). The DNA hybridization technique provided more intense staining, detected greater numbers of inclusions, and had less background staining than the immunoperoxidase technique. The alkaline phosphatase reaction product was stable through subsequent immunostaining steps, and immunologic reactivity of CMV antigen was not significantly reduced by prior hybridization steps. CMV DNA probe was localized predominantly within cell nuclei, while CMV antigen immunostaining was predominantly cytoplasmic. It was concluded that sequential in situ hybridization and immunocytochemistry can be performed on standard histologic sections. Furthermore, it is likely that the majority of CMV nucleic acid detected by this tissue hybridization technique is unencapsidated, intranuclear viral DNA and not DNA contained within complete CMV nucleocapsids.
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PMID:Cytomegalovirus detection by nonisotopic in situ DNA hybridization and viral antigen immunostaining using a two-color technique. 245 15

Conditions for combination of DNA in situ hybridization, using biotinylated DNA probes, with immunohistochemistry were investigated on cryostat sections, cytological preparations, and paraffin sections. We found that cryostat sections and cytological preparations are suitable for in situ hybridization of target DNA after fixation in acetone, methanol, ethanol, or Carnoy without further proteinase pretreatment. Acetone is also very suitable for immunostaining of cell surface or cytoskeleton antigens. We therefore performed combined immunoenzyme and in situ hybridization staining using this fixative. The best results were obtained when immunoperoxidase staining with diaminobenzidine/H2O2 was followed directly by in situ hybridization. In addition to immunoperoxidase, alkaline phosphatase-antialkaline phosphatase (APAAP) staining with naphthol ASBI phosphate and New Fuchsin as a substrate could be used. In most instances, detection of the biotinylated hybrid with a streptavidin-biotinylated polyalkaline phosphatase method using nitroblue tetrazolium and 5-bromo-4-chloro-3-indolylphosphate as the substrate was preferable. The double stainings were studied on the following test models: (a) frozen tonsil sections: cell surface antigens (pan T) and ribosomal DNA; (b) frozen genital condyloma sections; cytokeratins and human papillomavirus type 6 + 11 (HPV-6/11) DNA; (c) CaSKi cells: cytokeratins and HPV-16 DNA; (d) infected fetal lung fibroblasts: vimentin and cytomegalovirus (CMV) DNA. An adapted procedure was followed on routinely formaldehye-fixed and paraffin-embedded condyloma tissue. Immunoperoxidase staining for papilloma virus capsid antigen could be combined with DNA in situ hybridization with HPV-6/11 DNA. In this model, however, the accessibility of the target DNA had to be improved by enzyme treatment after the immunostaining and before starting the in situ hybridization.
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PMID:Combined immuno- and non-radioactive hybridocytochemistry on cells and tissue sections: influence of fixation, enzyme pre-treatment, and choice of chromogen on detection of antigen and DNA sequences. 246 28

Combined application of a non-radioactive in situ DNA hybridization procedure and the immunoperoxidase technique on one tissue section is described. Of six potential protocols, only one proved to be successful. First, the immunohistochemical procedure including visualization of enzyme activity is performed; the in situ DNA hybridization protocol is then applied. Using this protocol, several antigens, detected with monoclonal antibodies, and target DNAs, detected by using biotinylated human cytomegalovirus or human papilloma virus type 16 DNA probes, could be distinguished by their peroxidase activity (brown precipitate) and alkaline phosphatase activity (purple-blue precipitate) respectively. The method allows immunophenotyping of virus-infected cells as well as simultaneous visualization of two viral parameters. This technique has important implications for research and diagnostic purposes.
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PMID:Simultaneous application of in situ DNA hybridization and immunohistochemistry on one tissue section. 247 18


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