Skip to main content
Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1997 Feb;35(2):393–397. doi: 10.1128/jcm.35.2.393-397.1997

A novel Western blot test containing both viral and recombinant proteins for anticytomegalovirus immunoglobulin M detection.

T Lazzarotto 1, G T Maine 1, P Dal Monte 1, A Ripalti 1, M P Landini 1
PMCID: PMC229587  PMID: 9003603

Abstract

We devised a novel Western blot (WB) test for anti-human cytomegalovirus (HCMV) immunoglobulin M (IgM) detection which contains viral structural polypeptides, significant portions of recombinant p150 (ppUL32), and a significant portion of the most immunogenic nonstructural protein p52 (ppUL44). This new test was evaluated in latently infected blood donors, pregnant women, and transplant recipients with ongoing HCMV infection and shown to be more sensitive and specific than traditional WB and conventional enzyme immunoassay for the detection of HCMV-specific IgM.

Full Text

The Full Text of this article is available as a PDF (206.7 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Basson J., Tardy J. C., Aymard M. Pattern of anti-cytomegalovirus IgM antibodies determined by immunoblotting. A study of kidney graft recipients developing a primary or recurrent CMV infection. Arch Virol. 1989;108(3-4):259–270. doi: 10.1007/BF01310938. [DOI] [PubMed] [Google Scholar]
  2. Boland G. J., de Gast G. C., Hené R. J., Jambroes G., Donckerwolcke R., The T. H., Mudde G. C. Early detection of active cytomegalovirus (CMV) infection after heart and kidney transplantation by testing for immediate early antigenemia and influence of cellular immunity on the occurrence of CMV infection. J Clin Microbiol. 1990 Sep;28(9):2069–2075. doi: 10.1128/jcm.28.9.2069-2075.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Chee M. S., Bankier A. T., Beck S., Bohni R., Brown C. M., Cerny R., Horsnell T., Hutchison C. A., 3rd, Kouzarides T., Martignetti J. A. Analysis of the protein-coding content of the sequence of human cytomegalovirus strain AD169. Curr Top Microbiol Immunol. 1990;154:125–169. doi: 10.1007/978-3-642-74980-3_6. [DOI] [PubMed] [Google Scholar]
  4. Dolan J., Briggs J. D., Clements G. B. Antibodies to cytomegalovirus in renal allograft recipients: correlation with isolation of virus. J Clin Pathol. 1989 Oct;42(10):1070–1077. doi: 10.1136/jcp.42.10.1070. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Gleaves C. A., Smith T. F., Shuster E. A., Pearson G. R. Rapid detection of cytomegalovirus in MRC-5 cells inoculated with urine specimens by using low-speed centrifugation and monoclonal antibody to an early antigen. J Clin Microbiol. 1984 Jun;19(6):917–919. doi: 10.1128/jcm.19.6.917-919.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Gold D., Ashley R., Handsfield H. H., Verdon M., Leach L., Mills J., Drew L., Corey L. Immunoblot analysis of the humoral immune response in primary cytomegalovirus infection. J Infect Dis. 1988 Feb;157(2):319–326. doi: 10.1093/infdis/157.2.319. [DOI] [PubMed] [Google Scholar]
  7. Grazia Revello M., Zavattoni M., Percivalle E., Grossi P., Gerna G. Correlation between immunofluorescent detection of human cytomegalovirus immediate early antigens in polymorphonuclear leukocytes and viremia. J Infect Dis. 1989 Jul;160(1):159–160. doi: 10.1093/infdis/160.1.159. [DOI] [PubMed] [Google Scholar]
  8. Jiwa N. M., Van Gemert G. W., Raap A. K., Van de Rijke F. M., Mulder A., Lens P. F., Salimans M. M., Zwaan F. E., Van Dorp W., Van der Ploeg M. Rapid detection of human cytomegalovirus DNA in peripheral blood leukocytes of viremic transplant recipients by the polymerase chain reaction. Transplantation. 1989 Jul;48(1):72–76. doi: 10.1097/00007890-198907000-00017. [DOI] [PubMed] [Google Scholar]
  9. Kraat Y. J., Stals F. S., Christiaans M. H., Lazzarotto T., Landini M. P., Bruggeman C. A. IgM antibody detection of ppUL80A and ppUL32 by immunoblotting: an early parameter for recurrent cytomegalovirus infection in renal transplant recipients. J Med Virol. 1996 Mar;48(3):289–294. doi: 10.1002/(SICI)1096-9071(199603)48:3<289::AID-JMV13>3.0.CO;2-8. [DOI] [PubMed] [Google Scholar]
  10. Landini M. P., Lazzarotto T., Maine G. T., Ripalti A., Flanders R. Recombinant mono- and polyantigens to detect cytomegalovirus-specific immunoglobulin M in human sera by enzyme immunoassay. J Clin Microbiol. 1995 Oct;33(10):2535–2542. doi: 10.1128/jcm.33.10.2535-2542.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Landini M. P., Re M. C., Mirolo G., Baldassarri B., La Placa M. Human immune response to cytomegalovirus structural polypeptides studied by immunoblotting. J Med Virol. 1985 Dec;17(4):303–311. doi: 10.1002/jmv.1890170403. [DOI] [PubMed] [Google Scholar]
  12. Landini M. P., Ripalti A. A DNA-nicking activity associated with the nucleocapsid of human cytomegalovirus. Arch Virol. 1982;73(3-4):351–356. doi: 10.1007/BF01318089. [DOI] [PubMed] [Google Scholar]
  13. Landini M. P., Rossier E., Schmitz H. Antibodies to human cytomegalovirus structural polypeptides during primary infection. J Virol Methods. 1988 Dec;22(2-3):309–317. doi: 10.1016/0166-0934(88)90113-9. [DOI] [PubMed] [Google Scholar]
  14. Lazzarotto T., Dalla Casa B., Campisi B., Landini M. P. Enzyme-linked immunoadsorbent assay for the detection of cytomegalovirus-IgM: comparison between eight commercial kits, immunofluorescence, and immunoblotting. J Clin Lab Anal. 1992;6(4):216–218. doi: 10.1002/jcla.1860060409. [DOI] [PubMed] [Google Scholar]
  15. Lazzarotto T., Furlini G., Re M. C., Ramazzotti E., Campisi B., Landini M. P. Human cytomegalovirus replication correlates with differentiation in a hematopoietic progenitor cell line and can be modulated by HIV-1. Arch Virol. 1994;135(1-2):13–28. doi: 10.1007/BF01309762. [DOI] [PubMed] [Google Scholar]
  16. Lazzarotto T., Maine G. T., Del Monte P., Frush H., Shi K., Landini M. P. Detection of serum immunoglobulin M to human cytomegalovirus by western blotting correlates better with virological data than detection by conventional enzyme immunoassay. Clin Diagn Lab Immunol. 1996 Sep;3(5):597–600. doi: 10.1128/cdli.3.5.597-600.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Maine G. T., Lazzarotto T., Chovan L. E., Flanders R., Landini M. P. The DNA-binding protein pUL57 of human cytomegalovirus: comparison of specific immunoglobulin M (IgM) reactivity with IgM reactivity to other major target antigens. Clin Diagn Lab Immunol. 1996 May;3(3):358–360. doi: 10.1128/cdli.3.3.358-360.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Marsano L., Perrillo R. P., Flye M. W., Hanto D. W., Spitzer E. D., Thomas J. R., Murray P. R., Windus D. W., Brunt E. M., Storch G. A. Comparison of culture and serology for the diagnosis of cytomegalovirus infection in kidney and liver transplant recipients. J Infect Dis. 1990 Mar;161(3):454–461. doi: 10.1093/infdis/161.3.454. [DOI] [PubMed] [Google Scholar]
  19. Nielsen C. M., Hansen K., Andersen H. M., Gerstoft J., Vestergaard B. F. An enzyme labelled nuclear antigen immunoassay for detection of cytomegalovirus IgM antibodies in human serum: specific and non-specific reactions. J Med Virol. 1987 May;22(1):67–76. doi: 10.1002/jmv.1890220109. [DOI] [PubMed] [Google Scholar]
  20. Nielsen S. L., Sørensen I., Andersen H. K. Kinetics of specific immunoglobulins M, E, A, and G in congenital, primary, and secondary cytomegalovirus infection studied by antibody-capture enzyme-linked immunosorbent assay. J Clin Microbiol. 1988 Apr;26(4):654–661. doi: 10.1128/jcm.26.4.654-661.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Ripalti A., Boccuni M. C., Campanini F., Bergamini G., Lazzarotto T., Battista M. C., Dalla Casa B., Landini M. P. Construction of a polyepitope fusion antigen of human cytomegalovirus ppUL32 and detection of specific antibodies by ELISA. New Microbiol. 1995 Jan;18(1):1–12. [PubMed] [Google Scholar]
  22. Ripalti A., Dal Monte P., Boccuni M. C., Campanini F., Lazzarotto T., Campisi B., Ruan Q., Landini M. P. Prokaryotic expression of a large fragment of the most antigenic cytomegalovirus DNA-binding protein (ppUL44) and its reactivity with human antibodies. J Virol Methods. 1994 Jan;46(1):39–50. doi: 10.1016/0166-0934(94)90015-9. [DOI] [PubMed] [Google Scholar]
  23. Robinson J. M., Pilot-Matias T. J., Pratt S. D., Patel C. B., Bevirt T. S., Hunt J. C. Analysis of the humoral response to the flagellin protein of Borrelia burgdorferi: cloning of regions capable of differentiating Lyme disease from syphilis. J Clin Microbiol. 1993 Mar;31(3):629–635. doi: 10.1128/jcm.31.3.629-635.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Stagno S., Tinker M. K., Elrod C., Fuccillo D. A., Cloud G., O'Beirne A. J. Immunoglobulin M antibodies detected by enzyme-linked immunosorbent assay and radioimmunoassay in the diagnosis of cytomegalovirus infections in pregnant women and newborn infants. J Clin Microbiol. 1985 Jun;21(6):930–935. doi: 10.1128/jcm.21.6.930-935.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Vornhagen R., Hinderer W., Sonneborn H. H., Bein G., Matter L., The T. H., Jahn G., Plachter B. The DNA-binding protein pUL57 of human cytomegalovirus is a major target antigen for the immunoglobulin M antibody response during acute infection. J Clin Microbiol. 1995 Jul;33(7):1927–1930. doi: 10.1128/jcm.33.7.1927-1930.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Vornhagen R., Plachter B., Hinderer W., The T. H., Van Zanten J., Matter L., Schmidt C. A., Sonneborn H. H., Jahn G. Early serodiagnosis of acute human cytomegalovirus infection by enzyme-linked immunosorbent assay using recombinant antigens. J Clin Microbiol. 1994 Apr;32(4):981–986. doi: 10.1128/jcm.32.4.981-986.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. van Loon A. M., Heessen F. W., van der Logt J. T., van der Veen J. Direct enzyme-linked immunosorbent assay that uses peroxidase-labeled antigen for determination of immunoglobulin M antibody to cytomegalovirus. J Clin Microbiol. 1981 Mar;13(3):416–422. doi: 10.1128/jcm.13.3.416-422.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. van der Bij W., Torensma R., van Son W. J., Anema J., Schirm J., Tegzess A. M., The T. H. Rapid immunodiagnosis of active cytomegalovirus infection by monoclonal antibody staining of blood leucocytes. J Med Virol. 1988 Jun;25(2):179–188. doi: 10.1002/jmv.1890250208. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Clinical Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES