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3125 | LIGHT DIAGNOSTICS™ Respiratory Syncytial Virus DFA, ~50 tests

3125
2 mL  
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      Key ApplicationsFormatHostDetection Methods
      IF FITC M Fluorescent
      Description
      Catalogue Number3125
      Brand Family Chemicon®
      Trade Name
      • LIGHT DIAGNOSTICS
      • Chemicon
      DescriptionLIGHT DIAGNOSTICS™ Respiratory Syncytial Virus DFA, ~50 tests
      OverviewLight Diagnostics™ Respiratory Syncytial Virus DFA kit is intended for the detection and identification of respiratory syncytial virus (RSV) in inoculated cell cultures.

      For In Vitro Diagnostic Use.

      Test Principle:

      Light Diagnostics RSV DFA Kit utilizes a direct immunofluorescent antibody technique for identifying RSV in infected tissue cultures and cell preparations made from respiratory specimens. The monoclonal antibodies are labeled with FITC (fluorescein isothiocyanate), which fluoresce apple-green when illuminated with ultraviolet light. The labeled antibody will bind to viral antigen present in the specimen. Unbound reagent is removed by washing with buffer. Cells in positive specimens will fluoresce apple-green while uninfected cells will stain dull red due to the presence of Evans blue.

      Summary and Explanation:

      By age 2, most children have experienced respiratory syncytial virus (RSV) infection, making it the most important viral cause of childhood lower respiratory tract illness1. RSV infection usually results in colds with profuse rhinorrhea, but 25-40% of infants 6 weeks to 6 months old with primary infections, will develop lower respiratory tract illness1 and is responsible for more pneumonia and bronchiolitis than all other microbial pathogens2. Childhood RSV pneumonia and bronchiolitis may result in long-term respiratory abnormalities such as abnormal pulmonary function, asthma, and recurrent cough and bronchitis3.

      RSV has also been implicated in sudden infant death syndrome (SIDS) although the nature of the association is undetermined2.

      RSV belongs to the family Paramyxoviridae and the genus Pneumovirus. It is an enveloped pleomorphic virus ranging from 150-300 nm in diameter4,5,6 with a single-stranded RNA genome1.

      RSV may be detected by immunofluorescence, culture isolation and confirmation, EIA, complement fixation, or neutralization7,8,9. A variety of cell lines are suitable for RSV cultivation. For primary isolation, Hep-210 or HeLa5 are acceptable cell lines although others such as MRC-5, Vero, LLC-MK2, or CV-1 have been used. The virus produces characteristic cytopathic effects of syncytium formation and cell destruction2.
      Alternate Names
      • RSV DFA
      Materials Required but Not Delivered1. Cell culture for isolation of RSV. Each laboratory must maintain viable stocks of cells at appropriate passage-state that will efficiently allow replication of RSV from processed specimens. These cells must be checked periodically for ability to support growth of RSV. Appropriate cell lines can be obtained from the American Type Culture Collection (ATCC), 12 301 Parklawn Drive, Rockville, MD 20852.

      2.Viral transport medium (VTM), which is non-inhibitory to RSV and the tissue culture cells used for viral isolation containing Hank's Balanced Salt Solution (HBSS) with antibiotics and a protein stabilizer is a suitable medium. Avoid the use of animal sera (except precolostral fetal bovine serum) 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 fetal bovine serum (FBS)

      4. Sterile tissue culture tubes, dram vials, or multi-well plates

      5. Acetone, spectrophometric grade

      Note: Acetone is hygroscopic and should be kept in tightly stoppered bottles and stored at room temperature. 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 6-8mm in diameter

      7. Sterile pipettes

      8. Humid chamber

      9. Sodium hypochlorite solution (0.05%)

      10. No. 1 coverslips

      11. 37° incubator

      12. Sterile swabs

      13. Forceps

      14. Vials for collection and transport of specimens

      15. Fluorescence microscope with appropriate filter combination for FITC (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
      • RSV Reagent - (Catalog No. 5022). One 2 mL dropper vial containing FITC-labeled anti-RSV monoclonal antibodies in phosphate buffered saline (PBS), Tween 20, and 0.02% Evans blue, with sodium azide as a preservative.
      • RSV Control Slide - (Catalog No. 5012). Two control slides containing one RSV infected (positive) well and one non-infected (negative) well.
      • Phosphate Buffered Saline - (Catalog No. 5087) One packet of PBS salts to yield 1 liter upon reconstitution with distilled water. Store in a clean, closed container at room temperature.
      • Tween 20 / Sodium Azide Solution (100 X) - (Catalog No. 5037). One 10 mL Tween 20 and sodium azide concentrate to be diluted 1:100 in PBS.
      • Mounting Fluid - (Catalog No. 5013). One 10 mL dropper vial containing Tris buffer, glycerin, fluorescence enhancer, and sodium azide as preservative. Store at room temperature.
      Detection methodFluorescent
      FormatFITC
      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 2-8°C, the Respiratory Syncytial Virus DFA kit is stable up to the expiration date printed on the kit label. Do not freeze or expose to elevated temperatures. Discard any remaining reagents after the kit expiration date.

      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 conjugate 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:

      · Sodium azide (present in the conjugate, diluent, PBS solution, and mounting fluid) is toxic if ingested. Sodium azide 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 may cause erroneous results.

      · Avoid leaving reagents above 2-8°C for extended periods.

      · Acetone is extremely flammable and harmful if swallowed or inhaled. Keep away from heat, sparks or flame. Avoid breathing vapor. Use adequate ventilation.

      · Do not allow slides to dry at any time during the staining procedure.

      · Handle all specimens and materials as potentially infectious material. Decontaminate with 0.05% sodium hypochlorite (a l:100 dilution of household bleach).

      · Do not expose reagents to bright light during storage or incubation.

      · Avoid contact with Evans blue as it is a potential carcinogen. If skin contact occurs, flush with large volumes of water.

      · 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 of samples.
      Packaging Information
      Material Size2 mL
      Transport Information
      Supplemental Information
      Specifications

      Documentation

      MSDS

      Title

      Safety Data Sheet (SDS) 

      References

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      Darras S, Gerhart J, Terasaki M, Kirschner M, Lowe CJ
      Development (Cambridge, England) 138 959-70. 2011

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      21303849 21303849
      Improved detection of respiratory viruses in pediatric outpatients with acute respiratory illness by real-time PCR using nasopharyngeal flocked swabs
      Munywoki PK, Hamid F, Mutunga M, Welch S, Cane P, Nokes DJ
      Journal of clinical microbiology 49 3365-7. Epub 2011 Jul 20. 2011

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      Nitric oxide regulates Angiopoietin1/Tie2 expression after stroke.
      Alex Zacharek, Jieli Chen, Chunling Zhang, Xu Cui, Cynthia Roberts, Hao Jiang, Hua Teng, Michael Chopp
      Neuroscience letters 404 28-32 2006

      Pokaż streszczenie Pełny tekst artykułu
      16762501 16762501
      QuickVue influenza test for rapid detection of influenza A and B viruses in a pediatric population.
      Quach, Caroline, et al.
      Clin. Diagn. Lab. Immunol., 9: 925-6 (2002) 2002

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      12093698 12093698
      ApoA-I secretion by rabbit intestinal mucosa cell cultures.
      T L Carlson,B A Kottke
      Lipids 26 1991

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      1762512 1762512