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3297 | LIGHT DIAGNOSTICS™ SimulFluor® Para 1,2/3, ~50 tests

3297
2 mL  
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      Overview

      Replacement Information

      Key Spec Table

      Key ApplicationsFormatHostDetection Methods
      IFSimulFluorMFluorescent
      Description
      Catalogue Number3297
      Brand Family Chemicon®
      Trade Name
      • LIGHT DIAGNOSTICS
      • SimulFluor
      • Chemicon
      DescriptionLIGHT DIAGNOSTICS™ SimulFluor® Para 1,2/3, ~50 tests
      OverviewIntended Use
      Light Diagnostics™ SimulFluor® Parainfluenza 1, 2/3 Immunofluorescence Assay is intended for the simultaneous detection of parainfluenza 1 and 2, and identification of parainfluenza 3 following amplification in cell culture.
      For In Vitro Diagnostic Use.

      Summary and Explanation
      Respiratory viruses are responsible for a significant proportion of illness in human populations. Some viruses (e.g. influenza) are seasonal while others (e.g. adenovirus) predominantly affect different age groups. In a given population, respiratory viruses may be responsible for a considerable amount of morbidity. The advent of appropriate anti-viral therapy against some respiratory viruses makes rapid screening to identify these organisms imperative, allowing early institution of therapy. The two most widely used anti-viral agents today are amantadine/rimantadine for influenza A and ribavirin for respiratory syncytial virus bronchiolitis. Both agents have varying modes of action although most involve early infection steps such as penetration or uncoating (12,19). At physiologically attainable concentrations, amantadine and rimantadine specifically inhibit replication of influenza A (4). Ribavirin has broad spectrum activity in vitro against a host of viruses such as respiratory syncytial virus, measles, influenza A and B, and parainfluenza (7,13,14).
      Parainfluenza viruses, combined with respiratory syncytial virus, represent the most significant upper respiratory pathogens in infants and young children (1,2,3,5). Four types of parainfluenza virus have been identified in children and adults. Types 1 and 2 are major causes of laryngotracheo-bronchitis (croup). The severity of illness is greatest in children ages 2 to 4 years (6). Parainfluenza type 3 infection can also lead to croup and is second only to respiratory syncytial virus as a cause of infant bronchiolitis and pneumonia (8,9,10,11,15). Illness from type 3 infection is most severe in infants less than 1 year old (6). In older children and adults, illness may be asymptomatic or mimic the common cold (16). Severe croup in early childhood or infancy may lead to bronchial hyperactivity in older children or adolescents after exercise. However, it remains undetermined whether bronchial hyperactivity was a pre-existing condition, which played a role in the pathogenesis of croup or whether it developed as a complication of severe croup illness (17,18).
      Parainfluenza type 4 has been associated only with mild upper respiratory illness in adults and children and is difficult to identify in cell culture (6).
      Parainfluenza viruses belong to the genus Paramyxovirus of the family Paramyxoviridae. They are enveloped viruses with a single-strand RNA genome of negative polarity and range in diameter from 150 to 200 nm (20).
      Parainfluenza viruses grow well in primary simian or human kidney cell lines and in LLC-¬MK2, a rhesus kidney heteroploid cell line (21). Trypsin is needed in the medium for the recovery of types 1 and 2, but not 3. Virus infection of tissue culture can be recognized by hemadsorption of guinea pig erythrocytes. Types 2 and 3 can be recognized by syncytium formation.
      The most sensitive method for identifying parainfluenza viruses is culture isolation and confirmation; it is the standard method for most laboratories. Light Diagnostics™ SimulFluor® Parainfluenza 1, 2/ 3 Immunofluorescence Assay utilizes monoclonal antibodies to simultaneously detect parainfluenza 1 and 2, and identify parainfluenza 3 following amplification in culture to provide clear, easy-to-interpret results.

      Test Principle

      Light Diagnostics™ SimulFluor® Parainfluenza 1, 2/ 3 Immunofluorescence Assay utilizes a single reagent for the one step simultaneous detection of parainfluenza 1 and 2, and identification of parainfluenza 3 in infected cell culture. The Light Diagnostics™ SimulFluor® Para 1, 2/ 3 Reagent contains two components. The primary component is directed against parainfluenza 1 and 2; the second component is specifically directed against parainfluenza 3. The monoclonal antibodies in the components will bind to the appropriate viral antigen on the specimen slide. Unbound antibody is washed from the slide with phosphate buffered saline (PBS). Illumination with ultraviolet light allows visualization of the antigen-antibody complexes by fluorescence microscopy. Parainfluenza 1 and 2 infected cells will exhibit an apple-green fluorescence; while, parainfluenza 3 infected cells will exhibit a yellow-gold fluorescence. The primary component stains the parainfluenza 1 and 2 infected cells a bright apple-green but does not differentiate between these viruses. The secondary component stains the parainfluenza 3 infected cells bright yellow-gold and allows for the differentiation of parainfluenza 3 from parainfluenza 1 and 2 in a single well. Uninfected cells stain dull red due to the presence of Evans blue counterstain in the reagent.
      Materials Required but Not Delivered1. Cell culture for isolation of respiratory viruses: Each laboratory must maintain viable stocks of cells at appropriate passage state that will efficiently allow replication of respiratory viruses from processed patient specimens. These cells must be checked periodically for ability to support growth of respiratory viruses. Appropriate cell lines can be obtained from the American Type Culture Collection (ATCC), 12301 Parklawn Drive, Rockville, MD 20852.
      2. Viral transport medium (VTM), which is non-inhibitory to the respiratory viruses and the tissue culture cells used for viral isolation: Hank's balanced salt solution (HBSS) with antibiotics and a protein stabilizer is a suitable medium. Avoid use of animal sera (except precolostral fetal bovine serum (FBS)) as protein stabilizer to prevent interference from inherent antibody.
      3. Tissue culture media such as RPMI or Eagle's Minimum Essential Medium (EMEM) with appropriate amount of precolostral FBS can be used.
      4. Sterile tissue culture tubes, dram vials, or multi well plates
      5. Acetone, reagent grade or better
      Note: Acetone is hygroscopic and should be kept in tightly stoppered bottles. Presence of moisture in the acetone may result in a hazy appearance on the substrate during fluorescence microscopy.
      6. Acetone-cleaned glass slides with wells at least 6mm in diameter in a hydrophobic mask
      7. Sterile pipettes
      8. Humid chamber
      9. Sodium hypochlorite solution (0.05%)
      10. No. 1 coverslips
      11. 37°C incubator with rheostat for temperature regulation
      12. Sterile swabs
      13. Forceps
      14. Vials for collection and transportation of specimens
      15. Fluorescence microscope with appropriate filter combination for fluorescein (excitation peak 490 nm, emission peak 520 nm)
      16. Sterile glass beads (1 to 3 mm diameter)
      17. Centrifuge
      18. Vortex mixer or sonicator
      19. Distilled or deionized water
      References
      Product Information
      Components
      • SimulFluor® Para 1, 2/ 3 Reagent - (Catalog No. 5297).
      • Para 1, 2 ,3 Control Slide - (Catalog No. 5011).
      • Phosphate Buffered Saline - (Catalog No. 5087).
      • Tween 20 / Sodium Azide Solution (100X) - (Catalog No. 5037).
      • Mounting Fluid - (Catalog No. 5013).
      Detection methodFluorescent
      FormatSimulFluor
      Applications
      Key Applications
      • Immunofluorescence
      Biological Information
      HostMouse
      Antibody TypeMonoclonal Antibody
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Usage Statement
      • For in vitro Diagnostic Use
      • CE Mark
      Storage and Shipping Information
      Storage ConditionsWhen stored at 2° to 8°C, the Light Diagnostics™ SimulFluor® Para 1, 2/ 3 Reagent is stable up to the expiration date printed on the kit label. Do not freeze or expose to elevated temperatures. Discard any remaining reagent after the kit expiration date.

      Warnings and Precautions

      • Sodium azide (present in the conjugate, monoclonal antibodies, wash buffer, and mounting fluid) can react with lead or copper plumbing to form potentially explosive metal azides. When disposing of these materials, flush with large volumes of water to prevent azide build-up.
      • Pooling or diluting conjugates or monoclonal antibodies may cause erroneous results.
      • Do not allow slides to dry at any time during the staining procedure.
      • Handle all specimens and materials as if potentially infectious. Decontaminate with 0.05% sodium hypochlorite (a 1:100 dilution of household bleach) prior to disposal.
      • Do not expose reagents to bright light during storage or incubation.
      • Avoid contact with Evans blue (present in the Light Diagnostics™ SimulFluor® Para 1, 2/ 3 Reagent), as it is a potential carcinogen. If skin contact occurs, flush with large volumes of water.
      • Acetone is extremely flammable and harmful if swallowed or inhaled. Keep away from heat, sparks, or flames. Avoid breathing vapor. Use adequate ventilation.
      • Do not mouth pipette reagents.
      • Do not substitute reagents from other manufacturers.
      • Alteration of protocol provided may cause erroneous results.
      • When staining multiple samples on a slide, avoid cross contamination between samples.
      • Mounting Fluid contains a fluorescence enhancer that may be destructive to mucous membranes. Avoid direct skin or mucous membrane contact. If contact occurs, flush with large volumes of water.
      Packaging Information
      Material Size2 mL
      Transport Information
      Supplemental Information
      Specifications

      Documentation

      SDS

      Title

      Safety Data Sheet (SDS) 

      References

      Reference overviewPub Med ID
      Fate of influenza A virion proteins after entry into subcellular fractions of LLC cells and the effect of amantadine
      Richman, D D, et al
      Virology, 151:200-10 (1986) 1986

      3705464 3705464
      Ribavirin aerosol treatment of influenza B virus infection.
      McClung, H W, et al.
      JAMA, 249: 2671-4 (1983) 1983

      Show Abstract
      6341642 6341642
      Pulmonary function and bronchial reactivity in children after croup
      Gurwitz, D, et al
      Am Rev Respir Dis, 122:95-9 (1980) 1980

      7406345 7406345
      Inhibition of influenza virus uncoating by rimantadine hydrochloride
      Koff, W C and Knight, V
      J Virol, 31:261-3 (1979) 1979

      501798 501798
      Use of immunoperoxidase for the rapid identification of human myxoviruses and paramyxoviruses in tissue culture.
      Benjamin, D R and Ray, C G
      Applied microbiology, 28: 47-51 (1974) 1974

      Show Abstract
      4367457 4367457
      Epidemiology of respiratory syncytial virus infection in Washington, D.C. I. Importance of the virus in different respiratory tract disease syndromes and temporal distribution of infection
      Kim, H W, et al
      Am J Epidemiol, 98:216-25 (1973) 1973

      4355005 4355005
      Parainfluenza pneumonia in adults
      Wenzel, R P, et al
      JAMA, 221:294-5 (1972) 1972

      4338336 4338336
      Broad-spectrum antiviral activity of Virazole: 1-beta-D-ribofuranosyl-1,2,4-triazole-3-carboxamide
      Sidwell, R W, et al
      Science, 177:705-6 (1972) 1972

      4340949 4340949
      Respiratory syncytial virus. I. Virus recovery and other observations during 1960 outbreak of bronchiolitis, pneumonia, and minor respiratory diseases in children
      CHANOCK, R M, et al
      JAMA, 176:647-53 (1961) 1961

      13692354 13692354
      Clinical features of infection with hemadsorption viruses
      PARROTT, R H, et al
      N Engl J Med, 260:731-8 (1959) 1959

      13644577 13644577

      Brochure

      Title
      Light Diagnostics™ products for infectious disease

      User Guides

      Title
      LIGHT DIAGNOSTICS™ SimulFluor® PARAINFLUENZA 1, 2 /3 DFA KIT