The Immunoperoxidase Technique: Localization of Viral Antigens in Cells

Kurstak, Edouard

doi:10.1016/b978-0-12-470205-9.50018-6

Cited by 23 publications

(16 citation statements)

References 35 publications

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“…The less regular detection of immunoperoxidase staining within salivary glands of A. aegypti mosquitoes during the first week of extrinsic incubation corresponds with the infrequent detection of infectivity in salivary glands of this species at that time (8), in contrast to the regular development of high infectivity titers in salivary glands of Yukon mosquitoes after comparable periods of incubation (7,9). However, several modifications of published immunoperoxidase techniques (5) including purification of antiserum before and after conjugation with peroxidase, the use of sodium deoxycholate to eliminate non-specific nuclear staining, and the substitution of orthotolidine for benzidine to avoid the rapid change to brown of the initial blue color of the stained product were required to adapt this procedure successfully to the demonstration of CE antigen in mosquito tissues (Fig. 4).…”

mentioning

confidence: 77%

“…The immunoperoxidase technique (5) has been used successfully to demonstrate the localization of densonucleosis virus in infected cells of Galleria mellonella moths, and more recently to detect measles antigen in infected human cells (3). This reaction depends on the coupling of horseradish peroxidase to antibodycontaining globulin by means of glutaraldehyde (1).…”

Section: Introductionmentioning

confidence: 99%

“…Detection of CE antigen by immunoperoxidase staining, within the cytoplasm of salivary gland acinar cells of three species of Yukon mosquitoes regularly in salivary glands 4 to 7 days after intrathoracic injection with the Marsh Lake 23 strain, after incubation both at 55 and 70°F, extends the utility of this technique to the intracellular localization of arboviruses in mosquito vectors, following its successful application to detection of densonucleosis virus antigen in moth cells (5) and measles antigen in mammalian cells (3). Immunoperoxidase provides a convenient alternative technique to immunofluorescence, since it circumvents the problem of autofluorescence of chitin throughout mosquito tissues.…”

mentioning

confidence: 97%

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California encephalitis virus development in mosquitoes as revealed by transmission studies, immunoperoxidase staining, and electron microscopy

McLean¹,

Gubash²,

Grass³

et al. 1975

Can. J. Microbiol.

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Isolates of the snowshoe hare subtype of California encephalitis (CE) virus from Yukon mosquitoes during 1972 and 1973 were transmitted by bites of Aedes aegypti mosquitoes after 4 to 5 weeks of extrinsic incubation at 55 degrees F after intrathoracic injection, and the 1973 strain was transmitted after mosquitoes were fed virus and held for 3 to 4 weeks at 75 degrees F. Antigen of a 1971 isolate of CE virus (Marsh Lake 23) was detected in salivary glands of infected mosquitoes by the immunoperoxidase technique, using highly purified antiserum before and after conjugation with horseradish peroxidase, plus the use of orthotolidine as a substitute for benzidine. enveloped virions 45 nm in diameter were observed in thin sections of salivary glands of Culiseta inornata mosquitoes 59 days after intrathoracic injection with the 1971 isolate, afterincubation at 55 degrees F.

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mentioning

confidence: 77%

Section: Introductionmentioning

confidence: 99%

mentioning

confidence: 97%

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California encephalitis virus development in mosquitoes as revealed by transmission studies, immunoperoxidase staining, and electron microscopy

McLean¹,

Gubash²,

Grass³

et al. 1975

Can. J. Microbiol.

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“…Recently improved immunoperoxidase techniques are regarded as a powerful aid in the detection of virus antigens and their antibodies and in the study of virus replication, on both optical and electron microscopic levels, in infected cells (KURSTAK, 1971;KURSTAK and KURSTAK, 1974;MAYR et al, 1977;CHASEY, 1980). These techniques are similar in sensitivity to immunofluorescence (IF) tests and they also offer operational advantages (POSPISIL et al, 1975;ROSSITTER, 1981; PAN et al, 1982).…”

Section: Discussionmentioning

confidence: 99%

Qualitative and Quantitative Detection of Nairobi Sheep Disease Virus Antigen by Immunoperoxidase in BHK‐, Sheep Kidney‐and Tick Cell Cultures

Muñz¹,

Reimann²,

Jager³

1984

Zentralblatt für Veterinärmedizin Reihe B

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Summary The advantages and disadvantages of direct and indirect immunoperoxidase techniques in detecting Nairobi sheep disease virus (NSDV)‐antigen in cell cultures are described. Both techniques are shown to be very advantageous in titrating NSDV in BHK‐cells because of the simplicity of the techniques compared to other methods. Immunoperoxidase techniques can replace NSDV titrations in mice. In secondary and tertiary sheep kidney cells the direct IPT shows an infection in the same way as in BHK‐cells, but this cell type is regarded as less suitable for titration purposes. The indirect IPT is suitable for detection of virus infection in tick cell cultures. Positive immunoperoxidase reactions are independent of the appearance of virus‐induced cytopathic effects in tissue cultures. Zusammenfassung Qualitativer und quantitativer Nachweis von Nairobi Sheep Disease‐Virusantigen mit Immunoperoxydase‐Methoden in BHK‐, Schafnieren‐ und Zeckenzellkulturen Die Ausarbeitung einer direkten und indirekten Immunoperoxidase‐Technik, sowie ihre Vor‐ und Nachteile zum Nachweis von Nairobi Sheep Disease‐Virusantigen in Zellkulturen wird beschrieben. Beide Methoden konnten mit Vorteil eingesetzt werden, um auf einfachere und schnellere Weise als mit anderen Techniken NSDV in BHK‐Zellen zu titrieren. Sie können den Mäuseversuch ersetzen. Die direkte IPT zeigt in ähnlicher Weise wie bei BHK‐Zellen eine Infektion in sekundären und tertiären Schafnierenzellen an, doch wird diese Zellart für Titrationen als weniger geeignet gehalten. Die indirekte IPT ist geeignet, eine Virusinfektion in Zeckenzellkulturen nachzuweisen. Der positive Ausfall der Immunoperoxidase‐Reaktion in den genannten Zellarten ist dabei unabhängig vom Auftreten zytopathischer Veränderungen. Résumé Détection qualitative et quantitative de l'antigène viral de Nairobi Sheep Disease par immunoperoxydase dans des cultures cellulaires BHK, de reins de mouton et de cellules de tique On décrit la préparation d'une technique d'immunoperoxydase directe et indirecte, ses avantages et ses défauts pour la mise en évidence de l'antigène viral de Nairobi Sheep Disease en culture cellulaire. Les deux méthodes ont été avantageuses pour une titration plus simple et plus rapide qu'avec d'autres techniques NSDV dans des cellules BHK. Elles peuvent remplacer l'essai sur des souris. La technique d'immunoperoxydase présente une infection identique dans les cellules BHK comme dans les cellules secondaires et tertiaires rénales de mouton qui cependant sont moins appropriées pour des titrations. La technique d'immunoperoxydase indirecte est valable pour mettre en évidence une infection virale dans des cultures de cellules de tique. Le résultat positif de la réaction d'immunoperoxydase dans les espèces de cellules citíes ne dépend pas de l'apparition de lésions cytopathiques. Resumen Detección cuali y cuantitativa de antígeno virósico de la enfermedad ovina de Nairobi mediante inmunoperoxidasa en cultivos BHK, de riñón de oveja y de células de garrapatas Se describe la elaboración de una técn...

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“…Peroxidase conjugates were formed with 12 parts horse-radish peroxidase and 5 parts serum protein (Kurstak, 1971) after which the complex was purified by the Shabo (1972) sedimentation procedure. Iodination of peroxidase-conjugated globulins was accomplished by the chloramine T method (Hunter, 1967;Yeh et al, 1975).…”

Section: Sera and Serum Conjugationmentioning

confidence: 99%

Comparative studies on ebv antigens by immuno‐fluorescence and immunoperoxidase techniques

Stephens

Traul

Gaudreau

et al. 1977

Intl Journal of Cancer

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Three groups of EBV antigens, VCA, MA and EA, were compared by the techniques of electron microscopic immunoperoxidase (IP) and immunofluorescence (IF). P3HR-1 and EBV superinfected Raji cells served as targets for labelled sera from patients with BL, NPC and IM or from healthy donors. 125I peroxidase-labelled antibodies were also prepared to determine, autoradiographically, the penetration of the complex into the cell system, and to monitor the incubation and washing procedures. The development of a gentle sedimentation technique proved critical in handling the fragile target cells. VCA and MA were readily identified and localized by both procedures without significant modification of the basic techniques. Indentification of early antigens by IP required modification of the fixation method to include a brief treatment in acetone. The diffuse early antigen (EAD) was found to be associated with cellular ribosomes.

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The Immunoperoxidase Technique: Localization of Viral Antigens in Cells

Cited by 23 publications

References 35 publications

California encephalitis virus development in mosquitoes as revealed by transmission studies, immunoperoxidase staining, and electron microscopy

California encephalitis virus development in mosquitoes as revealed by transmission studies, immunoperoxidase staining, and electron microscopy

Qualitative and Quantitative Detection of Nairobi Sheep Disease Virus Antigen by Immunoperoxidase in BHK‐, Sheep Kidney‐and Tick Cell Cultures

Comparative studies on ebv antigens by immuno‐fluorescence and immunoperoxidase techniques

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