Skip to main content
Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1993 Dec;31(12):3157–3164. doi: 10.1128/jcm.31.12.3157-3164.1993

Detection of immunoglobulin M antibodies to glycoprotein G-2 by western blot (immunoblot) for diagnosis of initial herpes simplex virus type 2 genital infections.

D W Ho 1, P R Field 1, W L Irving 1, D R Packham 1, A L Cunningham 1
PMCID: PMC266368  PMID: 7508453

Abstract

Western blots (immunoblots) for the detection of immunoglobulin M (IgM) antibodies specific for herpes simplex virus type 1 (HSV-1) and HSV-2 in patients' sera were developed. The locations of the type-specific glycoprotein G (gpG-2) of HSV-2 (92- and 140-kDa forms) and glycoprotein C of HSV-1 (gpC-1), which carries mostly type-specific antigenic epitopes, were checked with specific monoclonal antibodies. Western blot assays for IgM antibody to gpC-1 or gpG-2 were performed after depletion of IgG by precipitation with anti-human IgG. In patients with primary HSV-2 genital infections, seroconversion of IgM and IgG antibodies to both the 92- and 140-kDa forms of gpG-2 was observed, although both antibodies appeared in convalescent-phase serum after the first week. IgM and IgG antibodies to low-molecular-size polypeptides (40 to 65 kDa) were the first antibodies observed in patients with primary infection, but these antibodies were cross-reactive with HSV-1 and HSV-2. However, in patients with recurrent HSV-2 infections, IgG antibodies to both forms of gpG-2 and the low-molecular-size polypeptides were found no matter how early after onset the patient was bled, and IgM to gpG-2 did not appear. In patients with nonprimary initial genital HSV-2 infections, IgG antibody to HSV-1 was demonstrated in the first serum specimen, and HSV-2-specific IgM was found in 39% of the serum specimens. Hence, the Western blot assay can be used to test for IgM antibody to gpG-2, allowing for the retrospective diagnosis of inital HSV-2 infections and its use as a supplementary test to the gpG-2 IgG enzyme-linked immunosorbent assays developed elsewhere. In contrast, IgM antibody to gpG-2 is not usually detected in patients with recurrent HSV-2 infections.

Full text

PDF
3157

Images in this article

Selected References

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

  1. Arvin A. M., Koropchak C. M., Yeager A. S., Pereira L. Detection of type-specific antibody to herpes simplex virus type 1 by radioimmunoassay with herpes simplex virus type 1 glycoprotein C purified with monoclonal antibody. Infect Immun. 1983 Apr;40(1):184–189. doi: 10.1128/iai.40.1.184-189.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Ashley R. L. Genital herpes infections. Clin Lab Med. 1989 Sep;9(3):405–420. [PubMed] [Google Scholar]
  3. Ashley R. L., Militoni J., Lee F., Nahmias A., Corey L. Comparison of Western blot (immunoblot) and glycoprotein G-specific immunodot enzyme assay for detecting antibodies to herpes simplex virus types 1 and 2 in human sera. J Clin Microbiol. 1988 Apr;26(4):662–667. doi: 10.1128/jcm.26.4.662-667.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Ashley R. L., Militoni J. Use of densitometric analysis for interpreting HSV serologies based on Western blot. J Virol Methods. 1987 Nov;18(2-3):159–168. doi: 10.1016/0166-0934(87)90121-2. [DOI] [PubMed] [Google Scholar]
  5. Ashley R., Benedetti J., Corey L. Humoral immune response to HSV-1 and HSV-2 viral proteins in patients with primary genital herpes. J Med Virol. 1985 Oct;17(2):153–166. doi: 10.1002/jmv.1890170208. [DOI] [PubMed] [Google Scholar]
  6. Ashley R., Cent A., Maggs V., Nahmias A., Corey L. Inability of enzyme immunoassays to discriminate between infections with herpes simplex virus types 1 and 2. Ann Intern Med. 1991 Oct 1;115(7):520–526. doi: 10.7326/0003-4819-115-7-520. [DOI] [PubMed] [Google Scholar]
  7. Bernstein D. I., Bryson Y. J., Lovett M. A. Antibody response to type-common and type-unique epitopes of herpes simplex virus polypeptides. J Med Virol. 1985 Mar;15(3):251–263. doi: 10.1002/jmv.1890150306. [DOI] [PubMed] [Google Scholar]
  8. Bernstein D. I. Effects of prior HSV-1 infection on genital HSV-2 infection. Prog Med Virol. 1991;38:109–127. [PubMed] [Google Scholar]
  9. Bernstein D. I., Lovett M. A., Bryson Y. J. Serologic analysis of first-episode nonprimary genital herpes simplex virus infection. Presence of type 2 antibody in acute serum samples. Am J Med. 1984 Dec;77(6):1055–1060. doi: 10.1016/0002-9343(84)90188-8. [DOI] [PubMed] [Google Scholar]
  10. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  11. Corey L., Adams H. G., Brown Z. A., Holmes K. K. Genital herpes simplex virus infections: clinical manifestations, course, and complications. Ann Intern Med. 1983 Jun;98(6):958–972. doi: 10.7326/0003-4819-98-6-958. [DOI] [PubMed] [Google Scholar]
  12. Eberle R., Courtney R. J. Assay of type-specific and type-common antibodies to herpes simplex virus types 1 and 2 in human sera. Infect Immun. 1981 Mar;31(3):1062–1070. doi: 10.1128/iai.31.3.1062-1070.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Eberle R., Mou S. W., Zaia J. A. Polypeptide specificity of the early antibody response following primary and recurrent genital herpes simplex virus type 2 infections. J Gen Virol. 1984 Oct;65(Pt 10):1839–1843. doi: 10.1099/0022-1317-65-10-1839. [DOI] [PubMed] [Google Scholar]
  14. Field P. R., Ho D. W., Irving W. L., Isaacs D., Cunningham A. L. The reliability of serological tests for the diagnosis of genital herpes: a critique. Pathology. 1993 Apr;25(2):175–179. doi: 10.3109/00313029309084794. [DOI] [PubMed] [Google Scholar]
  15. Gibbs R. S., Mead P. B. Preventing neonatal herpes--current strategies. N Engl J Med. 1992 Apr 2;326(14):946–947. doi: 10.1056/NEJM199204023261409. [DOI] [PubMed] [Google Scholar]
  16. Goldstein L. C., Corey L., McDougall J. K., Tolentino E., Nowinski R. C. Monoclonal antibodies to herpes simplex viruses: use in antigenic typing and rapid diagnosis. J Infect Dis. 1983 May;147(5):829–837. doi: 10.1093/infdis/147.5.829. [DOI] [PubMed] [Google Scholar]
  17. Ho D. W., Field P. R., Cunningham A. L. Rapid diagnosis of acute Epstein-Barr virus infection by an indirect enzyme-linked immunosorbent assay for specific immunoglobulin M (IgM) antibody without rheumatoid factor and specific IgG interference. J Clin Microbiol. 1989 May;27(5):952–958. doi: 10.1128/jcm.27.5.952-958.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Ho D. W., Field P. R., Sjögren-Jansson E., Jeansson S., Cunningham A. L. Indirect ELISA for the detection of HSV-2 specific IgG and IgM antibodies with glycoprotein G (gG-2). J Virol Methods. 1992 Mar;36(3):249–264. doi: 10.1016/0166-0934(92)90056-j. [DOI] [PubMed] [Google Scholar]
  19. Johnson R. E., Nahmias A. J., Magder L. S., Lee F. K., Brooks C. A., Snowden C. B. A seroepidemiologic survey of the prevalence of herpes simplex virus type 2 infection in the United States. N Engl J Med. 1989 Jul 6;321(1):7–12. doi: 10.1056/NEJM198907063210102. [DOI] [PubMed] [Google Scholar]
  20. Juto P., Settergren B. Specific serum IgA, IgG and IgM antibody determination by a modified indirect ELISA-technique in primary and recurrent herpes simplex virus infection. J Virol Methods. 1988 May;20(1):45–55. doi: 10.1016/0166-0934(88)90039-0. [DOI] [PubMed] [Google Scholar]
  21. Kahlon J., Whitley R. J. Antibody response of the newborn after herpes simplex virus infection. J Infect Dis. 1988 Nov;158(5):925–933. doi: 10.1093/infdis/158.5.925. [DOI] [PubMed] [Google Scholar]
  22. Kalimo K. O., Marttila R. J., Granfors K., Viljanen M. K. Solid-phase radioimmunoassay of human immunoglobulin M and immunoglobulin G antibodies against herpes simplex virus type 1 capsid, envelope, and excreted antigens. Infect Immun. 1977 Mar;15(3):883–889. doi: 10.1128/iai.15.3.883-889.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Kimmel N., Friedman M. G., Sarov I. Enzyme-linked immunosorbent assay (ELISA) for detection of herpes simplex virus-specific IgM antibodies. J Virol Methods. 1982 May;4(4-5):219–227. doi: 10.1016/0166-0934(82)90068-4. [DOI] [PubMed] [Google Scholar]
  24. Kulhanjian J. A., Soroush V., Au D. S., Bronzan R. N., Yasukawa L. L., Weylman L. E., Arvin A. M., Prober C. G. Identification of women at unsuspected risk of primary infection with herpes simplex virus type 2 during pregnancy. N Engl J Med. 1992 Apr 2;326(14):916–920. doi: 10.1056/NEJM199204023261403. [DOI] [PubMed] [Google Scholar]
  25. Kurtz J. B. Specific IgG and IgM antibody responses in herpes-simplex-virus infections. J Med Microbiol. 1974 Aug;7(3):333–341. doi: 10.1099/00222615-7-3-333. [DOI] [PubMed] [Google Scholar]
  26. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  27. Langenberg A., Benedetti J., Jenkins J., Ashley R., Winter C., Corey L. Development of clinically recognizable genital lesions among women previously identified as having "asymptomatic" herpes simplex virus type 2 infection. Ann Intern Med. 1989 Jun 1;110(11):882–887. doi: 10.7326/0003-4819-110-11-882. [DOI] [PubMed] [Google Scholar]
  28. Lee F. K., Coleman R. M., Pereira L., Bailey P. D., Tatsuno M., Nahmias A. J. Detection of herpes simplex virus type 2-specific antibody with glycoprotein G. J Clin Microbiol. 1985 Oct;22(4):641–644. doi: 10.1128/jcm.22.4.641-644.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Lee F. K., Pereira L., Griffin C., Reid E., Nahmias A. A novel glycoprotein for detection of herpes simplex virus type 1-specific antibodies. J Virol Methods. 1986 Sep;14(2):111–118. doi: 10.1016/0166-0934(86)90041-8. [DOI] [PubMed] [Google Scholar]
  30. Marsden H. S., Buckmaster A., Palfreyman J. W., Hope R. G., Minson A. C. Characterization of the 92,000-dalton glycoprotein induced by herpes simplex virus type 2. J Virol. 1984 May;50(2):547–554. doi: 10.1128/jvi.50.2.547-554.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. McClung H., Seth P., Rawls W. E. Relative concentrations in human sera of antibodies to cross-reacting and specific antigens of Herpes simplex virus types 1 and 2. Am J Epidemiol. 1976 Aug;104(2):192–201. doi: 10.1093/oxfordjournals.aje.a112289. [DOI] [PubMed] [Google Scholar]
  32. Nowinski R. C., Tam M. R., Goldstein L. C., Stong L., Kuo C. C., Corey L., Stamm W. E., Handsfield H. H., Knapp J. S., Holmes K. K. Monoclonal antibodies for diagnosis of infectious diseases in humans. Science. 1983 Feb 11;219(4585):637–644. doi: 10.1126/science.6297006. [DOI] [PubMed] [Google Scholar]
  33. Olofsson S., Lundström M., Marsden H., Jeansson S., Vahlne A. Characterization of a herpes simplex virus type 2-specified glycoprotein with affinity for N-acetylgalactosamine-specific lectins and its identification as g92K or gG. J Gen Virol. 1986 Apr;67(Pt 4):737–744. doi: 10.1099/0022-1317-67-4-737. [DOI] [PubMed] [Google Scholar]
  34. Ozaki Y., Ishiguro T. Relationship between type specific antibodies to herpes simplex virus and type specificity in human sera. J Med Virol. 1983;12(2):103–109. doi: 10.1002/jmv.1890120204. [DOI] [PubMed] [Google Scholar]
  35. Para M. F., Zezulak K. M., Conley A. J., Weinberger M., Snitzer K., Spear P. G. Use of monoclonal antibodies against two 75,000-molecular-weight glycoproteins specified by herpes simplex virus type 2 in glycoprotein identification and gene mapping. J Virol. 1983 Mar;45(3):1223–1227. doi: 10.1128/jvi.45.3.1223-1227.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Plummer G. A review of the identification and titration of antibodies to herpes simplex viruses type 1 and type 2 in human sera. Cancer Res. 1973 Jun;33(6):1469–1476. [PubMed] [Google Scholar]
  37. Prober C. G., Arvin A. M. Genital herpes and the pregnant woman. Curr Clin Top Infect Dis. 1989;10:1–26. [PubMed] [Google Scholar]
  38. Roizman B., Norrild B., Chan C., Pereira L. Identification and preliminary mapping with monoclonal antibodies of a herpes simplex virus 2 glycoprotein lacking a known type 1 counterpart. Virology. 1984 Feb;133(1):242–247. doi: 10.1016/0042-6822(84)90447-1. [DOI] [PubMed] [Google Scholar]
  39. Sjöblom I., Lundström M., Sjögren-Jansson E., Glorioso J. C., Jeansson S., Olofsson S. Demonstration and mapping of highly carbohydrate-dependent epitopes in the herpes simplex virus type 1-specified glycoprotein C. J Gen Virol. 1987 Feb;68(Pt 2):545–554. doi: 10.1099/0022-1317-68-2-545. [DOI] [PubMed] [Google Scholar]
  40. Suchánková A., Hirsch I., Kremár M., Vonka V. Determination of herpes simplex virus type-specific antibodies by solid-phase RIA on Helix pomatia lectin-purified antigens. J Infect Dis. 1984 Jun;149(6):964–972. doi: 10.1093/infdis/149.6.964. [DOI] [PubMed] [Google Scholar]
  41. Vestergaard B. F. Herpes simplex virus antigens and antibodies: a survey of studies based on quantitative immunoelectrophoresis. Rev Infect Dis. 1980 Nov-Dec;2(6):899–913. doi: 10.1093/clinids/2.6.899. [DOI] [PubMed] [Google Scholar]
  42. Zezulak K. M., Spear P. G. Characterization of a herpes simplex virus type 2 75,000-molecular-weight glycoprotein antigenically related to herpes simplex virus type 1 glycoprotein C. J Virol. 1983 Sep;47(3):553–562. doi: 10.1128/jvi.47.3.553-562.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Zezulak K. M., Spear P. G. Mapping of the structural gene for the herpes simplex virus type 2 counterpart of herpes simplex virus type 1 glycoprotein C and identification of a type 2 mutant which does not express this glycoprotein. J Virol. 1984 Mar;49(3):741–747. doi: 10.1128/jvi.49.3.741-747.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Zweig M., Heilman C. J., Jr, Bladen S. V., Showalter S. D., Hampar B. Detection in antisera of antibodies that cross-react with herpes simplex virus type 1 glycoprotein gC. Infect Immun. 1983 Aug;41(2):482–487. doi: 10.1128/iai.41.2.482-487.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Zweig M., Showalter S. D., Simms D. J., Hampar B. Antibodies to a synthetic oligopeptide that react with herpes simplex virus type 1 and 2 glycoprotein C. J Virol. 1984 Aug;51(2):430–436. doi: 10.1128/jvi.51.2.430-436.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES