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.
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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
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
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
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
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
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
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
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
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
Higuchi, R. and Kwok, S.: Avoiding false positive with PCR.Nature 339: 237–238, 1989
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
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
Marsh, S. G. E. and Bodmer, J. G.: HLA-DR and DQ epitopes and monoclonal antibody specificity.Immunol Today 10: 305–312, 1989
Mullis, K. B. and Faloona, F.: Specific synthesis of DNA in vitro via polymerase catalyzed chain reaction.Methods Enzymol 155: 335–350, 1987
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
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
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
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
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
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
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
WHO Nomenclature Committee: Nomenclature for factors of theHLA system, 1989. Immunogenetics31: 131–140, 1990
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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
- Polymerase Chain Reaction
- Prime Pair
- Sequence Diversity
- Horseradish Peroxidase