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3195 | LIGHT DIAGNOSTICS™ Pneumocystis carinii DFA Kit, ~50 tests (Replaces 3190)

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      Replacement Information

      Tabla espec. clave

      Key ApplicationsFormatHostDetection Methods
      IF FITC M Fluorescent
      Catalogue Number3195
      Brand Family Chemicon®
      Trade Name
      • Chemicon
      DescriptionLIGHT DIAGNOSTICS™ Pneumocystis carinii DFA Kit, ~50 tests (Replaces 3190)
      OverviewThe Light Diagnostics Pneumocystis carinii DFA kit is an in vitro procedure for the detection of Pneumocystis carinii cysts and trophozoites in direct smears of respiratory tract specimens including sputum, bronchoaveolar lavage, and lung biopsy.

      For in vitro diagnostic use.

      Test Principle:

      The Light Diagnostics Pneumocystis carinii DFA kit is a direct fluorescent antibody test for the detection of Pneumocystis carinii in direct smears of respiratory tract specimens including induced sputum, bronchoalveolar lavage (BAL), and lung biopsy specimens. The anti-Pneumocystis carinii monoclonal antibodies bind to both cysts and trophozoites of Pneumocystis carinii present in a respiratory tract specimen. These monoclonal antibodies are labeled with fluorescein isothiocyanate (FITC). FITC exhibits an apple-green fluorescence when illuminated by ultraviolet light allowing visualization of the complex by fluorescence microscopy. Observation of fluorescence and the characteristic morphology of Pneumocystis carinii cysts and trophozoites indicate the presence of Pneumocystis carinii and constitutes a positive result. If the specimen contains no Pneumocystis carinii cysts and trophozoites, no fluorescent immune complex is formed and the specimen is considered negative.

      Summary and Explanation:

      Pneumocystis carinii is an opportunistic pulmonary protozoan pathogen which is composed of two basic forms (6):

      a) The trophozoite is a small (1-4 micron) pleomorphic form which can be stained and identified in lung imprint smears.

      b) The cyst, which is relatively larger (5-7 micron), is less common. The cyst has a thick wall and contains up to 8 daughter forms, termed sporozoites. The cyst can be identified by stains that are selective for its cell wall.

      In addition, some intermediate forms and extracystic material can be observed when using the immunofluorescence method (14).

      Pneumocystis carinii causes interstitial plasma cell pneumonia and occurs in malnourished, debilitated, premature infants and children with primary immunodeficiency diseases. It develops in patients of all ages receiving immunosuppressive agents for treatment of cancer, organ transplants, and other disorders (1-4). Recently, Pneumocystis carinii has emerged as the most frequent opportunistic infection in patients with acquired immunodeficiency syndrome (AIDS). Pneumocystis carinii pneumonia (PCP) accounts for 43% to 85% of pulmonary infection associated with AIDS (4,14). Typical symptoms include prolonged breathlessness, dry cough, fever, and an inability to take a deep breath.

      The presenting manifestation of PCP can be subtle and non-specific. Rapid early detection of PCP improves prognosis through early therapeutic intervention. Pneumocystis carinii has been detected by methenamine silver, Giemsa, Gram-Weigert, and toluidine blue O stains (7-13). The organism has also been identified with indirect fluorescent monoclonal antibody (14,15).

      The Light Diagnostics Pneumocystis carinii DFA kit utilized directly conjugated fluorescent antibodies to provide clear, easy to interpret results in only 15 minutes.
      Materials Required but Not Delivered1. Control slides: Positive and negative control slides prepared from previously tested, known positive and negative patient specimens may be used. Control positive (Cat. No. 2416010) and negative (Cat No. 2419010) slides may also be purchased from Remel titled PC Slides, positive and negative Pneumocystic for FA.

      2. Microscope Slides: PTFE-coated, each containing 2 wells.

      3. Specimen collection devices and solutions for bronchoalveolar lavage (BAL).

      4. Specimen collection devices for induced sputum, nebulizer, 3-10% saline solution.

      5. Cytospin centrifuge, cones, slides, filters, and slide holders for preparation of cytospins (Shandon Southern, Sewickley, PA).

      6. Mucolytic/sputum clearing reagent.

      7. Bovine serum albumin, 22% solution.

      8. Devices for obtaining lung biopsy and for preparation of lung imprint smears, stomacher, stainless steel sieves or gauze.

      9. Test tubes and racks.

      10. Pipettes and / or micropipettes and safety pipeting devices.

      11. Coverslips.

      12. Methanol.

      13. Humid chamber.

      14. Gloves.

      15. Laminar flow hood, hair dryer, or small fan (optional).

      16. Centrifuge.

      17. Coplin jars (or equivalent).

      18. Lint-free tissues or cotton swabs.

      19. Applicator sticks.

      20. Fluorescence microscope equipped with the manufacturer recommended -filter system for fluorescein isothiocyanate: maximum excitation -wavelength 490 nm and mean emission wavelength 520 nm.

      21. Timer or stop watch.

      22. Absolute Ethanol.
      Product Information
      • PCP DFA Reagent - (Catalog No. 5195). One (1) 2 mL dropper vial of a mixture of two monoclonal antibodies against Pneumocystis carinii directly conjugated to FITC in a solution containing counterstain, stabilizers, and 0.1% sodium azide (NaN3).
      • PBS Buffer Salts - (Catalog No. 5087). One (1) packet of phosphate-buffered saline salts yield 1 liter when dissolved in distilled water. Store in a clean closed container at room temperature.
      • Tween 20 / Sodium Azide Solution (100x) - (Catalog No. 5037). One (1) 10 mL vial of polyoxyethylene sorbitan monolaurate (Tween 20) and sodium azide (NaN3) concentrate to be diluted 1:100 in PBS.
      • Mounting Medium - (Catalog No. 5013). One (1) 10 mL dropper vial containing tris buffer, glycerin, fluorescence enhancer, and sodium azide as preservative. Store at room temperature.
      Detection methodFluorescent
      Key Applications
      • Immunofluorescence
      Biological Information
      Antibody TypeMonoclonal Antibody
      Physicochemical Information
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Usage Statement
      • CE Mark
      • For in vitro Diagnostic Use
      Storage and Shipping Information
      Storage ConditionsKit components should be stored protected from light at 2-8°C, until the expiration date of the kit. Kit components must be brought to room temperature prior to use.

      During incubation, slides should be protected from light and kept in a humid chamber at the recommended temperature.

      A marked decrease in fluorescence may indicate antibody deterioration. A positive control should be tested with each specimen to ensure proper functioning of these reagents and proper staining procedure. If after appropriate analysis there is a decrease in staining intensity, discontinue use of the reagents.

      Warnings and Precautions:

      · Do not mix components from different kit lots.

      · Do not use kit components beyond expiration date.

      · Certain reagents in the Light Diagnostics Pneumocystis carinii DFA kit contain sodium azide which may react with lead or copper plumbing to form potentially explosive metal azides. When disposing of such reagents, always flush with copious amounts of water to prevent build up.

      · Patient specimens can contain infectious agents, especially those obtained from AIDS patients. All specimens should be handled with extreme caution and should be regarded as potentially infectious.

      · Gloves should be worn throughout the acetone fixation step and preferably throughout the entire procedure. Wash hands thoroughly after completion of test procedure.

      · Do not mouth pipette.

      · Avoid exposing the fluorescent antibody reagent to bright light during storage or incubation.

      · Avoid scratching the fixed specimens on the surface of the slide when adding reagents to the slide wells.

      · Do not allow the microscope slide to dry at any time during the staining procedure.
      Packaging Information
      Material Size2 mL
      Transport Information
      Supplemental Information


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      Referencias bibliográficas

      Visión general referenciasPub Med ID
      Trehalose-recycling ABC transporter LpqY-SugA-SugB-SugC is essential for virulence of Mycobacterium tuberculosis.
      Rainer Kalscheuer,Brian Weinrick,Usha Veeraraghavan,Gurdyal S Besra,William R Jacobs
      Proceedings of the National Academy of Sciences of the United States of America 107 2010

      Mostrar resumen Artículo Texto completo
      21118978 21118978
      Development and validation of an HPLC method for the determination of penicillin antibiotics residues in bovine muscle according to the European Union Decision 2002/657/EC.
      Victoria F Samanidou,Styliani A Nisyriou,Ioannis N Papadoyannis
      Journal of separation science 30 2007

      Mostrar resumen
      17960837 17960837

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