+ Site Statistics
+ Search Articles
+ Subscribe to Site Feeds
EurekaMag Most Shared ContentMost Shared
EurekaMag PDF Full Text ContentPDF Full Text
+ PDF Full Text
Request PDF Full TextRequest PDF Full Text
+ Follow Us
Follow on FacebookFollow on Facebook
Follow on TwitterFollow on Twitter
Follow on Google+Follow on Google+
Follow on LinkedInFollow on LinkedIn

+ Translate

Pneumocystis carinii antigen detection in rat serum and lung lavage

Journal of Clinical Microbiology 26(9): 1763-1771
Pneumocystis carinii antigen detection in rat serum and lung lavage
We developed a modified double-antibody sandwich enzyme-linked immunosorbent assay (ELISA) that detected relatively low concentrations of known Pneumocystis carinii antigen added to buffer or rat sera. Artificial immunization-derived polyclonal rabbit anti-P. carinii antibody was used on the solid phase to capture the antigen. Infection-derived (after P. carinii pneumonia) polyclonal rat anti-P. carinii antibody or a mixture of five murine monoclonal antibodies was used as the antigen detector antibody. Rabbit anti-rat immunoglobulin G antibody or goat anti-mouse immunoglobulin G antibody conjugated to alkaline phosphatase was used as the final antibody. After standardization and optimization of the various reactants in this ELISA system, approximately 53 ng of known P. carinii antigen per ml suspended in phosphate-buffered saline-Tween 20 buffer or 210 ng of antigen per ml suspended in normal rat serum diluted 1:4 could be detected. In addition, an indirect ELISA for P. carinii antibody measurement was developed, using as the antigen a soluble supernatant from a sonicated preparation of Percoll-purified whole cysts and trophozoites to coat the solid phase. Limited studies with sera from a small number of caesarian-obtained, barrier-sustained rats from Charles River Breeding Laboratories, Inc., and the National Institutes of Health and sera from normal and heavily infected rats indicated that the caesarian-obtained, barrier-sustained rats had negligible levels of antibody. The normal and heavily infected rats had variable antibody titers. A significantly high level of P. carinii antigenemia was detected in only 2 (11%) of 18 heavily infected rats. Extensive studies of the P. carinii pneumonia rat model with the ELISA did not reveal significant serum P. carinii antigenemia during the acute stage of infection. However, soluble P. carinii antigen was detected by the ELISA and Western blot assays in the supernatant of lavage fluid after centrifugation to sediment intact organisms. As expected, P. carinii antigens were detected by these assays in the lavage pellet recovered after centrifugation. In conclusion, the antigen assay used in this study detected P. carinii antigen in lung lavage but failed to detect P. carinii antigen in rat serum during the acute phase of infection.

(PDF 0-2 workdays service: $29.90)

Accession: 001912116

PMID: 3263387

Related references

Immunohistochemical detection of Pneumocystis carinii in transbronchial lung biopsy specimens: antigen difference between human and rat Pneumocystis carinii. Japanese Journal of Clinical Oncology 22(6): 387-392, 1992

Detection of antibodies to Pneumocystis carinii in bronchoalveolar lavage fluid by immunoreactivity to Pneumocystis carinii within alveoli, granulomas, and disseminated sites. Modern Pathology 5(2): 107-113, 1992

Detection of antibodies to pneumocystis carinii in bronchoalveolar lavage fluid bal in patients with and without pneumocystis carinii pneumonia pcp. Laboratory Investigation 64(1): 86A, 1991

Detection of pneumocystis carinii antigen in bronchoalveolar lavage bal by dot immunobinding assay dib. Sixth International Conference on Aids Sixth International Conference on Aids, Vols 1-3 Pagination Varies Sixth International Conference on Aids University Of California San Francisco: San Francisco, California, Usa Illus Maps Paper : abstract Thb 393, 1990

The detection of pneumocystis carinii in papanicolaou stained smears of lung lavage fluid sediment. American Review of Respiratory Disease 137(4 PART 2): 497, 1988

Diagnostic yield of bronchoalveolar lavage and bronchoscopic lung biopsy for detection of Pneumocystis carinii. Mayo Clinic Proceedings 71(11): 1025-1029, 1996

Identification and initial characterization of Pneumocystis carinii soluble antigens in rabbit serum and lung lavage. European Journal of Protistology 29(2): 246-253, 1993

The value of Pneumocystis carinii antibody and antigen detection for diagnosis of Pneumocystis carinii pneumonia after marrow transplantation. American Review of Respiratory Disease 120(6): 1283-1287, 1979

Detection of Pneumocystis carinii in rats by polymerase chain reaction: comparison of lung tissue and bronchoalveolar lavage specimens. Laboratory Animal Science 46(6): 628-634, 1996

Application of immunoblotting to detect soluble Pneumocystis carinii antigen(s) in bronchoalveolar lavage of patients with Pneumocystis pneumonia and AIDS. Journal of Clinical Pathology 43(7): 584-586, 1990