Skip to main content
Log in

Rapid typing of HLA-DQB1 DNA polymorphism using nonradioactive oligonucleotide probes and amplified DNA

  • Original articles
  • Published:
Immunogenetics Aims and scope Submit manuscript

Abstract

The allelic sequence diversity at theHLA-DQBI locus has been analyzed by polymerase chain reaction (PCR) amplification and sequencing. Fifteen amino acid sequence-defined alleles (one previously unreported) and several silent nucleotide polymorphisms which subdivide these alleles have been identified. Here, we describe the specific amplification of theDQB1 second exon by several different PCR primer pairs and a simple and rapid typing procedure using a panel of 16 horseradish peroxidase (HRP)-labeled oligonucleotide probes capable of distinguishing theseDQBI alleles.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Angelini, G., de Preval, C., Gorski, J., and Mach, B.: High resolution analysis of the human HLA-DR polymorphism by hybridization with sequence-specific oligonuclemide probes.Proc Natl Acad Sci USA 83: 4489–4493, 1986

    Google Scholar 

  • Babbitt, B. P., Allen, P. M., Matsueda, G., Haber, E., and Unanue, E. R.: Binding of immunogenic peptides to la histocompatibility molecules.Nature 317: 359–361, 1985

    Google Scholar 

  • Bugawan, T. L., Saiki, R. K., Levenson, C. H., Watson, R. M., and Erlich, H. A.: The use of non-radioactive oligonucleotide probes to analyze enzymatically amplified DNA for prenatal diagnosis and forensic HLA typing.Bio/Technology 6: 943–947, 1988

    Google Scholar 

  • Bugawan, T. L., Begovich, A. B., and Erlich, H. A.: Rapid HLA-DPB typing using enzymatically amplified DNA and nonradioactive sequence-specific oligonucleotide probes.Immunogenetics 32: 231–241, 1990

    Google Scholar 

  • Buus, S., Sette, A., Colon, S., Miles, C., and Grey, H. M.: The relation between major histocompatibility complex (MHC) restriction and the capacity of la to bind immunogenic peptides.Science 235: 1353–1358, 1987

    Google Scholar 

  • Conner, B. J., Reyes, A. A., Morin, C., Itakura, K., Teplitz, R. L., and Wallace, R. B.: Detection of sickle cell β-S globin allele by hybridization with synthetic oligo nucleotides.Proc Natl Acad Sci USA 80: 278–282, 1983

    Google Scholar 

  • Erlich, H. A., Griffith, R., Bugawan, T. L., Ziegler, R., Alper, C., and Eisenbarth, G.: The identification of type 1 diabetic siblings with a novel HLA-DQB1 allele in genetic susceptibility and resistance.Diabetes, in press, 1990

  • Fronek, Z., Timmerman, L. A., Alper, C. A., Hahn, B. H., Kalunian, K., Peterlin, P. M., and McDevitt, H. O.: Major histocompatibility complex associations with systemic lupus erythematosus.Am J Med 85 (Suppl 6A): 42–44, 1988

    Google Scholar 

  • Guillet, J. G., Lai, M. Z., Briner, T. J., Buus, S., Sette, A., Grey, H. M., Smith, J. A., and Gefter, M. L.: Immunological self, nonself discrimination.Science 235: 865–870, 1987

    Google Scholar 

  • Gyllensten, U. B., Lashkari, D., and Erlich, H. A.: Allelic diversification at the HLA-DQβ locus of the mammalian major histocompatibility complex.Proc Natl Acad Sci USA 87: 1835–1839, 1990

    Google Scholar 

  • Higuchi, R. and Kwok, S.: Avoiding false positive with PCR.Nature 339: 237–238, 1989

    Google Scholar 

  • Horn, G. T., Bugawan, T. L., Long, C., and Erlich, H. A.: Allelic sequence variation of the HLA-DQ loci: relationship to serology and insulin-dependent diabetes susceptibility.Proc Natl Acad Sci USA 85: 6012–6016, 1988

    Google Scholar 

  • Levenson, C. H. and Chang, C. A.: Non isotopically labelled probes and primers. In M. Innis, J. Sninsky, D. Gelfand, and T. White (eds.):PCR Protocols and Application -A Laboratory Manual, p. 99–112, Academic, New York, 1986

    Google Scholar 

  • Marsh, S. G. E. and Bodmer, J. G.: HLA-DR and DQ epitopes and monoclonal antibody specificity.Immunol Today 10: 305–312, 1989

    Google Scholar 

  • Mullis, K. B. and Faloona, F.: Specific synthesis of DNA in vitro via polymerase catalyzed chain reaction.Methods Enzymol 155: 335–350, 1987

    Google Scholar 

  • Saiki, R. K., Scharf, S., Faloona, F., Mullis, K. B., Horn, G. T., Erlich, H. A., and Arnheim, N.: Enzymatic amplification ofβ-globin genomic sequence and restriction site analysis for diagnosis of sickle cell anemia.Science 230: 1350–1354, 1985

    Google Scholar 

  • Saiki, R. K., Bugawan, T. L., Horn, G. T., Mullis, K. B., and Erlich, H. A.: Analysis of enzymatically amplifiedβ-globin and HLADQα DNA with allele specific probes.Nature 324: 163–166, 1986

    Google Scholar 

  • Saiki, R. K., Gelfand, D. H., Stoffel, S., Scharf, S. J., Higuchi, R., Horn, G. T., Mullis, K. B., and Erlich, H. A.: Primer-directed enzymatic amplification of DNA with thermostable DNA polymerase.Science 239: 487–491, 1988

    Google Scholar 

  • Saiki, R., Walsh, P. S., Levenson, C. H., and Erlich, H. A.: Genetic analysis of amplified DNA with immobilized sequence-specific oligonucleotide probes.Proc Natl Acad Sci USA 86: 6230–6234, 1989

    Google Scholar 

  • Scharf, S. J., Friedmann, A., Brautbar, C., Szafer, F., Steinman, L., Horn, G., Gyllensten, U., and Erlich, H. A.: HLA class II allelic variation and susceptibility to Pemphigus vulgaris.Proc Natl Acad Sci USA 85: 3504–3508, 1988

    Google Scholar 

  • Scharf, S., Friedmann, A., Steinman, L., Brautbar, C., and Erlich, H. A.: Specific HLA-DQβ and DROI alleles confer susceptibility to Pemphigus vulgaris.Proc Natl Acad Sci USA 86: 6215–6219, 1989

    Google Scholar 

  • Sette, A., Buns, S., Colon, S., Smith, J. A., Miles, C., and Grey, H. M.: Structural characteristics of an antigen required for its interaction with Ia and recognition by T cells.Nature 328: 395–399, 1987

    Google Scholar 

  • WHO Nomenclature Committee: Nomenclature for factors of theHLA system, 1989. Immunogenetics31: 131–140, 1990

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Bugawan, T.L., Erlich, H.A. Rapid typing of HLA-DQB1 DNA polymorphism using nonradioactive oligonucleotide probes and amplified DNA. Immunogenetics 33, 163–170 (1991). https://doi.org/10.1007/BF01719235

Download citation

  • Received:

  • Revised:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01719235

Keywords

Navigation