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. 1994 Apr;32(4):981–986. doi: 10.1128/jcm.32.4.981-986.1994

Early serodiagnosis of acute human cytomegalovirus infection by enzyme-linked immunosorbent assay using recombinant antigens.

R Vornhagen 1, B Plachter 1, W Hinderer 1, T H The 1, J Van Zanten 1, L Matter 1, C A Schmidt 1, H H Sonneborn 1, G Jahn 1
PMCID: PMC267166  PMID: 8027354

Abstract

DNA fragments from eight different reading frames of human cytomegalovirus (HCMV) were generated by PCR and subsequently cloned and expressed in Escherichia coli in fusion with glutathione S-transferase. The recombinant viral antigens were evaluated in immunoblot analyses. The most reactive antigens were purified and further evaluated in ELISAs. For this, sera from healthy blood donors and immunocompetent individuals with acute HCMV infection, and follow-up sera from transplant recipients with acute primary HCMV infection were used. The results of our experiments indicate that only three particular recombinant polypeptides from two viral proteins are necessary for serodiagnosis. While a fragment covering amino acids (aa) 495 to 691 of pp150 (150/1) was the most suitable antigen for the identification of infected individuals in general, immunoglobulin M antibodies against the C-terminal parts of pp150 (aa 862 to 1048; 150/7) and p52 (aa 297 to 433; 52/3) proved to be excellent serological markers to monitor acute HCMV infection. The selected recombinant antigens enable the improvement of serodiagnosis of HCMV-related diseases, especially during the early stages of infection.

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Selected References

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  1. Akrigg A., Wilkinson G. W., Oram J. D. The structure of the major immediate early gene of human cytomegalovirus strain AD169. Virus Res. 1985 Mar;2(2):107–121. doi: 10.1016/0168-1702(85)90242-4. [DOI] [PubMed] [Google Scholar]
  2. 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]
  3. Baur A., Vornhagen R., Korn K., Sonneborn H. H., Eberlein B., Harrer T., Brockhaus W., Jahn G. Viral culture and p24 antigenemia of human immunodeficiency virus (HIV)-infected individuals correlated with antibody profiles determined with recombinant polypeptides of all HIV-1 open-reading frames. J Infect Dis. 1992 Mar;165(3):419–426. doi: 10.1093/infdis/165.3.419. [DOI] [PubMed] [Google Scholar]
  4. Chang C. K., Balachandran N. Identification, characterization, and sequence analysis of a cDNA encoding a phosphoprotein of human herpesvirus 6. J Virol. 1991 Jun;65(6):2884–2894. doi: 10.1128/jvi.65.6.2884-2894.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. 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]
  6. Fleckenstein B., Müller I., Collins J. Cloning of the complete human cytomegalovirus genome in cosmids. Gene. 1982 Apr;18(1):39–46. doi: 10.1016/0378-1119(82)90054-3. [DOI] [PubMed] [Google Scholar]
  7. Galea G., Urbaniak S. J. Cytomegalovirus studies on blood donors in north-east Scotland and a review of UK data. Vox Sang. 1993;64(1):24–30. doi: 10.1111/j.1423-0410.1993.tb02510.x. [DOI] [PubMed] [Google Scholar]
  8. Gleaves C. A., Smith T. F., Shuster E. A., Pearson G. R. Comparison of standard tube and shell vial cell culture techniques for the detection of cytomegalovirus in clinical specimens. J Clin Microbiol. 1985 Feb;21(2):217–221. doi: 10.1128/jcm.21.2.217-221.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Gorgievski-Hrisoho M., Hinderer W., Nebel-Schickel H., Horn J., Vornhagen R., Sonneborn H. H., Wolf H., Siegl G. Serodiagnosis of infectious mononucleosis by using recombinant Epstein-Barr virus antigens and enzyme-linked immunosorbent assay technology. J Clin Microbiol. 1990 Oct;28(10):2305–2311. doi: 10.1128/jcm.28.10.2305-2311.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Kraat Y. J., Hendrix R. M., Landini M. P., Bruggeman C. A. Comparison of four techniques for detection of antibodies to cytomegalovirus. J Clin Microbiol. 1992 Feb;30(2):522–524. doi: 10.1128/jcm.30.2.522-524.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Landini M. P., Guan M. X., Jahn G., Lindenmaier W., Mach M., Ripalti A., Necker A., Lazzarotto T., Plachter B. Large-scale screening of human sera with cytomegalovirus recombinant antigens. J Clin Microbiol. 1990 Jun;28(6):1375–1379. doi: 10.1128/jcm.28.6.1375-1379.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Landini M. P., Lazzarotto T., Ertl P. F. Humoral immune response to human cytomegalovirus DNA polymerase. J Clin Microbiol. 1993 Mar;31(3):724–726. doi: 10.1128/jcm.31.3.724-726.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. 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]
  14. Landini M. P., Ripalti A., Sra K., Pouletty P. Human cytomegalovirus structural proteins: immune reaction against pp150 synthetic peptides. J Clin Microbiol. 1991 Sep;29(9):1868–1872. doi: 10.1128/jcm.29.9.1868-1872.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. 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]
  16. Lazzarotto T., Paya C. V., Smith T. F., Wiesner R. H., Krom R., Landini M. P. Antibody response to cytomegalovirus (CMV) polypeptides in liver transplant recipients with CMV hepatitis. Microbiologica. 1992 Jan;15(1):15–22. [PubMed] [Google Scholar]
  17. Lentz E. B., Dock N. L., McMahon C. A., Fiesthumel S. R., Arnold C. B., Lamberson H. V., Jr Detection of antibody to cytomegalovirus-induced early antigens and comparison with four serologic assays and presence of viruria in blood donors. J Clin Microbiol. 1988 Jan;26(1):133–135. doi: 10.1128/jcm.26.1.133-135.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Lindenmaier W., Necker A., Krause S., Bonewald R., Collins J. Cloning and characterization of major antigenic determinants of human cytomegalovirus Ad169 seen by the human immune system. Arch Virol. 1990;113(1-2):1–16. doi: 10.1007/BF01318349. [DOI] [PubMed] [Google Scholar]
  19. Plachter B., Klages S., Hagelmann S., Britt W., Landini M. P., Jahn G. Procaryotic expression of phosphorylated tegument protein pp65 of human cytomegalovirus and application of recombinant peptides for immunoblot analyses. J Clin Microbiol. 1990 Jun;28(6):1229–1235. doi: 10.1128/jcm.28.6.1229-1235.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Plachter B., Wieczorek L., Scholl B. C., Ziegelmaier R., Jahn G. Detection of cytomegalovirus antibodies by an enzyme-linked immunosorbent assay using recombinant polypeptides of the large phosphorylated tegument protein pp150. J Clin Microbiol. 1992 Jan;30(1):201–206. doi: 10.1128/jcm.30.1.201-206.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Schmidt C. A., Oettle H., Neuhaus P., Wiens M., Timm H., Wilborn F., Siegert W. Demonstration of cytomegalovirus by polymerase chain reaction after liver transplantation. Transplantation. 1993 Oct;56(4):872–874. doi: 10.1097/00007890-199310000-00019. [DOI] [PubMed] [Google Scholar]
  22. Smith D. B., Johnson K. S. Single-step purification of polypeptides expressed in Escherichia coli as fusions with glutathione S-transferase. Gene. 1988 Jul 15;67(1):31–40. doi: 10.1016/0378-1119(88)90005-4. [DOI] [PubMed] [Google Scholar]
  23. Sundqvist V. A., Xu W., Wahren B. Antibody responses to synthetic peptides from cytomegalovirus phosphoprotein 150. J Clin Microbiol. 1992 Oct;30(10):2735–2739. doi: 10.1128/jcm.30.10.2735-2739.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Van der Giessen M., van den Berg A. P., van der Bij W., Postma S., van Son W. J., The T. H. Quantitative measurement of cytomegalovirus-specific IgG and IgM antibodies in relation to cytomegalovirus antigenaemia and disease activity in kidney recipients with an active cytomegalovirus infection. Clin Exp Immunol. 1990 Apr;80(1):56–61. doi: 10.1111/j.1365-2249.1990.tb06441.x. [DOI] [PMC free article] [PubMed] [Google Scholar]

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