Sources/Clones
Accurate (3F6), Axcel/Accurate (3F6), Biodesign (092, 093), Biogenesis (0G1/1), Biogenex (3F6), Chemicon and Dako (3F6).
Fixation/Preparation
The Dako antibody reacts with an antigenic epitope of humanPneumocystis carinii,which is resistant to fixation in formalin and picric acid, paraffin embedding and extraction with ethanol and xylene. This antibody may also be used to detectP. carinii in smears prepared from bronchoalveolar lavage fluid and sputum samples (Elvin et al, 1988). However, enzymatic digestion of smears (e.g. trypsin) must be performed before staining.
Background
The Dako antibody (IgM, k) reacts with an 82 kD parasite-specific component of humanPneumocystis carinii (Linder et al, 1987). No crossreactivity was found with a number of parasites and fungi (Elvin et al, 1988).
Applications
The explosion in the AIDS epidemic brought about an increased need for specific markers that recognize P. carinii. Newly developed antibodies mark cyst wall and/or trophozoites (Linder and Radio, 1989). While the sensitivity of the immunocytochemical method appears to be greater than the Giemsa stain, it is only slightly better than the GMS stain, warranting the use of immunostaining in sputum, where identification of the pathogen is more difficult than in bronchoalveolar lavage (Linder and Radio, 1989). The other advantage of immunostaining is that only the cyst wall is detected with the silver stain, whilst immunohistochemistry stains both cyst wall and trophozoites (Amin et al, 1992). However, the latter staining pattern may appear amorphous or focally granular, which may be confused with non-specific staining of mucin or intracellular/free particulate material (Amin et al, 1992). The 3F6 monoclonal antibody has been found to be consistently more sensitive at detecting cysts ofPneumocystis in both sputum and bronchoalveolar lavage specimens (Elvin et al, 1988; Wazir et al, 1994b).
Comments
Immunohistochemistry forP. carinii is a useful adjunct to traditional Giemsa and silver stains, particularly in cytopathology laboratories examining a large number of respiratory specimens from HIV-positive patients.
References
•Amin MB, Mezger E, Zarbo RJ 1992. Detection ofPneumocystis carinii.Comparative study of monoclonal antibody and silver staining. American Journal of Clinical Pathology; 98: 13-18.
•Elvin KM, Bjâkman A, Linder E, Heurlin N, Hlorpe A 1988.Pneumocystis cariniipneumonia: detection of parasites in sputum and bronchoalveolar lavage fluid by monoclonal antibodies. British Medical Journal 297: 381-384.
•Linder E, Lundin L, Vorma H 1987. Detection ofPneumocystis cariniiin lung-derived samples using monoclonal antibodies to an 82kDa parasite component. Journal of Immunological Methods 98: 57-62.
•Linder J, Radio SJ 1989. Immunohistochemistry ofPneumocystis carinii.Seminars in Diagnostic Pathology; 6: 238-244. Pneumocystis carinii.Seminars in Diagnostic Pathology; 6: 238-244.
•Wazir JF, Macrorie SG, Coleman DV 1994. Evaluation of the sensitivity, specificity, and predictive value of monoclonal antibody 3F6 for the detection ofPneumocystis cariniipneumonia in bronchoalveolar lavage specimens and induced sputum. Cytopathology; 5: 82-89.
Bibliografia
Manual of diagnostic antibodies for immunohistology / Anthony S.-Y. Leong, Kumarasen Cooper, F. Joel W.-M. Leong.
Accurate (3F6), Axcel/Accurate (3F6), Biodesign (092, 093), Biogenesis (0G1/1), Biogenex (3F6), Chemicon and Dako (3F6).
Fixation/Preparation
The Dako antibody reacts with an antigenic epitope of humanPneumocystis carinii,which is resistant to fixation in formalin and picric acid, paraffin embedding and extraction with ethanol and xylene. This antibody may also be used to detectP. carinii in smears prepared from bronchoalveolar lavage fluid and sputum samples (Elvin et al, 1988). However, enzymatic digestion of smears (e.g. trypsin) must be performed before staining.
Background
The Dako antibody (IgM, k) reacts with an 82 kD parasite-specific component of humanPneumocystis carinii (Linder et al, 1987). No crossreactivity was found with a number of parasites and fungi (Elvin et al, 1988).
Applications
The explosion in the AIDS epidemic brought about an increased need for specific markers that recognize P. carinii. Newly developed antibodies mark cyst wall and/or trophozoites (Linder and Radio, 1989). While the sensitivity of the immunocytochemical method appears to be greater than the Giemsa stain, it is only slightly better than the GMS stain, warranting the use of immunostaining in sputum, where identification of the pathogen is more difficult than in bronchoalveolar lavage (Linder and Radio, 1989). The other advantage of immunostaining is that only the cyst wall is detected with the silver stain, whilst immunohistochemistry stains both cyst wall and trophozoites (Amin et al, 1992). However, the latter staining pattern may appear amorphous or focally granular, which may be confused with non-specific staining of mucin or intracellular/free particulate material (Amin et al, 1992). The 3F6 monoclonal antibody has been found to be consistently more sensitive at detecting cysts ofPneumocystis in both sputum and bronchoalveolar lavage specimens (Elvin et al, 1988; Wazir et al, 1994b).
Comments
Immunohistochemistry forP. carinii is a useful adjunct to traditional Giemsa and silver stains, particularly in cytopathology laboratories examining a large number of respiratory specimens from HIV-positive patients.
References
•Amin MB, Mezger E, Zarbo RJ 1992. Detection ofPneumocystis carinii.Comparative study of monoclonal antibody and silver staining. American Journal of Clinical Pathology; 98: 13-18.
•Elvin KM, Bjâkman A, Linder E, Heurlin N, Hlorpe A 1988.Pneumocystis cariniipneumonia: detection of parasites in sputum and bronchoalveolar lavage fluid by monoclonal antibodies. British Medical Journal 297: 381-384.
•Linder E, Lundin L, Vorma H 1987. Detection ofPneumocystis cariniiin lung-derived samples using monoclonal antibodies to an 82kDa parasite component. Journal of Immunological Methods 98: 57-62.
•Linder J, Radio SJ 1989. Immunohistochemistry ofPneumocystis carinii.Seminars in Diagnostic Pathology; 6: 238-244. Pneumocystis carinii.Seminars in Diagnostic Pathology; 6: 238-244.
•Wazir JF, Macrorie SG, Coleman DV 1994. Evaluation of the sensitivity, specificity, and predictive value of monoclonal antibody 3F6 for the detection ofPneumocystis cariniipneumonia in bronchoalveolar lavage specimens and induced sputum. Cytopathology; 5: 82-89.
Bibliografia
Manual of diagnostic antibodies for immunohistology / Anthony S.-Y. Leong, Kumarasen Cooper, F. Joel W.-M. Leong.
