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3303 | LIGHT DIAGNOSTICS™ Coxsackievirus B Blend Reagent, ~50 tests, included in kit #3350 & #3365

3303
2 mL  
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      Overview

      Replacement Information

      Key Spec Table

      Key ApplicationsFormatHostDetection Methods
      IFPurifiedMFluorescent
      Description
      Catalogue Number3303
      Brand Family Chemicon®
      Trade Name
      • LIGHT DIAGNOSTICS
      • Chemicon
      DescriptionLIGHT DIAGNOSTICS™ Coxsackievirus B Blend Reagent, ~50 tests, included in kit #3350 & #3365
      OverviewLight Diagnostics group specific monoclonal antibodies against coxsackie B viruses are intended for use in indirect fluorescence screening for the presumptive identification of coxsackie B1, B2, B3, B4, B5, and B6 viruses obtained in cell culture and not intended for testing directly on human specimens.

      Test Principle:

      Light Diagnostics Blend of Coxsackie B Viral Monoclonal Antibodies (MAb Cox B Blend) can be used to identify coxsackie B viral isolates in cell culture using an indirect immunofluorescence assay (IFA). The monoclonal antibodies provided will bind to the group specific enterovirus isolates present on the cell culture slide. 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. FITC exhibits an apple green fluorescence when illuminated by ultraviolet light allowing visualization of the complex by microscopy. A positive result is indicated by cell fluorescence. Non-infected cells stain a dull red if Evans blue counterstain is used in the FITC-labeled secondary antibody or used elsewhere in the procedure .

      Background and Clinical Significance:

      Enteroviruses are classified to be in the picornavirus family, pico [small] + RNA [ribonucleic acid] + virus. Picornaviruses are among the smallest and simplest ribonucleic acid containing viruses known [1]. The RNA for many enteroviruses have now been cloned and complete genomic sequences have been obtained. The RNA from all sequenced enteroviruses are similar in length, about 7400 nucleotides, and have identical organization [1].

      The human alimentary tract is the predominant site of enterovirus replication and these viruses were first isolated from enteric specimens. These viruses are the cause of paralytic poliomyelitis, aseptic meningitis-encephalitis, myocarditis, pleurodynia, hand-foot-and-mouth disease, conjunctivitis, and numerous other syndromes associated with extra-intestinal target organs. There are 67 numbered types of enteroviruses in the enteroviruses family [1]: polioviruses (3), coxsackieviruses A (23), coxsackieviruses B (6), echoviruses (31), and other enteroviruses (4)

      Enteroviruses, including echoviruses and coxsackieviruses, have been reported as the major etiologic agents of aseptic meningitis [2]. Clinical syndromes associated with infections by each type of enterovirus have also been reported [3]. Coxsackie B viruses can cause pleurodynia, infrequently paralysis, aseptic meningitis, severe systemic infection in infants, meningoencephalitis, pericarditis, myocarditis, upper respiratory illness, pneumonia, rash, hepatitis, and undifferentiated febrile illness.

      Establishing an association between an enterovirus and a particular disease in a patient requires laboratory confirmation of infection, usually by either isolation of the virus, or documentation of a specific serologic response in a properly timed specimen. Detailed descriptions of principals and procedures for diagnosis of enterovirus infections have been published [4-7]. Cell culture techniques have made the accurate detection of enteroviruses possible [8-10]. The identification of the enterovirus isolates will help prevention, treatment and understanding of the infectious diseases, and even discovery of new virus isolates. The typing of enterovirus isolates is generally accomplished by neutralization with type specific pools of immune sera [11]. This method is time consuming (7 days or more) and expensive. As an alternative, typing of enteroviruses with type specific monoclonal antibody and/or group specific monoclonal antibody pool(s) by the indirect immunofluorescence assay (IFA) is potentially more rapid and less expensive [12 - 18].
      Materials Required but Not Delivered· Acetone, reagent grade; stored in glass.

      · Distilled water.

      · Sodium hypochlorite solution, 0.05% (1:100 dilution of household bleach).

      · Sterile shell-vials with 12 mm coverslips containing monolayer of cell line appropriate for growth of enteroviruses.

      · Tissue culture media (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 enterovirus.

      · 0.1N NaOH.

      · 0.1N HCl.

      · Microscope slides, non-fluorescing.

      · No. 1 cover slips.

      · Negative and positive control slides.

      · Anti-Mouse IgG/FITC Conjugate (Catalog No. 5008).

      · Normal Mouse Antibody to be used as negative control.

      · Phosphate buffered saline (PBS, 0.01 M pH 7.1-7.4 with 0.085% NaCl and 0.1% azide), (Catalog No. 5087).

      · 0.05% Tween 20 /0.1% sodium azide solution (optional), (Catalog No. 5037).

      · Aspirator device with disposable sterile Pasteur pipettes.

      · 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.

      · Mounting Media (Catalog No. 5013).

      · Coxsackievirus B Control Slides (Catalog No. 5075).
      References
      Product Information
      Components
      • Blend of Coxsackievirus B Monoclonal Antibodies - (Catalog No. 3303).
      • One dropper vial containing 2 ml, sufficient for 25 tests, ready to use, mouse monoclonal antibodies against the coxsackie B viruses, protein stabilizer and 0.1 % sodium azide (preservative). (anti-B4 is a subclass IgG2b and the others are IgG2a.)
      Detection methodFluorescent
      FormatPurified
      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 monoclonal antibody is stable up to the expiration date printed on the label. Avoid multiple freeze and thaw.

      Warning and Precautions:

      *For in vitro diagnostic use.

      * The performance of Light Diagnostics coxsackievirus B Blend has not been determined on direct specimens.

      * Sodium azide, present in the reagents, can form potentially explosive metal azides with lead and copper pipes. As a precaution, flush with large amount of water to prevent azide build-up.

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

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

      * Handle all specimens and materials coming in contact with them as potentially infectious materials. All samples should be handled at the Biosafety Level 2 as recommended for any potentially infectious material in the Center for Disease Control/National Institute of Health Manual, "Biosafety in Microbiological and Biomedical Laboratories," (1984). Decontaminate with 0.05% sodium hypochlorite.

      * 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.
      Packaging Information
      Material Size2 mL
      Transport Information
      Supplemental Information
      Specifications

      Documentation

      References

      Reference overviewPub Med ID
      Neuronal Nogo-A modulates growth cone motility via Rho-GTP/LIMK1/cofilin in the unlesioned adult nervous system.
      Laura Montani,Bertran Gerrits,Peter Gehrig,Anissa Kempf,Leda Dimou,Bernd Wollscheid,Martin E Schwab
      The Journal of biological chemistry 284 2009

      Show Abstract Full Text Article
      19208621 19208621
      A homozygous COL6A2 intron mutation causes in-frame triple-helical deletion and nonsense-mediated mRNA decay in a patient with Ullrich congenital muscular dystrophy.
      Laura Lucarini, Betti Giusti, Rui-Zhu Zhang, Te-Cheng Pan, Cecilia Jimenez-Mallebrera, Eugenio Mercuri, Francesco Muntoni, Guglielmina Pepe, Mon-Li Chu
      Human genetics 117 460-6 2005

      Show Abstract
      16075202 16075202
      EMG and nerve conduction studies in children with congenital muscular dystrophy.
      Susana Quijano-Roy, Francis Renault, Norma Romero, Pascale Guicheney, Michel Fardeau, Brigitte Estournet
      Muscle nerve 29 292-9 2004

      Show Abstract
      14755496 14755496
      Ullrich scleroatonic muscular dystrophy is caused by recessive mutations in collagen type VI.
      O Camacho Vanegas, E Bertini, R Z Zhang, S Petrini, C Minosse, P Sabatelli, B Giusti, M L Chu, G Pepe
      Proceedings of the National Academy of Sciences of the United States of America 98 7516-21 2001

      Show Abstract Full Text Article
      11381124 11381124

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      Title
      LIGHT DIAGNOSTICS™ COXSACKIEVIRUS B BLEND MONOCLONAL ANTIBODIES