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3460 | LIGHT DIAGNOSTICS™ Pan-Enterovirus Detection Kit, ~125 tests

5 mL  
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      Key ApplicationsFormatHostDetection Methods
      IF Purified M Fluorescent
      Catalogue Number3460
      Trade Name
      • Chemicon
      DescriptionLIGHT DIAGNOSTICS™ Pan-Enterovirus Detection Kit, ~125 tests
      OverviewLight Diagnostics Pan-Enterovirus Detection Kit is intended as an indirect immunofluorescence screening assay for the preliminary identification of enteroviruses from cell culture and not intended for testing directly on human specimens.

      For in vitro diagnostic use.

      Test Principle:

      Light Diagnostics Pan-Enterovirus Blend can be used as a screening reagent for the preliminary identification of enteroviruses in cell culture using an indirect immunofluorescence assay (IFA). The monoclonal antibodies in the reagent will bind to enterovirus-infected cells and unbound monoclonal antibody is removed by rinsing with phosphate buffered saline (PBS). A secondary FITC (fluorescein isothiocyanate)-labeled antibody is then added which will bind to the antigen-antibody complex. Unbound secondary antibody is removed by rinsing with PBS and the resultant complex can be visualized by fluorescence microscopy. Uninfected cells stain a dull red if Evans blue counterstain is used in the FITC-labeled secondary antibody or elsewhere in the procedure.

      Summary and Explanation:

      Enteroviruses, classified as picornaviruses, pico [small] + RNA [ribonucleic acid] + virus, are among the smallest and simplest ribonucleic acid (RNA) containing viruses known1. The RNA from all sequenced enteroviruses are similar in length, about 7400 nucleotide, and have identical organization1.

      The human alimentary tract is the predominant site of enterovirus replication where they were first isolated from enteric specimens. There are 67 numbered types of enteroviruses in the enterovirus family1: three polioviruses, twenty-three coxsackieviruses A, six coxsackieviruses B, thirty-one echoviruses, and four other enteroviruses. These viruses cause a wide variety of disease ranging from paralytic poliomyelitis, aseptic meningitis-encephalitis, myocarditis, pleurodynia, hand-foot-and-mouth disease, conjunctivitis, and numerous other syndromes associated with extra-intestinal target organs.

      Enteroviruses, including echoviruses and coxsackieviruses, have been reported as major etiologic agents of aseptic meningitis2. Clinical syndromes associated with each type of enterovirus have also been reported3. Coxsackievirus A9 can cause aseptic meningitis, paralysis, exanthema, infant pneumonitis, and hepatitis; coxsackivirus A16 is a frequent cause of hand-foot-mouth disease. Coxsackieviruses B1-B6 can cause pleurodynia, aseptic meningitis, severe systemic infection in infants, meningoencephalitis, pericarditis, myocarditis, upper respiratory illness (cox B5) and undifferentiated febrile illness.

      Echoviruses can cause aseptic meningitis, paralysis, encephalitis, ataxia, Guillain-Barre' syndrome, exanthema and respiratory disease. They have also been associated with diarrhea, epidemic myalgia, pericarditis, myocarditis and hepatic disorders. Enteroviruses 70 and 71 can cause paralysis, meningoencephalitis, acute hemorrhagic conjunctivitis and hand-foot-and-mouth disease.

      Establishing an association between an enterovirus and a particular disease in a patient requires laboratory confirmation of infection by isolation of the virus or documentation of specific serologic response in a properly timed specimen. Detailed descriptions of principles and procedures for diagnosis of enterovirus infections have been published5-7. Cell culture techniques have made the accurate detection of enteroviruses possible8-12. The identification of the enterovirus isolates help in the prevention, treatment and understanding of the enterovirus illnesses, and even discovery of new virus isolates. The 'gold standard' for typing enterovirus isolates is neutralization with type-specific pools of immune sera12. This method is time consuming (7 days or more) and expensive. As an alternative, a presumptive identification of an enterovirus may be done by screening culture isolates with an immunofluorescent screening reagent. The isolate can then be further identified by the use of type-specific monoclonal antibodies and/or group-specific monoclonal antibody pool(s) by indirect immunofluorescence13-18.
      Materials Required but Not Delivered· Acetone, reagent grade; stored in glass
      · Distilled water or deionized water
      · Sodium hypochlorite solution, 0.05% (1:100 dilution of household bleach)
      · Sterile shell vials with 12 mm coverslips, or culture tubes containing monolayer of cell line appropriate for growth of enteroviruses (RMK, MRC-5, etc)
      · Tissue culture media such as RPMI or Eagle's Minimum Essential Medium (EMEM) with fetal bovine serum (FBS) and antibiotics, or equivalent
      · Viral transport medium which is non-inhibitory to enteroviruses
      · 0.1N NaOH
      · 0.1N HCl
      · Microscope slides, non-fluorescing
      · No. 1 cover slips
      · Negative and positive control slides
      · Mounting Fluid (Catalog No. 5013)
      · Anti-Mouse IgG:FITC Conjugate (Catalog No. 5008)
      · Normal Mouse Antibody (Catalog No. 5014)
      · PBS, 0.01 M, pH 7.1 to 7.4 with 0.085% NaCl and 0.1% sodium azide, (Catalog No. 5087)
      · 100X Wash Solution Concentrate (5% Tween 20 / 2% Sodium Azide Solution) (optional) (Catalog No. 5037)
      · Blocking solution, such as 1% casein or Difco Bacto Skim Milk with, 0.05% Tween 20, 0.02% sodium azide.
      · Centrifuge capable of 700-950 x g with biohazard buckets and adapters for shell vials
      · Fluorescence microscope with appropriate filter combination for FITC (excitation peak = 490 nm, emission peak = 520 nm) with 100x, 200x, 400x magnification (dry objective)
      · Forceps
      · Humid chamber
      · Incubator, 37 * 1*C
      · Syringe filter, 0.45 micron
      · Ultrasonic water bath
      · Vortex mixer or sonicator
      Product Information
      • Pan-Enterovirus Blend (Catalog No. 3360)
      • 1. One 5 mL dropper vial containing a blend of two mouse monoclonal antibodies which recognize all enteroviruses including echoviruses, coxsackie A and B viruses, polio 1,2, and 3, and enteroviruses 70 and 71, protein stabilizer and 0.1% sodium azide (preservative). The Pan-Enterovirus Blend may cross-react with hepatitis A, reovirus 3, astrovirus and rhinovirus.
      • 2. Pan-Enterovirus Control Slides - (Catalog No. 5069). Two control slides containing five infected (positive) wells and one non-infected (negative) wells. Each infected well represents a different enterovirus: Coxsackievirus A9, coxsackievirus B2, echovirus 30, enterovirus 71 and poliovirus 2. There is one uninfected (negative) well.
      • 3. Normal Mouse Antibody – (Catalog No. 5014). One 10 ml dropper vial containing normal mouse antibody to be used as a negative control.
      • 4. Anti-Mouse IgG:FITC – (Catalog No. 5008). One 10 ml dropper vials containing FITC labeled goat anti mouse IgG.
      • 5. Phosphate Buffered Saline - (Catalog No. 5087). One packet of Phosphate Buffered Saline Salts yields 1 liter when dissolved in distilled water. Store in a clean closed container at room temperature.
      • 6. Tween 20 / Sodium Azide Solution (100X) - (Catalog No. 5037). One 10 ml vial containing polyoxyethylene sorbitan monolaurate (Tween 20) and sodium azide (NaN3) concentrate to be diluted 1:100 in PBS.
      • 7. Mounting Fluid - (Catalog No. 5013). One 10 mL dropper vial containing Tris-buffered glycerin, a fluorescence enhancer, and 0.1% sodium azide (preservative).
      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
      • For in vitro Diagnostic Use
      • CE Mark
      Storage and Shipping Information
      Storage ConditionsWhen stored at 2-8°C, the Pan-Enterovirus Blend is stable up to the expiration date printed on the label. Do not freeze or expose to elevated temperatures. Discard any remaining reagent after the expiration date.

      Warning and Precautions:

      · Incubation times or temperatures other than those specified may give erroneous results. Any change must be validated by the user.

      * The performance of the Light Diagnostics Pan-Enterovirus Blend has not been determined on direct specimens.

      · The monoclonal antibodies present in the Pan-Enterovirus Detection Kit may show some background in certain cell lines, such as HEp-2 cells or some monkey cell lines.

      · Slides prepared too early (<25% CPE) or too late (>95% CPE) can be difficult to read and can lead to false negative results.

      · Sodium azide, present in the reagent, may react with lead and copper plumbing to form potentially explosive metal azides. Upon disposing of solutions that contain sodium azide, flush plumbing with a large volume of water to prevent build-up.

      * Handle all specimens and materials coming in contact with them as potentially infectious materials. Decontaminate with 0.05% sodium hypochlorite prior to disposal.

      * Avoid contact with Evans blue, if present in any reagent, as it is a potential carcinogen. If skin contact occurs, flush with large volumes of water.

      * Do not mouth pipette reagents.

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

      · Pooling or alteration of any reagent may cause erroneous results.

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

      · Mounting Fluid (Catalog No. 5013) 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 Size5 mL
      Transport Information
      Supplemental Information




      Scheda di sicurezza (MSDS) 

      Riferimenti bibliografici

      Panoramica dei riferimenti bibliograficiCodice d'identificazione nel Pub Med
      aPKC acts upstream of PAR-1b in both the establishment and maintenance of mammalian epithelial polarity.
      Atsushi Suzuki, Maki Hirata, Katsusi Kamimura, Rika Maniwa, Tomoyuki Yamanaka, Keiko Mizuno, Masaru Kishikawa, Hiroko Hirose, Yoshiko Amano, Natsuko Izumi, Yoshihiro Miwa, Shigeo Ohno
      Current biology : CB 14 1425-35 2004

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      Manuali d'uso

      LIGHT DIAGNOSTICS™ El Pan-Enterovirus Detection Kit - (French, Italian, Spanish & German Translation)
      LIGHT DIAGNOSTICS™ El Pan-Enterovirus Detection Kit - (French, Italian, Spanish & German Translation)