Pneumocystis Carinii

Pneumocystis pneumonia (PCP) is a form of pneumonia caused by the yeast-like fungal microorganism, Pneumocystis carinii. It is relatively rare in people with normal immune systems but common among people with AIDS. PCP can also develop in patients who are taking immunosuppressant medications (e.g., patients who have undergone solid organ transplantation) and in patients who have undergone bone marrow transplantation.

Pneumocystis carinii is detected in human tissues (typically in lungs of immunocompromised patients) in the trophozoite stage. Anti Pneumocystis carinii antibody reacts with an epitope on the organism that is resistant to formalin, picric acid, paraffin, as well as alcohol and xylene. No cross-reactivity has been demonstrated with other fungi or parasitic organisms.

Pneumocystis carinii is a mouse monoclonal antibody derived from cell culture supernatant that is concentrated, dialyzed, filter sterilized and diluted in buffer pH 7.5, containing BSA and sodium azide as a preservative.

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1. Cut and mount 3-4 micron formalin-fixed paraffin-embedded tissues on positive charged slides.
2. Air dry for 2 hours at 58° C.
3. Deparaffinize, dehydrate and rehydrate tissues.
4. Subject tissues to heat epitope retrieval using a suitable retrieval solution such as ImmunoDNA Retriever with Citrate (BSB 0020-BSB 0023) or EDTA (BSB 0030-BSB 0033).
5. Any of three heating methods may be used:
   • Electric Pressure Cooker
     Place standoff rack at base of pressure cooker. Add 1-2 inches of distilled water to the pressure cooker and turn heat to high, and incubate for 15 minutes. Open and immediately transfer slides to room temperature.
   • Water Bath Method
     Place tissues/slides in a pre-warmed staining dish or coplin jar containing the ImmunoDNA Retriever with Citrate or EDTA in a water bath set at 95°-99° C. Incubate for 30-60 minutes.
   • Conventional Steamer Method
     Place tissues/slides in a pre-warmed staining dish or coplin jar containing the ImmunoDNA Retriever with Citrate or EDTA in a Steamer, cover and steam for 30-60 minutes.
6. After heat treatment, transfer slides in ImmunoDNA Retriever with Citrate or EDTA to room temperature and let stand for 15-20 minutes.
7. Wash slides with IHC wash buffer or DI water.
8. Continue IHC staining protocol.