Comparison of histological and immunological techniques for detection of Pneumocystis carinii in rat bronchial lavage fluid

Milder, James E.; Walzer, Peter D.; Coonrod, J D; Rutledge, M E

doi:10.1128/jcm.11.4.409-417.1980

Cited by 39 publications

(10 citation statements)

References 30 publications

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“…Serologic diagnosis by the immunoblotting technique was quite effective in detecting antibodies indicative of exposure to P. carinii in all groups of rats prior to induction. The presence of antibodies to P. carinii antigens in nonimmunosuppressed rats has been reported extensively in the literature (4,7,20,31) but has not been routinely accepted as a criterion for the expected development of P. carinii pneumonia, since the presence of antibodies does not always correlate with development of pneumocystosis and their absence does not guarantee the lack of infection (7). Our study shows that Ͼ75% of a subpopulation of rats taken from an enzootically infected animal colony (representing about 5% of the male nonbreeding population) were seropositive to rat P. carinii.…”

Section: Discussionmentioning

confidence: 99%

Latent Pneumocystis carinii Infection in Commercial Rat Colonies: Comparison of Inductive Immunosuppressants plus Histopathology, PCR, and Serology as Detection Methods

Weisbroth¹,

Geistfeld

Weisbroth³

et al. 1999

J Clin Microbiol

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Histopathologic evaluation combined with a period of immunosuppression has been the standard procedure for detection ofPneumocystis carinii in commercial rat colonies. Variation in induction regimens and in the sensitivity of detection methods may result in underreporting of the presence of P. cariniiin breeding colonies or delay its detection. In the present study, methylprednisolone and cyclophosphamide were evaluated for the ability to induce P. carinii infection in rats from an enzootically infected commercial barrier colony. The presence ofP. carinii was detected by histopathologic methods and by amplification of a targeted region of the P. carinii thymidylate synthase gene by PCR over the 8-week study period. Sera taken from rats prior to either induction regimen were evaluated for the presence of P. carinii-specific antibodies by the immunoblotting technique. Few significant differences in ability to induce organism burden or in histopathology were observed between the two immunosuppressive regimens. However, a dramatic loss of weight over the study period was observed in rats treated with methylprednisolone but not in rats treated with cyclophosphamide. Although histopathologic changes attributable toP. carinii did not appear before 2 weeks with either immunosuppressant, the presence of the organism in these animals was detected by immunoblotting and PCR. Cyst scores and the intensities of the histopathologic lesions increased during the study period, but the number of rats exhibiting evidence of P. cariniiinfection did not change after week 3. These results suggest that use of the PCR method on postmortem lung tissue of rats without prior induction regimens or identification of anti-P. carinii antibodies in antemortem serum samples is a sufficiently sensitive method for detection of the presence of a P. cariniicarrier state in rodent breeding colonies.

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Section: Discussionmentioning

confidence: 99%

Latent Pneumocystis carinii Infection in Commercial Rat Colonies: Comparison of Inductive Immunosuppressants plus Histopathology, PCR, and Serology as Detection Methods

Weisbroth¹,

Geistfeld

Weisbroth³

et al. 1999

J Clin Microbiol

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“…Clinical and investigative interest in P. carinii has long focused on the morphological and staining properties of the organism. The most common type of study has involved staining of lung imprint smears or tissue sections to improve the rapidity of diagnosis (10,19,21,28). Stains (e.g., methenamine silver, toluidine blue 0, cresyl echt violet, and Gram Weigert) which selectively stain the wall of the cyst have been popular because they provide easy identification of this form of the organism (3,4,29).…”

Section: Discussionmentioning

confidence: 99%

Techniques for examining Pneumocystis carinii in fresh specimens

Ruffolo¹,

Cushion²,

Walzer³

1986

J Clin Microbiol

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Pneumocystis carinii was examined in fresh preparations of infected rat lung homogenates and tissue culture supernatants by a variety of light microscope techniques, vital dyes, and histologic stains. Phase-contrast microscopy, Nomarski interference-contrast microscopy, and bright-field microscopy with oblique illumination provided excellent views of P. carinii. Erythrosin B, and to a lesser extent trypan blue, were helpful in assessing organism viability. The use of Triton X-100-Giemsa stain permitted differentiation of the developmental stages in the P. carinii life cycle. The techniques developed here are easily adaptable to the microbiology laboratory and thus should have important clinical and research applications.

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“…Reagents used in the IFA technique were prepared according to procedures outlined in earlier studies (25,34,41).…”

Section: Methodsmentioning

confidence: 99%

Outbreaks of Pneumocystis carinii pneumonia in colonies of immunodeficient mice

et al. 1989

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Outbreaks of Pneumocystis carinii pneumonia occurred in colonies of nulnu and scidlscid mice at four different institutions. The disease, which was characterized by chronic wasting and respiratory insufficiency, was more severe in older mice and in animals housed in cages with special protective tops. Histopathologic features included alveolar filling with the typical foamy honeycomb material and a mild, nonspecific host inflammatory response. Immunofluorescence and immunoblotting studies suggested the P. carinni isolate was of mouse rather than of rat or human origin, and the outbreaks could be related to each other by common vendor or source of breeding animals. Once P. carinii became established in a mouse colony, the organism tended to persist for long periods of time. The principal control measure was depopulation of the colony, although limited experience with the administration of trimethoprim-sulfamethoxazole was encouraging. Thus, outbreaks of pneumocystosis are a serious problem among colonies of immunodeficient mice, with important implications for the use of these animals in biomedical research. Data obtained by studying these outbreaks should enhance understanding of the pathogenesis of P. carinii pneumonia and be helpful in formulating improved methods of detection and control.

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Comparison of histological and immunological techniques for detection of Pneumocystis carinii in rat bronchial lavage fluid

Cited by 39 publications

References 30 publications

Latent Pneumocystis carinii Infection in Commercial Rat Colonies: Comparison of Inductive Immunosuppressants plus Histopathology, PCR, and Serology as Detection Methods

Latent Pneumocystis carinii Infection in Commercial Rat Colonies: Comparison of Inductive Immunosuppressants plus Histopathology, PCR, and Serology as Detection Methods

Techniques for examining Pneumocystis carinii in fresh specimens

Outbreaks of Pneumocystis carinii pneumonia in colonies of immunodeficient mice

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