Abstract
Immunohistochemistry is a technique in which the specific interaction between an immunoglobulin and its homologous antigen is visualized on histological sections by a microscopically detectable label. Generally, the label consists of an enzyme, such as peroxidase, alkaline phosphatase, or glucose oxidase that reacts with an appropriate substrate-chromogen solution to produce a specific color at the site of reaction. Several immunohistochemical techniques have been developed and the most important are schematically represented in Fig. 1. In the direct method, the primary antibody is directly labeled with the enzyme. In the indirect method, an enzyme-labeled secondary antibody is directed against the immunoglobulin type of the animal species in which the primary antibody has been raised. Both methods have a relatively low sensitivity and are therefore not frequently used.
Schematic representation of immunohistochemical techniques. Direct method, indirect method, ([A] left to right) PAP complex procedure, ABC procedure ([B] left to right). A=avidin; B=biotin; C=substrate-chromogen; P=peroxidase.
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References
Sternberger, L. A., Hardy, P. H., Cuculis, J. J., and Meyer, H. G. (1970) The unlabelled antibody-enzyme method of immunohistochemistry. Preparation and properties of soluble antigen-antibody complex (horseradish peroxidase-antihorseradish peroxidase) and its use in identification of spirochetes. J. Histochem. Cytochem. 18, 315–333.
Hsu, S. M., Raine, L., and Fanger, H. (1981) Use of avidin-biotin-peroxidase complex (ABC) in immunoperoxidase techniques: a comparison between ABC and unlabeled antibody (PAP) procedures. J. Histochem. Cytochem. 29, 577–580.
Mukai, K. and Rosai, J. (1980) Applications of immunoperoxidase techniques in surgical pathology, in Progress in Surgical Pathology, vol. 1 (Fenoglio, C. M. and Wolff, M., eds.), Masson, New York, New York, pp. 15–49.
Elias, J. M. (1982) Principles and Techniques in Diagnostic Histopathology. Noyes Publications, Park Ridge, New Jersey.
Lo, S. C., Wear, D. J., Green, S. L., Jones, P. G., and Legier, J. F. (1993) Adult respiratory distress syndrome with or without systemic disease associated with infections due to Mycoplasma fermentans. Clin. Infect. Dis. 17(Suppl. 1), 259–263.
Morita, T., Fukuda, H., Awakura, T., Shimada, A., Umemura, T., Kazama, S., and Yagihashi, T. (1995) Demonstration of Mycoplasma hyorhinis as a possible primary pathogen for porcine otitis media. Vet. Pathol. 32, 107–111.
De las Mulas, J. M., Fernandez, A., Sierra, M. A., Poveda, J. B., Carranza, J., and De las Mulas, M. (1990) Immunohistochemical demonstration of Mycoplasma gallinarum and Mycoplasma gallinaceum in naturally infected hen oviducts. Res. Vet. Sci. 49, 339–345.
Nunoya, T., Yagihashi, T., Tajima, M., and Nagasawa, Y. (1995) Occurrence of keratoconjunctivitis apparently caused by Mycoplasma gallisepticum in layer chickens. Vet. Pathol. 32, 11–18.
Ferronha, M. H., Nunes Petisca, J. L., Sousa Ferreira, H., Machado, M., Regalla, J., and Penha Goncalves, A. (1990) Detection of Mycoplasma mycoides subsp. mycoides immunoreactive sites in pulmonary tissue and sequestra of bovines with contagious pleuropneumonia, in Contagious Bovine Pleuropneumonia (Regalla, J., ed.), Commission of the European Communities, Luxembourg, pp. 17–25.
Scanziani, E., Paltrinieri, S., and Gelmetti, D. (1991) Identificazione immunoistochimica di Mycoplasma mycoides subsp. mycoides. Osservazioni preliminari. Selezione Veterinaria 32, 33–40.
Scanziani, E., Grieco, V., Boldini, M., Giusti, A. M., and Monaci, C. (1994) Use of immunohistochemistry for diagnosis of contagious bovine pleuropneumonia (CBPP). Proceedings of the 10th International Congress of the International Organization for Mycoplasmology, July 19–26, Bordeaux, France, p. 83.
Scanziani, E., Grieco, V., Boldini, M., and Mandelli, G. (1994) Immunohistochemical identification of Mycoplasma mycoides subsp. mycoides in cases of contagious bovine pleuropneumonia (CBPP). Proceedings of the 12th autumn meeting of the European Society of Veterinary Pathology, September 18–22, Mondovì, Italy, p. 130.
Rodriguez, F., Kennedy, S., Bryson, T. D. G., Fernandez, A., and Ball, H. J. (1996) An immunohistochemical method of detecting Mycoplasma species antigens by use of monoclonal antibodies on paraffin sections of pneumonic bovine and caprine lungs. J. Vet. Med. B 43, 429–438
Adegboye, D. S., Rasberry, U., Halbur, P. G., Andrews, J. J., and Rosenbusch, R. F. (1995) Monoclonal antibody-based immunohistochemical technique for the detection of Mycoplasma bovis in formalin-fixed, paraffin-embedded calf lung tissue. J. Vet. Diagn. Invest. 7, 261–265.
Shi, S. R., Gu, J., Kalra, K. L., Chen, T., Cote, R. J., and Taylor, C. R. (1995) Antigen retrieval technique: a novel approach to immunohistochemistry on routinely processed tissue sections. Cell Vision 2, 6–22.
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© 1998 Humana Press Inc., Totowa, NJ
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Scanziani, E. (1998). Immunohistochemical Staining of Fixed Tissues. In: Miles, R., Nicholas, R. (eds) Mycoplasma Protocols. Methods in Molecular Biology™, vol 104. Humana Press. https://doi.org/10.1385/0-89603-525-5:133
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DOI: https://doi.org/10.1385/0-89603-525-5:133
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