CMV Brite™ Turbo Kit

Introduction:
The CMV pp65 antigenemia assay is a valuable tool in the diagnosis and monitoring of active CMV infection in solid organ and bone marrow transplant patients as well as in the diagnosis and monitoring of CMV disease in AIDS patients. Early and rapid diagnosis of CMV infection is of great importance in avoiding over treatment with immunosuppressive drugs and in guiding antiviral therapy.

The CMV Brite™ Turbo antigenemia kit uses the well defined C10/C11 antibody cocktail to detect the CMV lower matrix phosphoprotein (pp65), an early antigen in virus replication, which is abundantly present in antigen-positive polymorphonuclear cells. The CMV Brite™ Turbo Kit, is a rapid new version of the first FDA registered immunofluorescence antigenemia kit for in vitro CMV diagnosis (1).

Special features of the CMV Brite™ Turbo Kit

  • A complete kit containing all the reagents needed from cell preparation to immunofluorescence staining.
  • A cocktail of antibodies specific for pp65. Antibodies C10/C11 are highly specific for pp65 and have been well documented (2,3,4,5).
  • Detection of CMV pp65 positive PMNs by indirect immunofluorescence gives high sensitivity and easy reading of the results.
  • The number of CMV pp65 positive cells are counted per duplicate stain.
  • Control slides are included. Each slide contains one negative and one positive control spot. The positive control spot is a combination of normal blood leukocytes and pp65 transfected cells.
  • Shelf life of at least 18 months.
  • Completed within 2 hours of sample collection
  • Studies for registration with FDA (USA), PEI (Germany), ADM (France) for use as in vitro diagnostic assay in progress.

Materials and reagents supplied in the CMV Brite™ Turbo Kit.

  • Erythrocyte Lysing Solution
  • Fixative solution
  • Permeabilization solution
  • Mouse monoclonal antibody C10/C11 specific for CMV pp65
  • FITC conjugated sheep anti-mouse immunoglobulin with Evans Blue
  • Control slides. Each slide containing one negative and one CMV pp65 positive cytospot

Sensitivity and Specificity
Patient studies in 3 different clinical institutions show that the CMV Brite™ Turbo Kit has the same high sensitivity and specificity as the CMV Brite™ Kit (1). There is no evidence of cross reactivity between other viruses and CMV using the CMV antigenemia assay using the C10/C11 monoclonal antibody coctail.

Results using CMV Brite™ Turbo Kit
Figure 2 Leukocytes stained with the CMV Brite™ Turbo antigenemia Kit. Immunofluorescence staining of a CMV pp65 antigen positive PMN. Negative cells are stained red with Evans Blue.



References
1. Landry,M.L. et al. Evaluation of CMV Brite Kit for detection of Cytomegalovirus pp65 antigenemia in peripheral blood leucocytes by immunofluroescence. J.Clin.Microbiol. 34. 1337-1339
2. Van Son, W.J., The,T.H., (1989) Cytomegaovirus infection after organ transplantation: an update with special emphasis on renal transplantation. Transpl. Int. 2, 147-164.
3. The, T.H., et al, (1990) Cytomegalovirus antigenemia. Rev. Inf. Dis. 12., S737-S744.
4. Van der Bij et al., (1988) Comparison between viremia and antigenemia for detection of cytomegalovirus in blood. J. Clin. Microbiology 26. 2531-2535
5. Grefte, J.M.M et al (1992) The lower matrix protein pp65 is the principal viral antigen present in peripheral blood leukocytes during an active cytomegalovirus infection. J.Gen. Virol. 73. 2923-2932
For research use only. Not for use in humans. For in vitro use only.




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